HK1174301B - Sample plate - Google Patents

Description

Sample plate

Technical Field

The present invention relates to a sample plate, an automated device, a reagent bead or microsphere dispenser, a method of dispensing reagent beads or microspheres, a kit for performing an enzyme linked immunosorbent assay procedure, a kit for performing a nucleic acid detection procedure, a method of manufacturing a sample plate and a computer program executable by a control system of an automated device.

Preferred embodiments relate to an automated reagent bead or microsphere dispenser for dispensing reagent beads or microspheres into a sample plate. The sample plate may be used to perform a diagnostic assay, such as an enzyme-linked immunosorbent assay (ELISA) procedure or other immunoassay procedure. Alternatively, the sample plate may be used to perform testing of DNA or RNA sequences.

Background

Immunoassay procedures are the preferred means of testing biological products. These processes take advantage of the ability of body-produced antibodies to recognize and combine with specific antigens (e.g., that can be associated with foreign objects such as bacteria or viruses, or with other body products such as hormones). Once a particular antigen-antibody combination has occurred, detection can be by using chromogens, fluorescent or chemiluminescent materials or less preferably by using radioactive substances. Radioactive materials are less preferred because of environmental and safety issues associated with their handling, storage, and disposal. The same principle can be used to detect or identify any material that forms a specific binding pair, for example using lectins, rheumatoid factors, protein a or nucleic acids as one of the binding partners.

ELISA is a particularly preferred form of immunoassay process, wherein one element of a binding pair is attached to an insoluble support surface ("solid phase") such as a sample container, and the binding pair is detected after the reaction ("conjugated") by using yet another specific binding agent conjugated to an enzyme. The process of ELISA is well known in the art and has been used for many years for research and commercial purposes. Many books and review articles describe the theory and practice of immunoassays. For example, suggestions are given for the properties and selection of the solid phase of the capture assay, for methods and reagents for coating the solid phase with capture components, for the properties and selection of labels, and for methods of labeling components. An example of a standard textbook is "ELISA and Other solid phase Immunoassays, the Theoretical and Practical Aspects of ELISA and Other solid phase Immunoassays", John Wiley, Editory D.M. Kemeny & S.J. Challacomb, published 1988. These suggestions are also applicable to the determination of other specific binding pairs.

In the most common type of ELISA, the solid phase is covered with one of the binding pairs. An aliquot to be examined is incubated with a solid-coated solid phase and any analyte that may be present is captured on the solid phase. After washing to remove any residual sample and any interfering materials that it may contain, a second binding agent, specific for the analyte and conjugated to an enzyme, is added to the solid phase. During the second incubation, any analyte captured on the solid phase will bind to the conjugate. After a second wash to remove any unbound conjugate, a color-developing substrate for the enzyme is added to the solid phase. Any enzyme present will begin to convert the substrate into a chromonic product. After a certain time, the amount of product formed is measured with a spectrophotometer, either directly or after stopping the reaction.

It will be appreciated that the above is only a rough description of the overall process of the bioassay and that many variations are known in the art, including fluorescent and luminescent substrates for ELISA, direct labeling of the second element of the binding pair with fluorescent or luminescent molecules (in which case the process is no longer referred to as ELISA, but the processing steps are very similar) and nucleic acids or other specific pairing agents instead of antibodies as binders. However, all assays require that a fluid sample (e.g., blood, serum, urine, etc.) be aspirated from the sample tube and then dispensed into the solid phase. The samples may be diluted prior to dispensing into the solid phase or they may be dispensed into a deep well microplate, diluted there and the diluted analyte then transferred to the functional solid phase.

The most common type of solid phase is a standard sample container known as a microplate, which can be easily stored and used with many biological samples. Microplates have been commercially available since the 60's of the 20 th century and are made of, for example, polystyrene, PVC, Perspex or Lucite and are approximately 5 inches (12.7 cm) in length, 3.3 inches (8.5 cm) in width and 0.55 inches (1.4 cm) in depth. Microplates made of polystyrene are particularly preferred because polystyrene improves the optical clarity that aids in the visual observation of any reaction results. Polystyrene microplates are also compact, lightweight, and easy to clean. Microplates manufactured by the applicant are sold under the name "MICROTITRE" (RTM). Known microplates comprise 96 wells (also commonly referred to as "microwells") arranged symmetrically in an 8 x 12 array. Microwells typically have a maximum capacity of about 350 μ l. However, typically only 10-200. mu.l of fluid is dispensed into the microwell. In some arrangements of microplates, the microwells may be arranged in strips of 8 or 12 wells, which can be moved or combined in a carrier to form a complete plate of conventional size.

Forward and reverse control are typically supplied in commercial packages and are used for quality control and to provide relative trade-offs. After reading the processed microplate, the control results are checked against the manufacturer's confirmation values to ensure that the analysis has operated correctly and then the values are used to distinguish the forward sample from the reverse sample and calculate the cut-off values. Standards are generally provided for quantitative determinations and are used to construct a standard curve whereby the concentration of an analyte in a sample can be interpolated.

It will be appreciated that the ELISA process as outlined above involves a number of steps including pipetting, incubation, washing, transporting microplates between activities, reading and data analysis. In recent years, systems have been developed that automate the steps involved in ELISA procedures such as sample dispensing, dilution, incubation at specific temperatures, washing, enzyme conjugate addition, reagent addition, reaction stop, and result analysis. The pipetting mechanism used to aspirate and dispense the fluid sample uses disposable tips that are ejected after use to prevent cross-contamination of the patient sample. Multiple instrument controls are in place to ensure proper volume, times, wavelength and temperature are employed, and data transfer and analysis is fully verified and monitored. Automated immunoassay devices for performing ELISA processes are now widely used, for example, in laboratories of pharmaceutical companies, veterinary and plant laboratories, hospitals and universities, for in vitro diagnostic applications such as disease and infection testing, and for assisting in the production of new vaccines and pharmaceuticals.

ELISA kits are commercially available, consisting of a microplate with microwells that have been coated by manufacturers with specific antibodies (or antigens). For example, in the case of a hepatitis B antigen diagnostic kit, the manufacturer of the kit dispenses anti-hepatitis B antibodies, which have been suspended in a fluid, into the microwells of a microplate. The microplate is then incubated for a period of time during which the antibodies adhere to the walls of the microwells up to the liquid fill level (typically about half the maximum fluid capacity of the microwell). The microwells are then washed, leaving the microplate with the walls of the microwells uniformly coated with anti-hepatitis B antibody to the liquid fill level.

The test laboratory will receive a number of sample tubes containing e.g. body fluids from a number of patients. A specified amount of fluid is then aspirated from the sample tube using a pipetting mechanism and then dispensed into one or more microwells of a microplate that has been previously prepared by the manufacturer as described above. If it is desired to test a patient for several different diseases, fluid from the patient must be dispensed into several separate microplates, each coated with a different adhesive by its manufacturer. Each microplate can then be processed separately to detect the presence or absence of a different disease. It will be seen that multiple microplates are required to analyse several different analytes and that aliquots of the same sample are transferred to different microplates. This results in a large number of processing steps and incubators and washing stations that can process many microplates virtually simultaneously. In automated systems, this requires instruments with multiple incubators and complex programming to avoid conflicts in different requirements between microplates. For manual operation, either several full-time staff are required or throughput of samples is slow. It is possible to combine the combined strips of differently coated microwells into a single carrier, add a single sample equally to the different types of wells and then perform an ELISA in this combined microwell. However, limitations on assay development make this combination difficult to achieve and it is known in the art that users combining strips in this manner can lead to errors in result assignment, while the manufacture of microplates with several different coating layers in different microwells also presents quality control difficulties.

Conventional ELISA techniques have focused on performing the same single test on multiple patient samples per microplate or detecting the presence or absence of one or more of these patients in multiplex analytes without distinguishing between the actual presence of possible analytes. For example, it is common to determine whether a patient has antibodies to HIV-1 or HIV-2, or antigens of HIV-1 or HIV-2, in a single microwell, without determining which analyte is present, and similarly for HCV antibodies and antigens.

However, a new generation of assays is being developed that enable multiplexed execution. Multiplexing enables multiple different tests to be performed simultaneously on the same patient sample.

One recent multiplexing method is to provide a microplate comprising 96 sample microwells, wherein an array of different capture antibodies is disposed in each sample microwell. The array comprises an array of 20nl spots each having a diameter of 350 μm. The dots are arranged at a pitch interval of 650 μm. Each spot corresponds to a different capture antibody.

Multiplexing allows for more data points and more information per assay than with conventional ELISA techniques, where each sample plate tests for a single analyte of interest. The ability to combine multiple separate tests into the same assay can result in considerable time and cost savings. Multiplexing also enables a reduction in the overall footprint of the automation device.

Despite the many advantages of the presently known ELISA techniques and the new multiplexing techniques currently under development, it is still desirable to provide a sample plate and associated automated apparatus with an improved format and offering greater flexibility than existing ELISA arrangements.

In addition to ELISA procedures, it is also known to use hybridization probes to test for the presence of DNA or RNA sequences. Hybridization probes generally include DNA or RNA fragments that are used to detect the presence of a nucleotide sequence complementary to the DNA or RNA sequence on the probe. Hybridization probes hybridize, due to their complementarity to the sample being analyzed, to single-stranded nucleic acids (e.g., DNA or RNA) whose base sequence allows pairing. The hybridization probes may be labeled or tagged with a molecular label, such as a radioactive or more preferably fluorescent molecule. The probe is inert until a point of hybridization at which a conformational change occurs, and molecular synthesis begins to move and then fluorescent (detectable under ultraviolet light) DNA sequences or RNA transcripts with moderate to high sequence similarity to the probe are detected by observing the probe under ultraviolet light.

An assay device and assembly for detecting an analyte in a liquid sample is disclosed in US-5620853(Chiron Corporation). The assay device includes a molded well that includes fingers that project upward from the bottom of the well and into which reagent beads are dispensed. The reagent beads are captured in the fingers but can still move up and down within the height of the fingers. The assay device is arranged to expose the reagent beads to as much fluid flow as possible and to rely on signals from the underside of the reagent beads to produce a result.

The arrangement disclosed in US-5620853 has several problems.

First, since the reagent beads can move up and down freely within the finger height, the reagent beads may get stuck at an undesired height during the processing or reading step. In particular, the well design is relatively delicate and complex and any movement of or damage to the fingers can cause the reagent beads to become stuck at an undesirable height. The fingers also protrude from the base, which makes them susceptible to damage, especially during the pipetting and rinsing stages. If the reagent beads do get stuck at an undesirable height within the fingers, there is likely to be a negative impact on the accuracy of the testing process.

Secondly, the design of the well in which the fingers are arranged to receive a single reagent bead is such that fluid is wicked in close proximity to the reagent bead and the reagent bead is covered by the fluid raised in the well. A single well requires approximately 300 mul of fluid. US-5620853 also discloses an arrangement in which a plurality of wells are in fluid communication with each other. For a multi-well procedure, approximately 300 μ l of fluid would be required for each well. It is therefore apparent that a multi-well arrangement requires an excessive amount of fluid to be dispensed relative to conventional systems.

Third, for a given size sample plate, the arrangement of the fingers reduces the maximum package seal of the wells so that relatively few tests can be performed for a given sample plate.

Fourth, the multi-well arrangement disclosed in US-5620853 is particularly susceptible to cross-talk.

Fifth, the arrangement disclosed in US-5620853 is such that, when a single bead is used, then the uniformity of the fluid is only affected by the protruding fingers. The well is likely to have areas where unmixed fluids will be captured. Multi-well arrangements also present a serious problem requiring that any fluid that passes over all the beads must traverse a tortuous path to pass from one well to another. This will cause serious problems with respect to fluid mixing and bead-to-bead repeatability. This single well arrangement is quite different from the in-line multi-well arrangement disclosed in US-5620853 and therefore the two different arrangements will have quite different fluid characteristics. This is likely to result in different fluid behavior depending on the arrangement used and therefore likely to vary significantly in the results depending on whether a single well or multiple wells are used. Although theoretically these two different arrangements could be independently identified, this would result in increased cost and reduced throughput.

Finally, the sample wells disclosed in US-5620853 are relatively complex to manufacture and are likely to suffer from unreliability issues during manufacture. Long and thin fingers are difficult to form by molding and will be prone to damage during manufacture or during use. The fingers also have a structure at the top which will be an undercut in the molding tool. When the part is ejected from the tool, the fingers must flex to allow this structure to pass over the tool material. Such manufacturing processes are generally undesirable due to unreliability issues. Moreover, any variation in process parameters is likely to affect the ability to release the part from the tool and cause the part to have incorrect mechanical tolerances. The position of the fingers relative to each other will be critical to allow the reagent beads to move up and down correctly and also to ensure that the reagent beads do not come out of the top of the fingers. In practice, this is very difficult to control in a mass production environment. It is also noted that the design of a single bead arrangement is quite different from the design of a multi-well arrangement. Thus, a completely different tool design would be required, which greatly increases manufacturing complexity. In a high volume manufacturing environment, the combination of design features and quality assurance issues will make the production of sample plates prohibitively expensive.

It is therefore desirable to provide an improved sample plate for holding reagent beads.

Disclosure of Invention

According to one aspect of the present invention there is provided a sample plate comprising one or more sample wells, wherein the one or more sample wells comprise a base and one or more pockets or recesses provided in the base, wherein the one or more pockets or recesses comprise a bore having a tapered section, wherein in use a reagent bead or microsphere is substantially retained or secured within the bore by the tapered section.

The bore with the tapered section should not be misinterpreted as, for example, a shallow or small depression in which the reagent bead or microsphere simply can reside but in which the reagent bead or microsphere is not substantially retained or secured.

The sample plate according to the invention is particularly advantageous compared to the sample plate disclosed in US-5620853.

According to a preferred embodiment, in use, the reagent bead or microsphere is substantially retained or secured within the bore by an interference or friction fit with the tapered section of the bore.

According to a preferred embodiment of the invention, the reagent beads are preferably inserted into a sample plate having a plurality of tapered holes or sections which serve to securely fasten or lock the reagent beads in place as they are inserted. The preset force is preferably used to insert the reagent beads. The preset force is preferably sufficient to compress the reagent bead and/or deform the tapered section of the bore so as to form or enhance an interference or friction fit with the tapered section of the bore.

The sample plate according to the invention is thus particularly robust during manufacture and in subsequent processing stages, including the stage of inserting reagent beads into the tapered holes and subsequent handling and processing of the sample plate. Once the reagent beads have been inserted into the sample plate, they are preferably not free to move in any direction and become substantially a fixed part of the sample plate. The angle of the taper is preferably arranged so that the reagent beads are locked or otherwise securely fastened in the holes, making the arrangement very reliable.

According to a preferred embodiment, in use, a reagent bead is preferably substantially retained or secured within a bore if the sample plate (i.e. the plane of the sample plate) is tilted more than 10 °, 20 °, 30 °, 40 °, 50 °, 60 °, 70 °, 80 ° or 90 ° relative to horizontal, or inverted.

According to a preferred embodiment, the opening to the aperture and/or the cross-sectional shape of the aperture (i.e. at a position intermediate the opening to the aperture and the base of the aperture) is circular. However, in less preferred embodiments, the cross-sectional shape of the opening and/or aperture may be substantially circular, elliptical, oblong, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, decagonal, or polygonal.

According to a preferred embodiment, the diameter of the opening of the aperture is preferably selected from the group consisting of: (i) less than 0.5 mm; (ii)0.5-1.0 mm; (iii)1.0-1.5 mm; (iv)1.5-2.0 mm; (v)2.0-2.5 mm; (vi)2.5-3.0 mm; (vii)3.0-3.5 mm; (viii)3.5-4.0 mm; (ix)4.0-4.5 mm; (x)4.5-5.0 mm; (xi) Less than 5.0 mm; and (xii) > 5.0 mm.

According to a preferred embodiment, the diameter of the opening of the bore is preferably larger than the diameter of the reagent bead or microsphere. If the opening of the bore has a cross-sectional shape other than circular, the smallest span of the cross-sectional shape of the bore at the opening is preferably larger than the diameter of the reagent bead or microsphere.

According to a preferred embodiment, the diameter of the bore, preferably at a position intermediate the opening of the bore and the base of the bore, is preferably at least 5% smaller than the diameter of the reagent bead or microsphere and/or preferably at least 5% smaller than the diameter of the opening of the bore. If the opening of the bore has a cross-sectional shape other than circular, the smallest span of the cross-sectional shape of the bore, preferably at a position intermediate the opening of the bore and the base of the bore, is preferably at least 5% smaller than the diameter of the reagent bead or microsphere and/or preferably at least 5% smaller than the diameter of the opening of the bore.

According to a preferred embodiment, the diameter of the bore, preferably at a position intermediate the opening of the bore and the base of the bore, is preferably selected from the group consisting of: (i) less than 0.5 mm; (ii)0.5-1.0 mm; (iii)1.0-1.5 mm; (iv)1.5-2.0 mm; (v)2.0-2.5 mm; (vi)2.5-3.0 mm; (vii)3.0-3.5 mm; (viii)3.5-4.0 mm; (ix)4.0-4.5 mm; (x)4.5-5.0 mm; (xi) Less than 5.0 mm; and (xii) > 5.0 mm.

According to a preferred embodiment, the tapered section of the aperture is preferably substantially linearly tapered. For example, the diameter of the circumference of the aperture preferably varies (e.g., decreases) substantially linearly with the depth of the aperture. If the aperture has a cross-sectional shape other than circular, then the cross-sectional dimension (e.g., the smallest span of the cross-sectional shape of the aperture) or the perimeter of the cross-sectional shape of the aperture varies (e.g., decreases) substantially linearly with the depth of the aperture.

According to a preferred embodiment, the reagent beads are preferably opaque and the signal is preferably taken only from the top of the beads. The bottom of the bead below the press-fit line is preferably not in contact with the fluid. In a preferred embodiment, in use, the reagent beads preferably form a substantially fluid tight seal with the tapered section of the bore, preferably to substantially prevent fluid flow from the sample well past the reagent beads. The sample plate with inserted reagent beads according to the preferred embodiment thus resembles a conventional sample well that is emptied in a rather close manner.

According to a preferred embodiment, the reagent beads do not protrude above the bottom of the sample well and are therefore preferably not prone to damage due to handling, pipetting or washing. However, a less preferred embodiment may be desired in which one or more reagent beads may protrude slightly above the bottom of the sample well.

According to a preferred embodiment, the depth of the aperture is preferably equal to or greater than the diameter of the reagent bead, so that the reagent bead does not protrude above the bottom of the sample well.

According to a preferred embodiment, the depth of the aperture is preferably selected from the group consisting of: (i) less than 0.5 mm; (ii)0.5-1.0 mm; (iii)1.0-1.5 mm; (iv)1.5-2.0 mm; (v)2.0-2.5 mm; (vi)2.5-3.0 mm; (vii)3.0-3.5 mm; (viii)3.5-4.0 mm; (ix)4.0-4.5 mm; (x)4.5-5.0 mm; (xi) Less than 5.0 mm; and (xii) > 5.0 mm.

According to a preferred embodiment, the depth of the aperture where the diameter of the aperture becomes smaller than the diameter of the reagent bead is preferably equal to or larger than the radius of the reagent bead, so that the reagent bead does not protrude above the bottom of the sample well. If the aperture has a cross-sectional shape other than circular, it is the depth of the aperture where the smallest span of the cross-sectional shape of the aperture becomes smaller than the diameter of the reagent bead that is preferably equal to or larger than the radius of the reagent bead.

According to a preferred embodiment, in use, the reagent beads are preferably not in contact with the base of the bore. However, less preferred embodiments are also contemplated in which the reagent beads do come into contact with the base of the bore

An advantageous aspect of the preferred embodiments is that since the reagent beads are preferably arranged to be inserted such that they are flush with the bottom of the wells, the sample plate according to the present invention can be used with known automated microplate processing systems without any hardware changes. Furthermore, the sample wells according to the preferred embodiments are columns of similar proportions to wells of conventional microplates, and thus the fluid or other operational characteristics of the sample wells are well known. The processing steps according to the preferred embodiment such as pipetting, mixing, washing and incubation preferably follow the same type of fluidic behaviour as experienced by conventional microplates.

The sample plate according to the preferred embodiment preferably has a fluid capacity of about 800 microliters but advantageously only about 300 microliters of fluid is needed in use to cover all reagent beads arranged in the sample plate base.

Another advantageous feature of the sample plate according to the preferred embodiment is that fluid can be dispensed directly into the centre of the sample well, and according to the preferred embodiment the sample plate may be arranged such that the pockets, recesses or apertures for fastening reagent beads are not arranged in the central region of the sample well. This arrangement is particularly advantageous because the reagent preferably covering the reagent beads is not inadvertently washed off the reagent beads by the force of the fluid jet from the shampoo or pipette tip.

The sample plate according to the preferred embodiment preferably enables multiple tests to be performed in a single sample well. This is achieved by inserting different reagent beads into separate apertures of the same sample well, thereby enabling multiplexing to be performed. According to a preferred embodiment, the reagent beads can be pressed into a tapered hole in the base of the well as needed, which results in a high degree of flexibility and the ability to use the entire sample well with high efficiency.

A sample plate according to one embodiment of the present invention may include one or more sample wells having a diameter of 12 mm. Each sample well may have a dimension of 58mm²And a total of 54 sample wells of this size can fit into a conventional microplate base (footprint). A variable number of beads can be inserted in each sample well. The tapered apertures can have different diameters to accommodate different sizes as desiredThe reagent bead of (1).

According to other embodiments, one or more sample wells may comprise a 6 x 3.0mm diameter cavity, a recessed or tapered aperture, a 10 x 2.0mm diameter cavity, a recessed or tapered aperture, or a 21 x 1.75mm cavity, recessed or tapered aperture. The central region of the sample well is preferably free of voids, depressions or tapered pore sizes. Void, recessed or tapered apertures. The voids, depressions, or tapered apertures can be arranged in one or more concentric circles or other patterns around the central region of the sample well.

According to one embodiment, a sample plate with a 9 x 6 array of sample wells may be provided. If 6 wells are provided per sample well, wells are recessed or tapered, then the sample plates can each hold 324 reagent beads. If 10 wells are provided per sample well, the wells are recessed or tapered, then the sample plates can each hold 540 reagent beads. If 21 wells are provided per sample well, the wells can each accommodate 1134 reagent beads.

According to a preferred embodiment, the sample plate according to the preferred embodiment is not subject to fluid mixing problems. The sample well preferably comprises a bead pressed or inserted into a hole, depression or tapered bore. Once the reagent beads are inserted, their top is preferably flush or level with the bottom of the sample well. According to a preferred embodiment, mixing uses a fluid above the surface of the bead to pull fluid from the void area around the bead.

A further advantageous aspect of the invention is that the sample plate according to the invention is relatively simple to manufacture compared to other known arrangements. The sample plate can be manufactured by molding using opening and closing tools, and thus manufacturability is high and reliable. The injection molding tool used to form the sample plate is simple in design and does not require the use of undercuts or thin parts for molding. Thus, production of sample plates of different formats can be easily achieved. A tool producing sample wells having 6 holes or apertures can be readily adapted to produce sample wells having different numbers (e.g., 21) of holes.

Another advantage of the preferred embodiments is that validation of different well designs and formats can be achieved simply because the test protocols are still substantially the same. The aspiration and incubation will not change and the washing process will at best require only minor changes to the aspiration path.

It is therefore clear that the sample plate according to the invention is particularly advantageous compared to other known sample plates, including the sample plate disclosed in US-5620853.

The tapered section preferably has an angle of from (i)2-4 °; (ii)4-6 degrees; (iii)6-8 degrees; (iv)8-10 degrees; (v) at least 1 °; and (vi) a taper selected from the group consisting of 1-15 °.

According to a less preferred arrangement, the void or depression provided in the base may comprise a cavity with a retaining element, membrane, lip or annular portion (optionally instead of an aperture with a tapered section). The reagent bead or microsphere may be inserted through or past the retaining element, membrane, lip or annular portion into the cavity in use and may be substantially retained or secured within the cavity by the retaining element, membrane, lip or annular portion.

The one or more pockets or recesses include a counter-sunk or enlarged portion to facilitate insertion of the reagent beads or microspheres into one or more of the pockets or recesses.

The one or more sample wells comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 pockets or recesses, each comprising a bore having a tapered section and each arranged and adapted to receive, in use, a reagent bead or microsphere.

The one or more holes, recesses or apertures provided in the base are arranged to: (i) circumferentially surrounding a central portion of the sample well; and/or (ii) a cavity or recess having a plurality of cavities or recesses circumferentially surrounding more than one central cavity or recess; and/or (iii) in a substantially close-packed (close-pack) arrangement; and/or (iv) in a substantially symmetrical or asymmetrical manner; and/or (v) in a substantially linear or curved manner; and/or (vi) in a substantially regular or irregular manner; and/or (vii) an array; and/or (viii) one or more concentric circles and no holes, depressions or apertures in the center of the base.

The sample plate is preferably made of polystyrene or otherwise.

The sample plate may comprise a format of strips or arrays. For example, according to a preferred embodiment, the sample plate may comprise 6 x 1 strips. According to another preferred embodiment, the sample plate may comprise 9 x 6 strips.

According to other embodiments, the sample plate may comprise sample wells arranged in an a x B format, wherein:

a is selected from the group consisting of (i) 1; (ii) 2; (iii) 3; (iv) 4; (v) 5; (vi) 6; (vii) 7; (viii) 8; (ix) 9; (x) 10; and (xi) > 10; and is

B is selected from the group consisting of (i) 1; (ii) 2; (iii) 3; (iv) 4; (v) 5; (vi) 6; (vii) 7; (viii) 8; (ix) 9; (x) 10; and (xi) > 10.

According to one embodiment, one or more of the sample wells may be interconnected to one or more other sample wells by one or more frangible regions or joints, so that the sample plate can be separated into a plurality of smaller sample plates by a user. This allows the sample plate to be broken or fragmented into a plurality of smaller sample plates. For example, a 6 x 1 strip sample plate may be broken off into individual 1 x 1 sample plates comprising a single sample well or into two sample plates each comprising 3 x 1 sample wells.

According to another arrangement, a sample plate is provided comprising a plurality of sample wells, wherein one or more of the sample wells comprises one or more central fluid receiving areas and a plurality of reagent bead receiving chambers arranged around the one or more central fluid receiving areas, wherein the one or more central fluid receiving areas are in fluid communication with at least some or all of the reagent bead receiving chambers.

The one or more sample wells may comprise a peripheral wall defining the plurality of reagent bead-receiving cavities together with a plurality of radial wall elements, wherein in use a reagent bead is received in the reagent bead-receiving cavity and is prevented from passing radially through into the central fluid-receiving region by the radial wall elements.

Fluid dispensed into the one or more central fluid receiving areas in use may flow into some or all of the reagent bead receiving chambers without overflowing from the peripheral wall and/or without overflowing from the plurality of radial wall elements.

One or more of the sample wells may be interconnected to one or more other sample wells by one or more frangible regions or joints to enable the sample plate to be separated into a plurality of smaller sample plates by a user.

The sample plate may comprise an immunoassay sample plate. Alternatively, the sample plate may comprise hybridization probes for detecting the presence of a complementary DNA or RNA sample.

The sample plate preferably comprises a base portion having a female, male or other interface for securing the sample plate to a corresponding male, female or other interface of the plate frame holder.

According to one aspect of the present invention there is provided a combination of a sample plate as described above and one or more reagent beads or microspheres inserted or positioned in one or more of the pockets, recesses or apertures of the one or more sample wells.

According to another arrangement, there is provided a combination of a sample plate as described above and one or more reagent beads or microspheres inserted or positioned in one or more of the reagent bead receiving chambers of the one or more sample wells.

At least some or substantially all of the reagent beads or microspheres preferably carry, include or are otherwise covered with a reagent, wherein the reagent is arranged and adapted to determine an analyte of interest in the sample liquid.

According to alternative embodiments, at least some or substantially all of the reagent beads or microspheres carry, include or are otherwise covered with nucleic acid probes arranged and adapted to hybridize to single stranded nucleic acids, DNA or RNA.

According to another aspect of the present invention there is provided a combination of a plate frame holder and a sample plate as described above.

The plate frame holder preferably comprises a male, female or other interface for securely fastening the sample plate to the plate frame holder.

According to one aspect of the present invention, there is provided an automated apparatus comprising:

one or more reagent bead or microsphere dispensers;

a sample plate as described above; and

a control system arranged and adapted to control dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more sample wells of a sample plate.

The one or more reagent bead or microsphere dispensers preferably comprise:

a syringe body comprising an annular chamber surrounding a longitudinal bore, wherein the annular chamber is arranged to direct or leak reagent beads or microspheres disposed in the annular chamber towards the chamber disposed in the bore, in use;

a plunger disposed in the longitudinal bore; and

a barrel or nozzle;

wherein the plunger is arranged to dispense the reagent beads or microspheres from the chamber into the barrel or nozzle in use.

According to one aspect of the present invention there is provided a device for determining one or more analytes of interest in a liquid, the device comprising:

one or more reagent bead or microsphere dispensers; and

a sample plate as described above.

According to one aspect of the present invention there is provided a reagent bead or microsphere dispenser for dispensing a reagent bead or microsphere into one or more pockets, recesses or bores of a sample well, the reagent bead or microsphere dispenser comprising:

a syringe body comprising an annular chamber surrounding a longitudinal bore, wherein the annular chamber is arranged to direct or leak reagent beads or microspheres disposed in the annular chamber towards the chamber disposed in the bore, in use;

a plunger disposed in the longitudinal bore; and

a barrel or nozzle;

wherein the plunger is arranged to dispense the reagent beads or microspheres from the chamber into the barrel or nozzle in use.

According to an aspect of the invention, there is provided a method comprising:

providing one or more reagent bead or microsphere dispensers;

providing a sample plate as described above; and

controlling dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more of the sample wells.

According to one aspect of the present invention, there is provided a method of analyzing a plurality of analytes in a sample using a sample plate, comprising:

providing a sample plate as described above; and

inserting one or more reagent beads or microspheres into one or more pockets, recesses, or apertures of a sample well; and

the sample is added to the sample well.

According to one aspect of the present invention, there is provided a method for detecting an antigen or an antibody in a sample using enzyme-linked immunosorbent assay (ELISA), comprising:

providing a sample plate as described above; and

inserting one or more reagent beads or microspheres into one or more pockets, recesses, or apertures of a sample well; and

the sample is added to the sample well.

According to one aspect of the present invention, there is provided a method for detecting a DNA or RNA sequence in a sample using a nucleic acid probe, comprising:

providing a sample plate as described above; and

inserting one or more reagent beads or microspheres into one or more pockets, recesses, or apertures of a sample well; and

the sample is added to the sample well.

According to one aspect of the present invention, there is provided a method for determining one or more analytes of interest in a sample, comprising:

inserting one or more reagent beads or microspheres into one or more pockets or recesses of one or more sample wells of a sample plate, wherein the one or more pockets or recesses comprise a bore having a tapered section.

According to one embodiment, the method preferably further comprises one or more of the following steps: (i) cultivating a sample plate; and/or (ii) washing the sample plate; and/or (iii) a aspirated sample plate; and/or (iv) adding an enzyme conjugate to the sample plate; and/or (v) adding a visualization agent to the sample plate; and/or (vi) visually analyzing the sample plate.

According to one aspect of the present invention, there is provided a kit for performing an enzyme-linked immunosorbent assay (ELISA) procedure, comprising:

one or more sample plates as described above; and

a plurality of reagent beads or microspheres covered with a reagent comprising an antibody, an antigen or another biomolecule.

According to another aspect of the present invention, there is provided a kit for a nucleic acid detection process, comprising:

one or more sample plates as described above; and

a plurality of reagent beads or microspheres covered with DNA or RNA sequences.

According to an aspect of the present invention, there is provided a method of manufacturing a sample plate, comprising:

providing a sample plate comprising one or more sample wells each having a base; and

forming one or more pockets or recesses in the one or more bases, wherein the one or more pockets or recesses comprise a bore having a tapered section and wherein the one or more pockets or recesses are arranged and adapted to receive, in use, a reagent bead or microsphere.

According to an aspect of the invention there is provided a computer program executable by a control system of an automated apparatus comprising one or more reagent bead or microsphere dispensers, the computer program being arranged to cause the control system to:

(i) controlling the dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more sample wells of a sample plate having one or more pockets or recesses comprising a bore having a tapered section.

According to an aspect of the invention there is provided a computer readable medium comprising computer executable instructions stored on the computer readable medium, the instructions being arranged to be executable by a control system of an automated apparatus, the automated apparatus comprising one or more reagent bead or microsphere dispensers, the computer program being arranged to cause the control system to:

(i) controlling the dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more sample wells of a sample plate having one or more pockets or recesses comprising a bore having a tapered section.

The computer readable medium is preferably selected from the group consisting of (i) a ROM; (ii) an EAROM; (iii) an EPROM; (iv) an EEPROM; (v) a fast memory; (vi) an optical disc; (vii) a RAM; and (viii) hard disk drives.

According to another arrangement, there is provided an apparatus comprising:

one or more reagent bead or microsphere dispensers;

a sample plate comprising a plurality of sample wells, wherein one or more of the sample wells comprises one or more central fluid receiving areas and a plurality of reagent bead or microsphere receiving cavities disposed about the one or more central fluid receiving areas, wherein the one or more central fluid receiving areas are in fluid communication with at least some or all of the reagent bead or microsphere receiving cavities; and

a control system arranged and adapted to control dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more of the plurality of reagent bead or microsphere receiving chambers.

Other embodiments may be desired in which the reagent bead or microsphere-receiving cavity may more broadly comprise only a reagent bead or microsphere-receiving region or location. Thus, the term "reagent bead or microsphere receiving chamber" may be replaced with the term "reagent bead or microsphere receiving area or location".

One or more of the sample wells preferably comprises a peripheral wall, surface or well, wherein fluid dispensed into the sample well is preferably confined within the sample well by the peripheral wall, surface or well.

The device preferably further comprises one or more wall elements, surfaces or grooves which preferably together with the above mentioned peripheral walls, surfaces or grooves define said plurality of reagent bead or microsphere receiving chambers.

Fluid dispensed into the one or more central fluid receiving areas in use preferably flows into some or all of the reagent bead or microsphere receiving cavities without overflowing the peripheral wall, surface or groove and/or overflowing the one or more wall elements, surfaces or grooves.

The one or more wall elements, surfaces or grooves together with a portion of the peripheral wall, surface or groove preferably define respective reagent bead or microsphere receiving cavities.

The one or more wall elements, surfaces or grooves preferably extend inwardly from the peripheral wall in a radial, linear or curved manner.

At least some or all of the wall elements, surfaces or grooves are preferably integral with or depending from the peripheral wall. According to an alternative arrangement, at least some or all of the wall elements, surfaces or grooves are radially spaced from the peripheral wall or are separated therefrom by a gap.

The peripheral wall, surface or groove preferably has a thickness of from (i) < 1 mm; (ii)1-2 mm; (iii)2-3 mm; (iv)3-4 mm; (v)4-5 mm; (vi)5-6 mm; (vii)6-7 mm; (viii)7-8 mm; (ix)8-9 mm; (x)9-10 mm; (xi)10-11 mm; (xii)11-12 mm; (xiii)12-13 mm; (xiv)13-14 mm; (xv)14-15 mm; (xvi)15-16 mm; (xvii)16-17 mm; (xviii)17-18 mm; (xix)18-19 mm; (xx)19-20 mm; and (xxi) > 20 mm.

The wall element, surface or groove preferably has a thickness of from (i) < 1 mm; (ii)1-2 mm; (iii)2-3 mm; (iv)3-4 mm; (v)4-5 mm; (vi)5-6 mm; (vii)6-7 mm; (viii)7-8 mm; (ix)8-9 mm; (x)9-10 mm; (xi)10-11 mm; (xii)11-12 mm; (xiii)12-13 mm; (xiv)13-14 mm; (xv)14-15 mm; (xvi)15-16 mm; (xvii)16-17 mm; (xviii)17-18 mm; (xix)18-19 mm; (xx)19-20 mm; and (xxi) > 20 mm.

At least some or substantially all of the plurality of reagent bead or microsphere receiving cavities are preferably arranged and adapted to receive, in use, a single reagent bead or microsphere or a plurality of reagent beads or microspheres.

According to one embodiment, at least some or substantially all of the sample wells comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or > 20 reagent bead or microsphere receiving cavities.

The sample well preferably comprises one or more reagent bead or microsphere receiving cavities that are circular, oval, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, decagonal, or polygonal.

According to one embodiment, one or more of the sample wells has a length of from (i) < 1 mm; (ii)1-2 mm; (iii)2-3 mm; (iv)3-4 mm; (v)4-5 mm; (vi)5-6 mm; (vii)6-7 mm; (viii)7-8 mm; (ix)8-9 mm; (x)9-10 mm; (xi)10-11 mm; (xii)11-12 mm; (xiii)12-13 mm; (xiv)13-14 mm; (xv)14-15 mm; (xvi)15-16 mm; (xvii)16-17 mm; (xviii)17-18 mm; (xix)18-19 mm; (xx)19-20 mm; and (xxi) > 20 mm.

The one or more fluid-receiving regions are preferably in fluid communication with one or more of the reagent bead or microsphere receiving cavities such that, in use, fluid received in the one or more fluid-receiving regions flows into the one or more reagent bead or microsphere receiving cavities.

The sample well preferably comprises one or more circular, elliptical, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, decagonal or polygonal fluid receiving areas.

The reagent beads or microspheres that are dispensed into one or more of the well, recess, bore or reagent bead or microsphere receiving chamber in use preferably have a particle size selected from the group consisting of (i) < 0.5 mm; (ii)0.5-1.0 mm; (iii)1.0-1.5 mm; (iv)1.5-2.0 mm; (v)2.0-2.5 mm; (vi)2.5-3.0 mm; (vii)3.0-3.5 mm; (viii)3.5-4.0 mm; (ix)4.0-4.5 mm; (x)4.5-5.0 mm; (xi) (xii) > 5.0mm and (xii) > 5.0 mm.

At least some or substantially all of the reagent beads or microspheres that are dispensed into one or more of the well, recess, bore or reagent bead or microsphere-receiving chamber in use may carry or comprise a reagent, wherein the reagent is arranged and adapted to: (i) analyzing the sample; and/or (ii) analyzing the sample by a nucleic acid amplification reaction; and/or (iii) analyzing the sample by Polymerase Chain Reaction (PCR); and/or (iv) analyzing the sample by an immunoassay process; and/or (v) analyzing the sample by using hybridization probe techniques.

At least some or substantially all of the reagent beads or microspheres that are dispensed into one or more of the well, recess, bore or reagent bead or microsphere-receiving chamber in use comprise polystyrene, plastic or polymer.

According to one embodiment, at least some or substantially all of the reagent beads or microspheres that are dispensed into one or more of the well, recess, bore, or reagent bead or microsphere-receiving cavity in use comprise or have an iron-based or magnetic coating.

At least some or substantially all of the reagent beads or microspheres that are dispensed into the well, recess, bore or one or more of the reagent bead or microsphere-receiving cavities in use preferably comprise an antistatic coating or have antistatic properties.

The apparatus preferably further comprises magnetic and/or electrostatic means arranged and adapted to: (i) drawing one or more reagent beads or microspheres into the plurality of pockets, recesses, apertures or reagent bead or microsphere receiving cavities as the reagent beads or microspheres are dispensed; and/or (ii) attract and/or retain one or more reagent beads or microspheres that have been dispensed into the plurality of pockets, recesses, apertures or reagent bead or microsphere receiving cavities such that the one or more reagent beads or microspheres are retained or retained in the pockets, recesses, apertures or reagent bead or microsphere receiving cavities for at least a period of time.

According to one embodiment, the apparatus further comprises a mechanical device and/or an electrical device arranged and adapted to: (i) directing one or more reagent beads or microspheres as they are dispensed such that the one or more reagent beads or microspheres are received in the plurality of reagent bead or microsphere receiving chambers; and/or (ii) retaining one or more reagent beads or microspheres that have been dispensed into the plurality of reagent bead or microsphere receiving cavities such that the one or more reagent beads or microspheres are retained or retained in the reagent bead or microsphere receiving cavities for at least a period of time.

The apparatus preferably further comprises magnetic and/or electrostatic means arranged and adapted to: vibrating and/or agitating one or more reagent beads or microspheres that have been received in the plurality of reagent bead or microsphere receiving chambers.

The apparatus preferably further comprises mechanical and/or electrical means arranged and adapted to: vibrating and/or agitating one or more reagent beads or microspheres that have been received in the plurality of reagent bead or microsphere receiving chambers.

According to one embodiment, one or more of the reagent bead or microsphere dispensers preferably comprises a tube containing a plurality of reagent beads or microspheres in use.

One or more of the reagent bead or microsphere dispensers preferably includes a screw, auger or reagent bead or microsphere delivery device for delivering or delivering one or more reagent beads or microspheres within the reagent bead or microsphere dispenser to a dispensing region, dispensing end or dispensing tip of the reagent bead or microsphere dispenser.

The apparatus preferably further comprises one or more sensors for sensing whether one or more reagent beads have been dispensed from one or more of the reagent bead or microsphere dispensers.

The apparatus preferably further comprises a translation stage for moving the sample plate relative to the one or more reagent bead or microsphere dispensers.

The control system is preferably arranged and adapted to control the translation stage such that one or more reagent beads or microspheres from the reagent bead or microsphere dispenser are sequentially dispensed into different reagent bead or microsphere receiving chambers by moving the sample plate relative to the reagent bead or microsphere dispenser.

The apparatus preferably further comprises a rotatable carousel, wherein the one or more reagent bead or microsphere dispensers are attached or attachable to the carousel.

The control system is preferably arranged and adapted to rotate the carousel after all desired first reagent beads or microspheres have been dispensed from the first reagent bead or microsphere dispenser into the plurality of different reagent bead or microsphere receiving cavities, wells, recesses or apertures of the sample plate such that a second different reagent bead or microsphere dispenser is then brought into a position where the second reagent bead or microsphere dispenser is able to dispense a second reagent bead or microsphere into the plurality of different reagent bead or microsphere receiving cavities, wells, recesses or apertures of the sample plate. This process is then preferably repeated for other (e.g., third, fourth, fifth, sixth, seventh, eighth, etc.) reagent bead or microsphere dispensers.

According to one embodiment, the apparatus further comprises a fluid dispensing device to dispense fluid into one or more fluid receiving regions of one or more sample wells.

The fluid dispensing device is preferably arranged and adapted to dispense x millilitres of fluid at a time into said one or more fluid receiving areas of one or more sample wells, wherein x is preferably selected from the group consisting of (i) < 10; (ii)10-20 parts of; (iii)20-30 parts of; (iv)30-40 parts of; (v) 40-50; (vi)50-60 parts of; (vii)60-70 parts of; (viii)70-80 parts; (ix) 80-90; (x)90-100 parts of; (xi) 100-; (xii) 110-120; (xiii) 120-130; (xiv) 130-140; (xv) 140-150; (xvi) 150-; (xvii) 160-170; (xviii)170- > 180; (xix) 180-190; (xx) 190-200; and (xxi) > 200.

According to one embodiment, the apparatus further comprises an image analysis device or camera to determine whether a reagent bead or microsphere has been dispensed or is otherwise present in the reagent bead or microsphere receiving cavity, well, recess or aperture.

The sample plate preferably has a first color and the reagent bead or microsphere preferably has a second, different color that contrasts with the first color to facilitate visual detection of the presence or absence of the reagent bead or microsphere in the reagent bead or microsphere receiving cavity, well, depression or aperture.

According to one embodiment, the sample plate may further comprise luminescent or fluorescent labels.

The apparatus may also include a luminescent or fluorescent detection device to determine whether a reagent bead or microsphere has been dispensed or is otherwise present in the reagent bead or microsphere-receiving cavity, well, depression or aperture by determining whether the reagent bead or microsphere obstructs or partially obstructs the luminescent or fluorescent label.

The device preferably further comprises a magnetic and/or electrical and/or capacitive and/or mechanical sensor to sense whether reagent beads or microspheres have been dispensed or are otherwise present in the reagent bead or microsphere receiving cavity, well, recess or aperture of the sample plate.

The control system preferably determines the number of reagent beads or microspheres present in the sample well and/or the number of missing reagent beads or microspheres and/or the number of reagent beads or microspheres dispensed and/or the number of reagent beads or microspheres desired to be dispensed.

According to one embodiment, the control system measures and/or adjusts the volume of fluid dispensed or desired to be dispensed into the sample well in dependence on the number of reagent beads or microspheres determined to be present and/or absent and/or dispensed and/or desired to be dispensed in the sample well.

The control system is preferably arranged and adapted to ensure that at least some or substantially all of the reagent beads or microspheres in the sample well are at least partially or fully submerged by the fluid as the fluid is dispensed into the sample well.

The control system is preferably arranged and adapted to ensure that the height of fluid dispensed into the sample well remains substantially constant irrespective of the number of reagent beads or microspheres present, absent, dispensed or desired to be dispensed into the sample well.

According to another arrangement there is provided a combination of a device as described above together with a plurality of reagent beads or microspheres positioned in the one or more reagent bead or microsphere dispensers and/or the one or more reagent bead or microsphere receiving cavities, recesses or apertures.

According to another arrangement, there is provided a method comprising:

providing one or more reagent bead or microsphere dispensers;

providing a sample plate comprising a plurality of sample wells, wherein one or more of the sample wells comprises one or more central fluid receiving areas and a plurality of reagent bead or microsphere receiving cavities disposed about the one or more central fluid receiving areas, wherein the one or more central fluid receiving areas are in fluid communication with at least some or all of the reagent bead or microsphere receiving cavities; and

controlling the dispensing of reagent beads or microspheres from the one or more reagent bead or microsphere dispensers into one or more of the plurality of reagent bead or microsphere receiving chambers.

According to another arrangement, there is provided a sample plate comprising a plurality of sample wells, wherein one or more of the sample wells comprises one or more central fluid receiving areas and a plurality of reagent bead or microsphere receiving chambers arranged around the one or more central fluid receiving areas, wherein the one or more central fluid receiving areas are in fluid communication with at least some or all of the reagent bead or microsphere receiving chambers.

One or more of the sample wells preferably comprises a peripheral wall which together with a plurality of radial wall elements defines the plurality of reagent bead or microsphere receiving chambers, wherein in use a reagent bead or microsphere is received in the reagent bead or microsphere receiving chamber and is prevented from radially entering the central fluid receiving area by the radial wall elements.

The one or more radial wall elements are preferably integral with the peripheral wall or spaced from the peripheral wall by a gap.

The one or more radial wall elements preferably comprise one or more protrusions which preferably help to confine the reagent bead or microsphere within the reagent bead or microsphere receiving cavity and/or preferably help to prevent the reagent bead or microsphere from entering the central fluid receiving area radially.

According to one embodiment, the radial wall element has a diameter from (i) < 1 mm; (ii)1-2 mm; (iii)2-3 mm; (iv)3-4 mm; (v)4-5 mm; (vi)5-6 mm; (vii)6-7 mm; (viii)7-8 mm; (ix)8-9 mm; (x)9-10 mm; (xi)10-11 mm; (xii)11-12 mm; (xiii)12-13 mm; (xiv)13-14 mm; (xv)14-15 mm; (xvi)15-16 mm; (xvii)16-17 mm; (xviii)17-18 mm; (xix)18-19 mm; (xx)19-20 mm; and (xxi) > 20 mm.

The one or more sample wells preferably comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or > 20 reagent bead or microsphere receiving cavities.

The sample well preferably comprises one or more reagent bead or microsphere receiving cavities that are circular, oval, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, decagonal, or polygonal.

The one or more sample wells have a length of from (i) < 1 mm; (ii)1-2 mm; (iii)2-3 mm; (iv)3-4 mm; (v)4-5 mm; (vi)5-6 mm; (vii)6-7 mm; (viii)7-8 mm; (ix)8-9 mm; (x)9-10 mm; (xi)10-11 mm; (xii)11-12 mm; (xiii)12-13 mm; (xiv)13-14 mm; (xv)14-15 mm; (xvi)15-16 mm; (xvii)16-17 mm; (xviii)17-18 mm; (xix)18-19 mm; (xx)19-20 mm; and (xxi) > 20 mm.

The sample well preferably comprises one or more circular, elliptical, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, decagonal or polygonal fluid receiving areas.

Drawings

Various embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which:

FIG. 1 shows a first principal embodiment of the invention in which a plurality of reagent bead or microsphere dispensers are attached to a rotatable carousel and a sample plate is mounted on a translation stage under an arm extending from the rotatable carousel and engaging the reagent bead or microsphere dispensers;

FIG. 2 shows a reagent bead or microsphere dispenser according to a first main embodiment of the present invention;

FIG. 3 shows in more detail a plurality of reagent bead or microsphere dispensers mounted to a carousel and an arm engaged with the reagent bead or microsphere dispensers according to a first main embodiment of the present invention;

FIG. 4A shows a first configuration of sample wells of a sample plate according to a first main embodiment of the invention; FIG. 4B shows a second, different configuration of a sample well of a sample plate according to another embodiment of the invention and FIG. 4C shows how different kinds or types of reagent beads or microspheres may be dispensed into different reagent bead or microsphere receiving cavities or sections of a sample well of a sample plate according to one embodiment of the invention;

FIG. 5 shows a sample well according to a first main embodiment of the invention;

figure 6 shows a sample well of a sample plate according to a second main embodiment of the invention;

FIG. 7A shows a plan view of a sample well of a sample plate according to the second main embodiment, FIG. 7B shows the bottom of the sample well according to the second main embodiment in more detail and FIG. 7C shows a reagent bead or microsphere dispensed in a cavity of the sample well according to the second main embodiment;

FIG. 8A shows a reagent bead or microsphere dispenser according to a second main embodiment of the present invention and FIG. 8B shows a cross-sectional view of the reagent bead or microsphere dispenser;

FIG. 9 shows an exploded view of a reagent bead or microsphere dispenser according to a second main embodiment;

FIG. 10 shows a microarray spotter according to a second main embodiment of the present invention comprising a reagent bead or microsphere injector pick-up device mounted on an x-y-z translation stage and engaged with a reagent bead or microsphere dispenser above a sample plate;

FIG. 11 shows in more detail a cross-sectional view of a reagent bead or microsphere syringe pick-up device attached to a reagent bead or microsphere dispenser according to a second main embodiment of the present invention;

FIG. 12A shows a reagent bead or microsphere dispenser transported by a reagent bead or microsphere injector picking apparatus and FIG. 12B shows a reagent bead or microsphere in the process of being dispensed from the reagent bead or microsphere dispenser by a plunger mechanism actuated by the reagent bead or microsphere injector picking apparatus;

FIG. 13A shows a reagent bead or microsphere injector during discharge from a reagent bead or microsphere injector picking apparatus and FIG. 13B shows a reagent bead or microsphere injector that has been discharged from a reagent bead or microsphere injector picking apparatus;

fig. 14A shows 9 sample plates loaded into a plate frame, wherein each sample plate comprises one 6 sample well and fig. 14B shows a plate frame into which one or more sample plates may be loaded;

figure 15A shows one 6 sample wells in more detail and figure 15B shows one 6 sample wells being loaded into the plate frame;

fig. 16A shows a single sample well being loaded into a plate frame, fig. 16B shows two sample wells connected by a disconnect structure in more detail, fig. 16C shows a sample well having an end structure and fig. 16D shows a sample well having an ID and an orientation tag;

FIG. 17A shows the underside of one sample well, FIG. 17B shows a female alignment and retention member that helps align one sample well with the plate frame and FIG. 17C shows a corresponding male alignment and retention member provided in the base of the plate frame; and

figure 18 shows a cross-sectional view of one sample well and shows that the sample well has multiple tapered apertures with the angle of the taper being 6.0 deg. according to a preferred embodiment.

Detailed Description

A first main embodiment of the present invention will now be described in more detail with reference to fig. 1. According to a first main embodiment, a rotatable carousel 1 is preferably provided, comprising a plurality of abutments or segments arranged around the outer circumference or perimeter of the carousel 1. According to the particular embodiment shown in fig. 1, 24 abutments are provided, although other embodiments are contemplated in which a different number of abutments is provided. For example, according to other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, 26, 27, 28, 29, 30, or > 30 abutments may be provided.

A plurality of reagent bead or microsphere dispensers 2 are preferably attached or otherwise secured in use to carousel 1 at some or all of the interfaces. Each abutment preferably comprises an upper jaw 3 and a lower retaining pin 4. An upper cartridge 3 and a lower retaining pin 4 are preferably used to secure the reagent bead or microsphere dispenser 2 to the dock. Other embodiments may be desired in which the retaining pin 4 may be disposed in an upper position and the collet 3 may be disposed in a lower position.

A single reagent bead or microsphere dispenser 2 is shown in more detail in figure 2. The reagent bead or microsphere dispenser 2 preferably comprises a tubular body 5 having a lower funnel-shaped dispensing portion 6 and an upper cap portion 7. Each reagent bead or microsphere dispenser 2 is preferably filled with a plurality of reagent beads or microspheres in use. According to one embodiment, 2000 reagent beads or microspheres, each 1.75mm in diameter, may be loaded into a single reagent bead or microsphere dispenser 2. Other embodiments may be desired in which the capacity of the reagent bead or microsphere dispenser 2 is greater or less.

According to another embodiment, the reagent bead or microsphere dispenser 2 may be arranged to handle reagent beads or microspheres having a diameter other than 1.75 mm. A less preferred embodiment may be desired in which the reagent beads or microspheres in a first reagent bead or microsphere dispenser 2 may have a first diameter and in which the reagent beads or microspheres in a second, different reagent bead or microsphere dispenser 2 may have a second, different diameter. It is also contemplated that other less preferred embodiments may have multiple diameters or a mixture of different diameters of the reagent beads or microspheres loaded into a particular reagent bead or microsphere dispenser 2.

At least some of the reagent bead or microsphere dispensers 2 preferably comprise hooks 8, which preferably hang from the funnel-shaped dispensing portion 6 and are preferably arranged to connect or latch with the retaining pins 4 of the docking portion on the carousel 1. The upper part of the tubular body 5 is preferably arranged to be fastened to the butt joint by the collet 3 of the butt joint. The upper collets 3 of at least some of the abutments may take a different form to that shown in figure 1. Other embodiments may be desired in which different ways may be used to secure the reagent bead or microsphere dispenser 2 to the docking portion of the carousel 1.

Each reagent bead or microsphere dispenser 2 preferably includes a central auger, screw or screw mechanism 9 which, when rotated, preferably moves the reagent beads or microspheres from within the tubular body 5 towards the dispensing section 6. The base of the tubular body 5, which in use retains the reagent beads or microspheres, preferably comprises an annular disc or base section having a central aperture. An auger, screw or screw mechanism 9 preferably passes through a central aperture in the base of the tubular body 5. The dispensing portion 6 preferably comprises a tubular bore through which an auger, screw or screw mechanism 9 passes. The diameter of the tubular bore in the dispensing section 6 and the pitch of the auger, screw or screw mechanism 9 are preferably arranged so that the reagent beads or microspheres in the dispensing section 6 are urged towards the nozzle of the dispensing section 6 and can be dispensed from the dispensing section 6 one at a time.

The shaft or upper end of the auger, screw or screw mechanism 9 is preferably connected to a first gear or other first drive mechanism 10. Referring to fig. 1, a first gear or other drive mechanism 10 at the upper end of the auger, screw or screw mechanism 9 is preferably arranged to be driven by a corresponding second drive gear 11 or second drive mechanism, preferably arranged on an arm 12 of the carousel 1. The teeth on the first gear 10 of the reagent bead or microsphere dispenser 2 preferably mesh with and interlock with corresponding teeth on the second drive gear 11 on the arm 12 of the carousel 1, such that rotation of the second drive gear 11 on the arm 12 of the carousel 1 causes rotation of the first gear 10 and hence rotation of the auger, screw or screw mechanism 9 connected to the first gear 10.

According to one embodiment of the invention, each reagent bead or microsphere dispenser 2 is preferably filled with a plurality of reagent beads or microspheres. The reagent beads or microspheres preferably comprise a polystyrene, plastic or polymer core, which is preferably covered with an iron-based or magnetic coating or which has iron-based or magnetic properties. The reagent beads or microspheres may be coated with a reagent (e.g., an antibody or antigen) that is preferably used to analyze the sample. According to one embodiment, the reagents may be used to analyze a sample by Polymerase Chain Reaction (PCR) or as part of an immunoassay process. Alternatively, according to an equally preferred embodiment, the reagent may comprise a DNA or RNA sequence which is used as a hybridization probe to detect the presence of complementary DNA or RNA sequences in the sample. The reagent beads or microspheres may also be covered with an antistatic coating or may have antistatic properties.

According to one embodiment, one or more sensors may be arranged on the carousel 1, preferably below or near the dispensing portion 6 of the reagent bead or microsphere dispenser 2. The one or more sensors preferably monitor whether one or more reagent beads or microspheres have been dispensed from the dispensing section 6 into the reagent bead or microsphere receiving chamber of the sample plate 13. The pitch of the auger, screw or screw mechanism 9 and the rotational speed of the auger, screw or screw mechanism 9 are preferably such that individual reagent beads or microspheres can be dispensed from the dispensing portion 6 of the reagent bead or microsphere dispenser 2 in less than 0.5 seconds.

As shown in fig. 1, the sample plate 13 is preferably mounted on a translation stage below the arm 12 of the carousel 1. The sample plate 13 preferably comprises a plurality of sample wells. Each sample well preferably comprises a central fluid receiving region and a plurality of reagent bead or microsphere receiving chambers disposed about the central fluid receiving region. The reagent beads or microspheres from the reagent bead or microsphere dispenser 2 are preferably dispensed into the desired reagent bead or microsphere receiving chamber in the sample plate 13. The sample plate 13 is preferably translated by a translation stage such that the desired reagent bead or microsphere receiving chamber is positioned in close proximity to the nozzle of the dispensing portion 6 of the reagent bead or microsphere dispenser 2. The reagent beads or microspheres are then dispensed into a reagent bead or microsphere receiving chamber and the sample plate 13 is moved by the translation stage such that the different reagent bead or microsphere receiving chambers are arranged in close proximity to the nozzle of the reagent bead or microsphere dispenser 2. The process of dispensing reagent beads or microspheres and translating the sample plate 13 is then preferably repeated. Once all the desired reagent beads or microspheres from a particular reagent bead or microsphere dispenser 2 have been dispensed into the appropriate reagent bead or microsphere receiving chamber of the sample plate 13, the carousel 1 is then preferably rotated to bring a second desired reagent bead or microsphere dispenser 2 into engagement with a second drive gear 11 arranged on the arm 12 of the carousel 1. The reagent beads or microspheres from the second reagent bead or microsphere dispenser 2 are then preferably dispensed into the desired reagent bead or microsphere receiving chamber of the sample plate 13. This process is preferably repeated so that reagent beads or microspheres from the further reagent bead or microsphere dispenser 2 attached to the carousel 1 are preferably dispensed into the further reagent bead or microsphere receiving chamber in the sample plate 13. Less preferred embodiments may also be desired, wherein some of the reagent bead or microsphere dispensers 2 attached to the carousel 1 may be replaced or supplemented during the process of dispensing reagent beads or microspheres into the sample plate 13.

A particularly advantageous feature is that reagent beads or microspheres may be dispensed from the reagent bead or microsphere dispenser 2 into the reagent bead or microsphere receiving cavity of the sample plate 13 in any desired manner. For example, in one sample well, reagent beads or microspheres of the same species or type may be dispensed into all of the reagent bead or microsphere receiving chambers. In another sample well, pairs of reagent beads or microspheres of the same species or type may be dispensed into adjacent reagent bead or microsphere receiving chambers. According to a preferred embodiment, a single reagent bead or microsphere is dispensed into each reagent bead or microsphere receiving chamber and a different type of reagent bead or microsphere is dispensed into each reagent bead or microsphere receiving chamber of a particular sample well. However, according to a less preferred embodiment, some of the reagent bead or microsphere receiving cavities may be empty. It may also be desirable, according to other less preferred embodiments, that some reagent bead or microsphere receiving chambers may receive more than one reagent bead or microsphere, particularly if the reagent bead or microsphere has a relatively small diameter relative to other reagent beads or microspheres that may be dispensed into other reagent bead or microsphere receiving chambers.

Figure 3 shows in more detail a plurality of reagent bead or microsphere dispensers 2 fastened by a collet 3 and a retaining pin 4 to a docking on a carousel 1. The retaining pin 4 preferably engages with a hook 8 provided on the dispensing portion 6 of the reagent bead or microsphere dispenser 2. The retaining pin 4, hook 8 and collet 3 preferably prevent the body of the reagent bead or microsphere dispenser 2 from rotating during use. The auger, screw or screw mechanism 9 within each reagent bead or microsphere dispenser 2 is preferably rotated or driven by bringing the teeth of a first gear 10 attached to the main shaft or shaft of the auger, screw or screw mechanism 9 into engagement or interlocking with a second drive gear or second drive mechanism 11, preferably suspended from an arm 12 of the carousel 1. The second drive gear or second drive mechanism 11 is preferably driven or rotated by an electric motor.

Fig. 4A shows individual sample wells 14 of the sample plate 11. According to the particular embodiment shown in fig. 4A, the sample well 14 may comprise 8 reagent bead or microsphere receiving chambers 15 arranged around a central fluid receiving area 16. Other embodiments may be desired in which a different number of reagent bead or microsphere receiving chambers or regions 15 are provided. Each reagent bead or microsphere receiving cavity 15 is preferably defined by at least two radial wall elements 17 together with the outer or inner wall of the sample well 14. The radial wall element 17 preferably hangs from the wall of the sample well 14 and preferably extends towards the centre of the sample well 14. However, the wall element 17 preferably does not extend all the way to the centre of the sample well 14 so that a central circular fluid receiving area 16 is preferably provided. At least some, or preferably all, of the radial wall elements 17 which preferably terminate adjacent the central fluid receiving region 16 may comprise an enlarged portion which is preferably designed to assist in retaining the reagent beads or microspheres within their respective reagent bead or microsphere receiving chamber 15 and to prevent the reagent beads or microspheres from entering the fluid receiving region 16. Other less preferred embodiments may be desired in which the height of at least some or substantially all of the radial wall elements 17 is reduced only at the location of the central fluid receiving area 16.

In the embodiment shown in fig. 4A, the radial wall element 17 is suspended from the outer or inner wall of the sample well 14. However, other embodiments are contemplated, such as the embodiment shown in fig. 4B, wherein the radial wall elements 17 are not suspended from the walls of the sample well 14. Alternatively, the radial wall element 17 is spaced from the outer or inner wall of the sample well 14. At least some, and preferably all, of the radial wall elements 17 terminating short of the outer or inner wall of the sample well 14 may comprise enlarged portions which are preferably designed to assist in retaining the reagent beads or microspheres within their respective reagent bead or microsphere receiving cavities 15. Other less preferred embodiments may be desired in which the height of at least some or substantially all of the radial wall elements 17 only decreases towards the outer or inner wall of the sample well 14.

Figure 4C shows an embodiment in which 8 different types of reagent beads or microspheres are shown as separate reagent bead or microsphere receiving chambers 15 dispensed into the sample well. In the particular embodiment shown in FIG. 4C, a first reagent bead or microsphere 18A is coated with a first reagent, a second reagent bead or microsphere 18B is coated with a second different reagent, a third reagent bead or microsphere 18C is coated with a third different reagent, a fourth reagent bead or microsphere 18D is coated with a fourth different reagent, a fifth reagent bead or microsphere 18E is coated with a fifth different reagent, a sixth reagent bead or microsphere 18F is coated with a sixth different reagent, a seventh reagent bead or microsphere 18G is coated with a seventh different reagent, and an eighth reagent bead or microsphere 18H is coated with an eighth different reagent. Thus, according to this embodiment, eight separately selected and distinct immunoassay processes can be performed substantially simultaneously on a single fluid sample, to enable multiplexed testing to be performed.

Fig. 5 shows a further embodiment of the invention, in which a sample plate comprising one strip of 6 sample wells 14 is provided. Each sample well 14 preferably comprises 8 reagent bead or microsphere receiving chambers 15.

Although the sample well 14 according to the first main embodiment preferably comprises a plurality of radial or straight walls 17, it is contemplated that in other embodiments the walls separating adjacent reagent beads or microsphere-receiving cavities 15 may be curved. According to yet another embodiment, the reagent bead or microsphere-receiving cavity 15 may have a honeycomb structure formed by a plurality of polygonal (e.g., hexagonal) cavities and/or may include a plurality of circular reagent bead or microsphere-receiving cavities 15.

According to the first main embodiment, the fluid to be tested is preferably dispensed into the central fluid receiving area 16 of the sample well 14. The fluid may, for example, comprise a blood, serum, saliva or urine sample taken from a patient. Fluid dispensed into the central fluid receiving region 16 of the sample well 14 preferably flows into each adjacent reagent bead or microsphere receiving chamber 15 by flowing between the gap between the two radial wall elements which help define the reagent bead or microsphere receiving chamber 15. According to a preferred embodiment, the dispensed fluid preferably does not overflow the top of the radial wall element 17.

At least some of the reagent beads or microspheres preferably dispensed into the reagent bead or microsphere-receiving cavity 15 of the sample well 14 may have an iron-based or magnetic layer or coating and/or have iron-based or magnetic properties. Magnetic or electrostatic devices may be used to attract reagent beads or microspheres as they are dispensed from the reagent bead or microsphere dispenser 2 in order to direct the reagent beads or microspheres 2 being dispensed into the appropriate reagent bead or microsphere receiving chamber 15 of the sample well 14. Once the reagent beads or microspheres have been dispensed into the reagent bead or microsphere receiving cavity 15, a magnetic or electrostatic device may then be used to attract, retain or otherwise maintain the reagent beads or microspheres within their reagent bead or microsphere receiving cavity 15 for a period of time.

Other embodiments may be desired in which mechanical or electrical devices may be used to leak or direct reagent beads or microspheres into the appropriate reagent bead or microsphere receiving chamber 15 and/or to retain or otherwise maintain reagent beads or microspheres that have been dispensed into the reagent bead or microsphere receiving chamber 15 within its chamber 15 for a period of time.

According to yet another embodiment, a magnetic, electrostatic, mechanical or electrical device may be used to vibrate or agitate the reagent beads or microspheres that have been dispensed into the suitable reagent bead or microsphere receiving chamber 15. According to one embodiment, the reagent beads or microspheres located in the reagent bead or microsphere-receiving chamber 15 may be vibrated or agitated once the fluid sample has been dispensed into the central fluid-receiving region 16 and once the fluid sample has been dispensed into each of the various reagent bead or microsphere-receiving chambers 15. This treatment helps to ensure that the various reagent beads or microspheres are completely wetted or otherwise covered by the dispensed fluid sample. According to one embodiment, 10-200ml of a fluid sample may be dispensed into each central fluid receiving area 16 of the sample wells 14 constituting the sample plate 13.

In addition to or as an alternative to sensors arranged on carousel 1 or otherwise closely adjacent to the dispensing portion 6 of reagent bead or microsphere dispenser 2, a visual detection system may be used to determine whether one or more reagent beads or microspheres have been dispensed or are otherwise correctly positioned in the appropriate reagent bead or microsphere receiving chamber 15 of sample plate 13. According to one embodiment, the reagent beads or microspheres may be colored and may contrast with the substantially clear color of the sample plate 13 according to one embodiment. The sample plate 13 may include one or more luminescent or fluorescent labels and a luminescent or fluorescent detection device may be used to determine whether a reagent bead or microsphere has been properly dispensed into the appropriate reagent bead or microsphere receiving cavity 15 of the sample well 14. This may be determined, for example, by determining whether the reagent beads or microspheres obscure or obstruct the view or otherwise detect a luminescent or fluorescent label on the sample plate 13. Other less preferred embodiments may be desired in which magnetic, electrical, capacitive or mechanical sensors may be used to determine the presence or absence of reagent beads or microspheres in the reagent bead or microsphere receiving cavity 15 of the sample plate 14.

According to one embodiment, the control system may be used to determine the number and/or location and/or type of reagent beads or microspheres that have been dispensed into the reagent bead or microsphere receiving chamber 15. The control system may also determine into which reagent bead or microsphere receiving chambers 15 further reagent beads or microspheres should be dispensed. Once the sample fluid has been dispensed into the central fluid receiving area of the sample well 14, the control system may check that an appropriate amount of sample fluid has been dispensed and that all of the reagent beads or microspheres are at least partially or completely submerged by the sample fluid.

The amount of sample fluid to be dispensed into the central fluid receiving region of the sample well 14 may depend on the number of reagent beads or microsphere receiving cavities 15 formed within the sample well 14, the diameter of the reagent beads or microspheres dispensed into the reagent bead or microsphere receiving cavities 15, and the number of reagent beads or microspheres dispensed into any given sample well 14. The control system may be used to vary the amount of sample fluid dispensed into the sample well 14 such that the depth of immersion of the reagent beads or microspheres in the sample fluid is substantially constant regardless of the number of reagent beads or microspheres in the sample well 14, the number of reagent bead or microsphere receiving cavities 15, and the diameter of the reagent beads or microspheres dispensed.

Different forms of sample plate 13 may be provided. For example, as shown in fig. 1 and 3, the sample plate 13 may include a two-dimensional array of sample wells 14. For example, the sample plate 13 may comprise a 4 × 4, 4 × 6, 4 × 8, 4 × 10, 4 × 12, 6 × 6, 6 × 8, 6 × 10, 6 × 12, 8 × 8, 8 × 10, 8 × 12, 10 × 10, 10 × 12, or 12 × 12 array of sample wells 14. According to other embodiments, the sample plate 13 may comprise one-dimensional strip-like sample wells 14. For example, the sample plate 13 may comprise 4 × 1, 6 × 1, 8 × 1, 10 × 1, or 12 × 1 sample wells 14. Other embodiments may be desired in which the sample wells 14 are arranged in other than an array or strip.

A second main embodiment of the present invention will now be described with reference to fig. 6. According to a second main embodiment, a sample plate is provided which preferably comprises a plurality of sample wells 19 (although according to another less preferred embodiment, a sample plate comprising only a single sample well 19 may be provided). According to a preferred embodiment, the sample plate may comprise a 9 x 6 array of sample wells 19. A single sample well 19 is shown in figure 6. Embodiments are also contemplated in which the sample plate may comprise one strip of sample wells 19, e.g. the sample plate may comprise a 1 x 9 or 1 x 6 array or strip of sample wells 19, for example.

Each sample well 19 preferably includes a plurality of pockets, recesses or apertures 21, which are preferably provided in the base of the sample well 19. In the particular embodiment shown in FIG. 6, the sample well 19 includes 10 pockets, recesses, or apertures 21 formed or otherwise provided in the base of the sample well 19. Other embodiments may be desired in which a different number of pockets, recesses, or apertures 21 may be provided in the base of the sample well 19. For example, according to alternative embodiments, at least some or all of the sample wells 19 provided in the sample plate may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or > 20 holes, recesses, or apertures 21.

The voids, depressions or apertures 21 are preferably disposed around the edge or periphery of the sample well 19 and the central or central region of the base of the sample well 19 is preferably substantially flat and free of voids, depressions or apertures 21. According to the first main embodiment described above with reference to fig. 1-5, the sample plate comprises a plurality of radial wall elements to retain the reagent beads or microspheres in their respective reagent bead or microsphere receiving chambers. However, according to the second main embodiment, the reagent beads or microspheres are preferably fastened within the cavities, recesses or apertures 21 of the sample plate 19 and therefore no radial wall elements are required and thus preferably no radial wall elements are provided. However, a less preferred embodiment may be desired in which the features of the first and second main embodiments are combined such that the sample plate is provided as comprising a plurality of reagent bead or microsphere receiving chambers defined in part by a plurality of radial wall elements. At least some of the reagent bead or microsphere-receiving cavities may further comprise a void, depression, or aperture provided in the base of the reagent bead or microsphere-receiving cavity. According to this less preferred embodiment, the reagent bead or microsphere may be dispensed into a reagent bead or microsphere receiving chamber or the reagent bead or microsphere may be securely fastened in a pocket, recess or aperture provided in the base of the reagent bead or microsphere receiving chamber.

Other embodiments in which a hybrid between a conventional microplate and a sample plate according to the first and/or second main embodiment is provided may also be desired. For example, according to one embodiment, a sample plate may be provided that includes one or more conventional sample wells and one or more sample wells having pockets, recesses, or apertures for receiving reagent beads or microspheres.

Referring to the second main embodiment as shown in fig. 6, at least some or all of the pockets, recesses or apertures 21 provided in the base of the sample well 19 preferably comprise apertures which are preferably tapered along at least a portion or substantially all of their length. The holes, recesses or apertures 21 may for example be arranged with a 6 deg. taper. According to one embodiment, the top of the tapered bore (or the reagent bead or microsphere-receiving portion) may have a diameter of 1.82 mm. The base of the sample well 19 surrounding the aperture may be arranged with a countersunk portion to facilitate insertion of the reagent beads or microspheres 20A, 20B into the pocket, recess or aperture 21. According to one embodiment, the outside diameter of the countersink may be 2.25 mm.

Fig. 7A shows a plan view of a sample well 19 and a part of two adjacent sample wells 19 provided in a sample plate according to a second main embodiment of the invention. The sample wells shown in fig. 7A form part of an array of sample wells 19 provided in the sample plate. Each sample well 19 includes 10 pockets, recesses or apertures 21 disposed at the bottom or base of the sample well 19. In use, the reagent beads or microspheres are preferably inserted into each pocket, recess or aperture 21 of the sample well 19 and the reagent beads or microspheres are preferably secured in the pocket, recess or aperture 21 by means of a tapered and restricted diameter of the aperture.

Figure 7B shows the bottom of the sample well 19 in more detail and shows a plurality of pockets, recesses or apertures 21 provided in the bottom of the sample well 19 each arranged and adapted to receive a reagent bead or microsphere. Each pocket, recess or aperture 21 provided in the bottom of the sample well 19 preferably also includes a countersink or region at the entrance to each tapered aperture. According to a preferred embodiment, a single reagent bead or microsphere is dispensed and inserted into each well, recess or aperture 21.

Figure 7C shows in further detail a reagent bead or microsphere 20A arranged and securely positioned in a pocket, recess or aperture 21 provided in the base of the sample well 19 according to the second main embodiment of the invention. The reagent bead or microsphere 20A is secured within the pocket, recess or aperture 21 and the upper surface of the reagent bead or microsphere 20A20A is located or positioned about 0.3mm below the surface of the bottom of the well when secured or positioned within the pocket, recess or aperture 21. Thus, according to a preferred embodiment, the reagent beads or microspheres 20A positioned and secured in the pocket, recess or aperture 21 provided in the bottom of the sample well 19 preferably do not protrude above the entrance or surface of the pocket, recess or aperture 21 and thus preferably do not protrude above the bottom surface of the sample well 19. However, less effective embodiments may be desired in which one or more reagent beads or microspheres 20A positioned in one or more pockets, recesses or apertures 21 provided in the bottom of the sample well 19 may be positioned in a relatively shallow pocket, recess or aperture 21 or may be positioned in one or more pockets, recesses or apertures 21 having a taper such that the reagent beads or microspheres protrude slightly above the entrance or surface of the pocket, recess or aperture 21 and thus above the bottom surface of the sample well 19 when the reagent beads or microspheres 20A are securely positioned in the pocket, recess or aperture 21.

The reagent beads or microspheres are preferably dispensed into a pocket, recess or aperture 21 provided in the bottom of the sample well 19 by means of a reagent bead or microsphere dispenser 22 which will now be described with reference to figures 8A, 8B and 9. A preferred reagent bead or microsphere dispenser 22 according to the second main embodiment is shown in fig. 8A and preferably comprises an upper cover 23, a syringe body 24 and a barrel 25 protruding from a lower region of the syringe body 24.

Fig. 8B shows a cross-sectional view of the reagent bead or microsphere dispenser 22 and shows that the reagent bead or microsphere dispenser further includes a plunger guide 26 preferably positioned within the body of the syringe body 24 according to a preferred embodiment. Plunger guide 26 preferably includes threads on the outer surface of the upper portion of plunger guide 26. The inner surface of the upper portion of the syringe body 24 preferably includes complementary threads that engage with threads provided on the outer surface of the upper portion of the plunger guide 26 to enable the plunger guide 26 to be securely fastened or screwed to the syringe body 24 in use. The inner surface of the cap 23 preferably also includes threads and the cap 23 preferably also threads onto the upper portion of the plunger guide 26.

Plunger 27 is preferably positioned within plunger guide 26 and plunger 27 may be depressed by actuating an actuator or plunger boss 28, plunger boss 28 being positioned in the aperture defined by plunger guide 26 above plunger 27. An actuation spring (not shown) is provided between the actuator or plunger boss 28 such that when the actuator or plunger boss 28 is depressed, force is transferred to the plunger 27 via the actuation spring, causing the plunger 27 to be depressed. A return spring (not shown) is preferably provided between the bottom of the plunger guide 26 and the plunger 27 so that both the plunger 27 and the actuator or plunger boss 28 preferably return to the upper position when the actuator or plunger boss 28 is no longer depressed.

Figure 9 shows an exploded view of a reagent bead or microsphere dispenser 22 according to the second main embodiment and as shown and described above with reference to figures 8A and 8B. Fig. 9 also shows that a silicone element 30 is preferably provided in the upper part of the cylinder 25. In use, the reagent beads or microspheres within the syringe body 24 are preferably leaked or directed through a helical path formed in the bottom region of the syringe body 24 such that the reagent beads or microspheres are arranged in a single row or in series at the bottom of the syringe body 24. A single row or tandem of reagent beads or microspheres is introduced into the chamber preferably disposed immediately above the barrel 25 and below the plunger guide 26. The cavity is formed and arranged to receive a single reagent bead or microsphere positioned in the bore below the plunger 27 and above the barrel 25. When plunger 27 is depressed, plunger 27 preferably pushes individual reagent beads or microspheres 20A located in the chamber in a downward direction. This reagent bead or microsphere 20A is preferably forced through the silicone element 30 by a plunger 27. According to a preferred embodiment, the plunger 27 preferably continues to push or urge the reagent bead or microsphere 20A through the barrel 25 and into the well bore, recess or aperture 21 of the sample well 19 preferably located immediately below the barrel 25 of the reagent bead or microsphere dispenser 22. The silicone element 30 preferably prevents accidental release of the reagent beads or microspheres from the chamber of the reagent bead or microsphere dispenser 22 into the barrel 25 of the syringe body 24.

The bottom of the syringe body 24 preferably has a helical shape and acts to direct or guide the reagent beads or microspheres towards a cavity disposed in the lower portion of the syringe body 24. The chamber is preferably arranged so that only one reagent bead or microsphere is located above the silicone element 30 at any one time. A cavity is formed in the bore through which plunger 27 travels and depression of plunger 27 preferably causes reagent beads or microspheres located in the cavity to be pushed through silicone member 30 and into barrel 25.

A vibration mechanism is optionally provided and may be arranged to act on the outside of the syringe body 24 to ensure that the reagent beads or microspheres move down through the syringe body 24 to the bottom of the syringe body 24 and are aligned in line or series ready to enter the chamber.

The reagent beads or microspheres may be prepackaged or preloaded into the syringe body 24 by, for example, the kit manufacturer or other supplier. Alternatively, the end user may load the syringe body 24 with reagent beads or microspheres.

A microarray spotter or automated device according to a second main embodiment will now be described with reference to fig. 10. As shown in fig. 10, a plurality of syringe bodies 37 may be loaded onto a tray or package 36, and the tray or package 36 is then preferably automatically loaded into a microarray spotter or automated device. A tray or package 36 comprising a plurality of syringe bodies 37 may be moved by a three-axis translation mechanism or robotic arm to a reagent bead or microsphere dispensing work area of a microarray spotter or automated apparatus.

The microarray spotter or automated apparatus preferably comprises a three-axis translation mechanism, which preferably comprises a first translation stage comprising a guide 31, along which guide 31 a first arm 32 is translatable in a first (x) horizontal direction. A second translation stage is preferably provided which includes a mounting block 33, the mounting block 33 preferably surrounding or encircling the first arm 32. The mounting block 33 is translatable in a second (y) horizontal direction (which is preferably orthogonal to the first (x) horizontal direction) and is movable back and forth along the first arm 32. A third translation stage is preferably provided which preferably includes a body or syringe drive mechanism 34 which preferably houses a linear actuator (not shown). The body or syringe drive mechanism 34 is preferably slidably mounted on the mounting block 33 and is liftable in the vertical (z) direction.

The three-axis translation mechanism preferably also includes a telescoping arm 35 preferably extending from the mounting block 33. The tri-axial translation mechanism is preferably programmed to select and pick up reagent bead or microsphere dispensers 22, 37 from a tray or package 36 comprising a plurality of reagent bead or microsphere dispensers 22, 37. The body or syringe drive mechanism 34 comprises a tapered sleeve resiliently mounted within a tubular housing. The cannula is arranged to engage with a tapered portion provided on the syringe cap 23 of the reagent bead or microsphere dispenser 22, 37. When the reagent bead or microsphere dispenser 22, 37 is located in the tray or package 36, the cannula may be lowered onto the syringe cap 23 of the reagent bead or microsphere dispenser 22, 37, thereby detachably securing the reagent bead or microsphere dispenser 22, 37 to the body or syringe drive mechanism 34. The body or syringe drive mechanism 34 and attached reagent bead or microsphere dispenser 22, 37 may then be raised to such a height that the retractable arm 35 (which is initially retracted within the body of the mounting block 33) can be extended. The reagent bead or microsphere dispensers 22, 37 are then lowered by the body or syringe drive mechanism 34 so that the upper portion of the syringe body 24 is secured by the retractable arm 35. The retractable arm 35 preferably has an aperture with an inner diameter preferably smaller than the outermost diameter of the rim of the upper portion of the syringe body 24.

According to a preferred embodiment, each reagent bead or microsphere dispenser 22, 37 preferably comprises a plurality of identical reagent beads or microspheres. According to one embodiment, up to 15 separate reagent bead or microsphere dispensers 22, 37 may be loaded or disposed in a single tray or package 36 and each reagent bead or microsphere dispenser 22, 37 may have a capacity of up to about 2000 reagent beads or microspheres.

According to a preferred embodiment, the syringe drive mechanism 34 is arranged to pick up the reagent bead or microsphere dispenser 22, 37 from the tray or package 36 and will position and lower the barrel 25 of the reagent bead or microsphere dispenser 22, 37 such that it is immediately above the desired reagent bead or microsphere pocket or recess 21 provided in the sample well 19 of the sample plate. The syringe drive mechanism 34 is then preferably actuated so that the actuator or plunger boss 28 of the reagent bead or microsphere dispenser 22, 37 is depressed, which in turn causes the plunger 27 to push the reagent bead or microsphere 20A from the chamber through the silicone member 30, through the barrel 25 and into the desired reagent bead or microsphere pocket or recess 21 of the sample well 19. The syringe drive mechanism 34 is preferably arranged to depress the actuator boss 28 and plunger 27 with a desired amount of force, rather than move the actuator or plunger boss 28 and plunger 27 to a certain vertical position. Thus, the reagent beads or microspheres 20A are preferably pressed tightly and all the way into the reagent bead or microsphere pockets or recesses 21 of the sample well 19 with a constant amount of force.

Fig. 11 shows in more detail the reagent bead or microsphere dispenser picking apparatus or syringe drive mechanism 34 during the process of picking up the reagent bead or microsphere dispenser 22. The reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 comprises a sleeve 39 having a tapered lower end arranged to engage with a tapered recess provided in the upper portion of the syringe cap 23 of the reagent bead or microsphere dispenser 22. The sleeve 39 includes a central aperture through which the plunger rod 40 is mounted. The plunger push rod 40 is arranged to be driven up and down by a linear actuator 41 which drives a linear actuator lead screw 42, the linear actuator lead screw 42 raising and lowering the plunger push rod 40.

As shown in FIG. 11, to pick up the reagent bead or microsphere dispenser 22, the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 is lowered onto the reagent bead or microsphere dispenser 22 such that the cannula 39 of the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 engages the syringe cap 23 of the reagent bead or microsphere dispenser 22. As the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 is driven downwardly onto the reagent bead or microsphere dispenser 22, the sleeve 39 is compressed and moves upwardly until it is prevented from any further upward movement. The sleeve 39 is preferably driven further downwardly under pressure so that the interlocking tapers of the sleeve 39 and syringe cap 23 preferably engage, causing the reagent bead or microsphere dispenser 22 to be attached to the reagent bead or microsphere dispenser picker device or syringe drive mechanism 34.

The reagent bead or microsphere dispenser 22 shown in FIG. 11 is generally similar to that shown in FIGS. 8A, 8B and 9, except that the partition 29 shown in FIGS. 8B and 9 is replaced by a retaining cover 43 in the embodiment shown in FIG. 11. Fig. 11 also shows the position of an actuation spring 44 disposed between the actuator or plunger boss 28 and the plunger 27 and transmitting the force applied to the actuator or plunger boss 28 to the plunger 27. A return spring 45 is also shown and is disposed between plunger 27 and the base of plunger guide 26 and returns plunger 27 (and thus also actuator or plunger boss 28) to the upper position when actuator or plunger boss 28 is no longer depressed or actuated.

Fig. 12A shows the reagent bead or microsphere dispenser picking apparatus or syringe drive mechanism 34 having picked the reagent bead or microsphere dispenser 22 and in the process of delivering the reagent bead or microsphere dispenser 22 to a desired location. Once the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 has been engaged with the reagent bead or microsphere dispenser 22, the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 is raised so that the cannula 39 is no longer pressurized. The sleeve 39 returns to the down position and the reagent bead or microsphere dispenser 22 comprising the syringe body 24 is locked onto the sleeve 39 by the taper on the sleeve 39 and the syringe cap 23.

FIG. 12B shows the reagent bead or microsphere dispenser 22 in the process of dispensing a reagent bead or microsphere 20A from the reagent bead or microsphere dispenser 22 into a pocket or recess of a sample well (not shown) of a sample plate (not shown). The linear actuator 41 of the reagent bead or microsphere dispenser pick-up device or syringe drive mechanism 34 is preferably actuated and causes the linear actuator lead screw 42 to extend to push the push rod 40 downward. The downward movement of the push rod 40 depresses the actuator or plunger boss 28. The actuator or plunger boss 28 transfers force to the plunger 27 via the actuation spring 44 and preferably does not directly contact the plunger 27. Plunger 27 preferably pushes reagent beads or microspheres 20A from a chamber provided within a central bore in syringe body 24. The reagent beads or microspheres 20A are preferably forced by a plunger 27 through the membrane 30 and down through the barrel 25 and into a recess or well of a sample plate (not shown).

FIG. 13A shows the reagent bead or microsphere dispenser picking apparatus or syringe drive mechanism 34 in the process of discharging the reagent bead or microsphere dispenser 22 from the end of the reagent bead or microsphere dispenser picking apparatus or syringe drive mechanism 34. In this mode of operation, the reagent bead or microsphere dispenser 22 is positioned above the tray or package 36. The linear actuator 41 preferably drives the linear actuator lead screw 42 downward until the plunger 27 is maximally extended. The sleeve 39 also extends to a maximum extent. The linear actuator 41 then preferably continues to apply force to the plunger 27 via the actuator or plunger boss 28, as shown in fig. 13B, with the result that the body of the reagent bead or microsphere dispenser 22 is preferably forced away from the end of the tapered sleeve 39. The reagent bead or microsphere dispenser 22 then preferably drops back into the reagent bead or microsphere dispenser tray or package 36.

To illustrate the features of an embodiment of the present invention, a test was performed in which a sample plate comprising 9 sample wells 19 was provided. Each sample well 19 comprises 10 holes, recesses or apertures 21 arranged in a circle around a central portion of the sample well 19. Each well, recess or aperture 21 is loaded with a reagent bead or microsphere coated with a different concentration of reagent. 10 beads in the first sample well were covered with reagent at a concentration of 10 micrograms/milliliter (μ g/ml) and 10 beads in the second sample well were covered with reagent at a concentration of 8 micrograms/milliliter. The 10 beads in the third sample well were covered with reagent at a concentration of 4 microgram/ml and the 10 beads in the fourth sample well were covered with reagent at a concentration of 2 microgram/ml. The 10 beads in the fifth sample well were covered with reagent at a concentration of 1 microgram/ml and the 10 beads in the sixth sample well were covered with reagent at a concentration of 0.5 microgram/ml. The 10 beads in the seventh sample well were not covered with reagent, i.e., at a concentration of 0 μ g/ml. The 10 beads in the eighth sample well are covered by reagents at different concentrations and include concentrations of 10, 8, 4, 2, 1, 0.5, 0 and 0 micrograms/ml. The 10 beads in the ninth sample well have the same concentration and are arranged in the same manner as the reagent beads or microspheres in the eighth sample well.

The reagent beads or microspheres were covered with capture antibody including ovine lgG and delivered in bicarbonate buffer including 0.02% kathon (rtm) preservative.

Sample wells 19 of the sample plate were emptied of the preservative in which the reagent beads or microspheres were transported and 400 microliters of 1/1000 diluted donkey anti-ovine lgG conjugate in Triethanolamine Buffered Saline (TBS) conjugate dilution buffer was added to each sample well 19. The sample plate was then incubated at ambient temperature and subjected to moderate intensity vibration for 45 minutes. Any unbound conjugate is then aspirated from the sample well 19 using a single channel wash head of a microarray spotting device (DS2(RTM), available from Dynex technologies). Once any unbound conjugate has been aspirated from the sample wells 19, 500 microliters of 1/20 diluted triethanolamine buffered saline wash is immediately added to each sample well 19. The process of washing liquid then being sucked out of the sample well 19 and washing and sucking washing liquid out of the sample well 19 is repeated two more times. After the third washing step, comprising aspiration of the washing liquid, has been completed, 300 microliters of luminol (chemiluminescent label) is then added to each sample well 19 immediately. The sample plate is then incubated in the dark at ambient temperature while being subjected to moderate intensity vibration for 15 minutes. The sample plate is then immediately transferred to the reading chamber.

The camera is set to an exposure time of 6 minutes and 30 seconds with a gain of 20. Images were taken at 22 and 29 minutes after the luminol had been added. The camera exposure time was then changed to 8 minutes 37 seconds. Images were further taken at 38, 47, 56 and 65 minutes after the addition of luminol. Image analysis showed that the maximum observed signal intensity consistent with the luminescence ammonia decay curve was obtained 15-22 minutes after the addition of luminescence ammonia.

According to a preferred embodiment, once the reagent beads or microspheres have been dispensed into the wells, recesses or apertures of the sample plate or the reagent bead receiving chamber, the following steps may be performed. First, a sample fluid may be added to one or more sample wells of a sample plate. The sample fluid may include one or more analytes, such as a particular antigen that can react with a reagent coated on one or more reagent beads or microspheres. The reagent beads or microspheres are preferably covered with a specific capture antibody.

Once the sample fluid has been added to the sample well, the sample plate is then preferably subjected to an incubation step. After the sample plate has been subjected to an incubation step such that an antigen-antibody composition is formed, the sample plate is then preferably subjected to one or more washing and aspiration steps to remove any unbound sample fluid and to remove any washing liquid. An enzyme conjugate is then added which will bind to the antigen portion of the antigen-antibody composition that has formed but which will not bind to the antibody or antibody portion of the antigen-antibody composition. The sample plate is then incubated before being subjected to one or more washing and aspiration steps. Once the sample plate has been subjected to one or more washing and aspiration steps, luminol (or another visualising agent) is preferably added. The sample plate is then preferably aspirated to remove any excess luminol (or other visualising agent). Luminol (or other visualization agent) will break down upon contact with an enzyme attached to the antigenic portion of the antigen-antibody composition, causing a particular color to be produced. In the final stage, the sample plate is analyzed and preferably end-point determination is performed.

A particularly preferred embodiment of the present invention is shown in fig. 14A and 14B and will be described in more detail below. Fig. 14A shows 9 sample plates loaded into the plate frame. Each sample plate shown in fig. 14A includes one 6 x 1 sample well. The sample plate can be removably loaded into the plate frame. Each of the 9 sample plates or strips comprises 6 sample wells and each sample well preferably comprises 10 tapered apertures arranged in use to receive reagent beads. The reagent beads are preferably loaded into the tapered aperture so that the reagent beads do not protrude above the base of the sample well. Fig. 14B shows the plate frame into which the sample plate can be loaded in more detail.

Figure 15A shows one 6 sample well in more detail. According to a preferred embodiment, the sample wells in the strip can be separated or disconnected. According to one embodiment, the sample plate or strip can be separated or divided into individual sample wells. Figure 15B shows a strip of 6 sample wells being loaded into the plate frame.

Figure 16A shows a single sample well being loaded into the plate frame. (which has been isolated from one sample well). The sample well preferably comprises a female portion which is preferably arranged to engage or interlock with a male portion preferably provided on the base of the plate frame. The sample plate or sample strip is preferably arranged to be securely fastened and fixed to the plate frame when loaded onto the plate frame.

Figure 16B shows the two sample wells connected by the disconnect structure 47 in more detail. The break-off structure 47 preferably allows a user to separate adjacent sample wells. According to one embodiment, the sample wells may be separated from each other but may still be arranged adjacent to each other on the plate frame while not interfering with each other. The breaking structure 47 preferably comprises one, two or more than two breaking points 46. According to one embodiment, the connection 47 between two sample wells may be disconnected from the sample wells at the first breakpoint 46. The connector 47 can then be disconnected or removed from the sample well to which it is attached by disconnecting the connector 47 from the sample well at the second breakpoint 46.

Fig. 16C shows a sample well with an end break-off structure 48. The end break-off structure 48 allows the end wells to be used individually in the plate frame without disturbing another sample well. The end break-off structure 48 is provided to facilitate grasping by a user to remove one sample well or a single sample well from the plate frame.

Fig. 16D shows a sample well with ID and orientation tag 49. Label 49 allows for printing an identifier onto label 49 or otherwise attaching to label 49. The identifier may comprise a 2D or 3D barcode and/or text readable by a person. Labels 49 preferably assist the user in orienting sample wells by aligning with structures in the plate frame and/or on other sample wells when using a single sample well.

Figure 17A shows the underside of one sample well and shows that each sample well comprises a bore or recess into which 10 reagent beads are preferably inserted in use according to a preferred embodiment. The base or underside of each sample well preferably further comprises a concave portion which is preferably arranged to cooperate, in use, with a convex portion provided in the base of the plate frame.

Figure 17B shows in more detail the female alignment and retaining members 50 that help align one sample well with the plate frame. Fig. 17C shows a corresponding male alignment and retention member 51 preferably provided in the base of the panel frame. According to one embodiment, male portion 51 may comprise a plurality of flexible protrusions that preferably deform inwardly when a sample well is positioned on male portion 51. The projections on the plate frame preferably move or come closer together, ensuring that the sample wells remain in place without undue force being applied to mount or secure the sample wells to and/or dismount the sample wells from the plate frame.

Fig. 18 shows a cross-sectional view of one sample well and shows that the sample well preferably has a plurality of tapered apertures 52 according to a preferred embodiment. The tapered bore 52 preferably serves as a cavity into which reagent beads may be inserted in use. The angle of the taper is preferably 6.0 °.

Although the various embodiments described above have focused on reagent beads coated with biomolecules for use in immunoassay or ELISA procedures, the invention is equally applicable to reagent beads that include or are otherwise coated with nucleic acid sequences and that serve as hybridization probes for detecting DNA or RNA sequences complementary to those provided on the reagent beads. As will be appreciated by those skilled in the art, the hybridization probe will be inert until hybridization, at which point there is a conformational change and the molecular synthesis becomes active and will then emit light under uv light. Thus, all of the various embodiments described above are equally applicable to the use of reagent beads that include or are otherwise covered with DNA or RNA sequences for use as hybridization probes for detecting complementary DNA or RNA sequences.

Although the present invention has been described with reference to preferred embodiments, workers skilled in the art will recognize that changes may be made in form and detail without departing from the scope of the invention, as set forth in the appended claims.

Claims (11)

1. A combination comprising a sample plate comprising one or more sample wells and reagent beads or microspheres, wherein one or more of the sample wells comprise:

a base;

a plurality of voids or depressions disposed in the base, wherein the plurality of voids or depressions each comprise an aperture having a tapered section;

wherein, in use, a reagent bead or microsphere is inserted or positioned within each of said apertures; and

the opening to each aperture and/or the cross-sectional shape of each aperture is circular; and

wherein each said reagent bead or microsphere is substantially retained or secured within one of said apertures by an interference or friction fit with said tapered section of said aperture, and wherein said reagent bead or microsphere forms a substantially fluid tight seal with said tapered section of said aperture.

2. The combination of claim 1, wherein one or more of the sample wells are interconnected to one or more other sample wells by one or more frangible regions or joints, such that the sample plate can be separated into a plurality of smaller sample plates by a user.

3. The combination of claim 1, wherein the sample plate comprises an immunoassay sample plate.

4. The combination of claim 1, wherein at least some or substantially all of the reagent beads or microspheres carry, comprise or are covered with a reagent, wherein the reagent is arranged and adapted for determination of an analyte of interest in a sample liquid.

5. The combination of claim 1, wherein the sample plate comprises hybridization probes for detecting the presence of a complementary DNA or RNA sample.

6. The combination of claim 5, wherein at least some or substantially all of the reagent beads or microspheres carry, comprise or are covered with a nucleic acid probe, wherein the nucleic acid probe is arranged and adapted to hybridize to single stranded nucleic acid, DNA or RNA.

7. A method of analyzing a plurality of analytes in a sample using a combination comprising a sample plate and reagent beads or microspheres, comprising:

providing a combination according to claim 1; and

adding a sample to the sample well.

8. A method for determining one or more analytes of interest in a sample, comprising:

inserting a plurality of reagent beads or microspheres into a plurality of pockets or recesses of one or more sample wells of a sample plate, wherein the sample wells comprise a base and the plurality of pockets or recesses disposed in the base, wherein the plurality of pockets or recesses each comprise a bore having a tapered section;

the method is characterized in that:

the opening to each aperture and/or the cross-sectional shape of each aperture is circular; and

wherein each said reagent bead or microsphere is substantially retained or secured within one of said apertures by an interference or friction fit with said tapered section of said aperture, and wherein said reagent bead or microsphere forms a substantially fluid tight seal with said tapered section of said aperture.

9. A kit for performing an enzyme-linked immunosorbent assay (ELISA) procedure, comprising:

one or more sample plates comprising one or more sample wells, wherein one or more of the sample wells comprise a base and a plurality of pockets or recesses disposed in the base, wherein the plurality of pockets or recesses each comprise an aperture having a tapered section; and

a plurality of reagent beads or microspheres coated with a reagent comprising an antibody, an antigen or another biomolecule;

the method is characterized in that:

the opening to each aperture and/or the cross-sectional shape of each aperture is circular; and

wherein each said reagent bead or microsphere is substantially retained or secured within one of said apertures by an interference or friction fit with said tapered section of said aperture, in use, and wherein said reagent bead or microsphere forms a substantially fluid tight seal with said tapered section of said aperture, in use.

10. A kit for performing a nucleic acid detection process, comprising:

one or more sample plates comprising one or more sample wells, wherein one or more of the sample wells comprise a base and a plurality of pockets or recesses disposed in the base, wherein the plurality of pockets or recesses each comprise an aperture having a tapered section; and

a plurality of reagent beads or microspheres coated with DNA or RNA sequences;

the method is characterized in that:

the opening to each aperture and/or the cross-sectional shape of each aperture is circular; and

wherein each said reagent bead or microsphere is substantially retained or secured within one of said apertures by an interference or friction fit with said tapered section of said aperture, in use, and wherein said reagent bead or microsphere forms a substantially fluid tight seal with said tapered section of said aperture, in use.

11. A method of manufacturing a combination comprising a sample plate and reagent beads or microspheres, comprising:

providing a sample plate comprising one or more sample wells each having a base;

forming a plurality of pockets or recesses in the one or more bases, wherein the plurality of pockets or recesses are arranged and adapted to receive, in use, reagent beads or microspheres, the plurality of pockets or recesses each comprising a bore having a tapered section, and wherein the opening to each bore and/or the cross-sectional shape of each bore is circular; and

inserting or positioning a reagent bead or microsphere within each of said apertures;

wherein each said reagent bead or microsphere is substantially retained or secured within one of said apertures by an interference or friction fit with said tapered section of said aperture, and wherein said reagent bead or microsphere forms a substantially fluid tight seal with said tapered section of said aperture.