DE1948191U - DEVICE FOR CARRYING OUT ELECTROPHORETIC EXAMINATIONS. - Google Patents

Description

RA. ^ f 78 636 * 9/10/66 ^

10 256 gm

H 55 840/42 1 Gbm

Hyland Laboratories, 4501 Colorado Boulevard, Los Angeles, California (USA)

Device for performing electrophoretic

Investigations

The innovation relates to devices in which mixtures due to the different travel speeds their components can be separated by a transport medium in an electric field, and in particular to a new and improved, single-use, plastic electrophoresis test device for sorting out or separating particles in a buffer arrangement to which direct current is applied is.

Numerous electrophoresis test devices of numerous sizes are known to measure components of a mixture

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to be able to sort out something, whereby each material has its own special charge, which differs from that of others Materials differ in size and type (sign). The electrophoretic separation and subsequent agglomeration of materials or particles usually takes place within a supporting medium that serves as a buffer, wherein the particles of a sample to be examined, which have a greater charge, at a greater speed and also migrate further in a certain time than particles with a lower charge. After the particles have grouped together, the ^ ptical determination (visual identification) of the material in the to be examined sample carried out in a known manner, for example by colorimetric analysis.

Up until now, expensive and relatively complex devices were required to carry out these methods, which required operation by highly qualified technicians and also a great deal of time for detailed monitoring of the preparation, operation and subsequent cleaning of the test device. All of these steps, together with the high installation costs of the necessary equipment, hindered the possible application of electrophoresis as a suitable test method, particularly in the clinical field, on the one hand
may have been accepted for a variety of reasons, both economic and other reasons.

It is always in the field of application mentioned above have become more useful, for example, to determine the presence of the individual isozymes of lactate dehydrogenase to be able to. It should be understood, however, that this is only one use case is, in which the electrophoretic separation can be carried out and here only as an exemplary embodiment is called.

Enable inexpensive, single-use, self-contained or independent electrophoresis devices It also allows relatively small clinics to apply diagnostic procedures that were previously only available by large commercial ones Laboratories or institutes could be carried out. In addition, the previously used electrophoresis devices are designed so that each structure requires the gathering and assembling of more numerous Requires individual parts of an expensive and sometimes space-consuming equipment. Around To be able to carry out more than one examination at the same time, a double device was previously required, whereby the installation costs are increased and the possibility of errors due to incorrect assembly and inaccurate cleaning increase after use.

The present innovation, on the other hand, creates a consumable, single-use vessel-like device for clinical purposes. By the properties of this

Device it is possible for the operator under normal circumstances to work with much fewer individual parts and further relieves you of the assembly and cleaning problems described above that occur with Reuse of every piece of laboratory equipment. Many of the problems described above are avoided by when you have all the necessary components, the required material and all the facilities in a single one Houses packaging that od a plurality of a transport medium receiving lids. The like. Comprises.

It is therefore the main task of the furnace to provide a new and improved, relatively cheap, one-time use only To create electrophoresis testing device.

Furthermore, it is the task of the innovation to provide devices or possibilities for electrophoretic investigations create, taking all the necessary test components for electrophoretic Secretion of particles contained in a single container.

It is also an object of the innovation to provide a new and improved device for holding a pre-cast To create gels through which an electrical current to generate the particle migration can be passed can.

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Furthermore, it is the task of inflation, new and novel To create facilities to place samples to be examined on a suitable gel support medium, so that a electrophoretically ^ particle migration takes place.

Finally, it is the object of the innovation to create an improved device for an electrical Pass current through a buffer in a gel medium within an electrophoresis test device be able.

Accordingly, one characteristic of inflation is that that a compact portable electrophoresis tester was created, which consists of plastic or other plastic material and a basic chamber with in it has arranged removable contact liquid or buffer containers, electrodes and a lid, which holds a gel support medium as a connection between the buffer containers.

Another feature of the innovation was a multi-compartment, one-piece design Electrophoresis chamber created in which two spaced apart / arranged buffer containers each with a buffer are provided, which are in contact with a gel medium, which od in a plastic lid. The like. Arranged is to be able to apply direct current to the system,

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rather should flow through the gel medium.

Another feature of the innovation was a One-piece electrophoresis chamber created with several compartments spaced from each other Two buffer containers containing electrodes with a wick are provided to keep a buffer mixture in contact to hold with a gel medium which has been poured into a plastic lid and solidifies there so that direct current can be applied to the system and pass through the gel medium.

Another feature of the control is to have a Plastic lid or a corresponding cap made of plastic or other suitable material for an electrophoresis chamber to create, which contains a gel support medium and is designed so that on him at least one further lid can be attached so that a large number of lids compact with a single basic chamber can be sent assembled.

Another feature of the innovation is to create a template or template with the indentations can be generated for samples to be examined at a predetermined distance in the solid gel medium.

Another feature of the innovation is to provide sheet-like electrodes that are designed in this way

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are that they practically fit in two buffer containers and can be connected to a power source.

The above-mentioned purposes and objectives and features of the innovation are based on the following description explained in more detail in the drawing »

In the drawing are two exemplary embodiments of the electrophoresis device shown according to the innovation, namely shows

Mg. 1 is a plan view of an electrophoresis container,

Mg. 2 is a plan view of the container from Mg. 1 on an enlarged scale without the lid,

Mg. 3 a longitudinal section along line 3-3 from Mg. 1,

Mg. 4 is a diagrammatic view of a cover and a template matching it,

Mg. 5 a partial section along one of the lines 5-5 from Mg. 1,

Fig. 6 is a plan view of an electrophoresis container according to another embodiment the innovation,

Mg »7 a plan view of the container made of Mg. 6 on an enlarged scale without the lid,

Mg. 8 a longitudinal section along line 8-8 from FIG. 6

Pig. Figure 9 is a side view of the Pig container. 6 and

Pig. 10 a partial section as in Pig. 5 »from which the

Position of a foil electrode in the operating position can be seen.

According to Pig. 1, the electrophoresis apparatus has a container 10 made of a substantially rectangular shape Electrophoresis chamber 12 with side walls 14 and end walls 16 which are attached to a base plate 18, consists. Each of the walls 14 and 16 is made thinner in the region of its upper edge, so that an L-shaped shoulder 15 for placing a lid on each wall 14 and 16 is provided. The chamber 12 is closed with a lid 20 which rests on its upper side. The in detail in the Pig. 3 illustrated cover 20 has reduced external dimensions at its top, so that an outer shoulder 22 is formed, onto which further identical covers can be attached one over the other, so that they can be transported in a compact unit with the electrophoresis container 10. Everyone Lid 20 is also provided with a similarly formed inner shoulder 23, so that it can be on the walls 14 and 16 of the chamber 12 can be attached. The upper ends of the walls 14 and 16 have together with the inner shoulder 23 of lid 20 mating complementary contours, such as from Pig. 3 and 5 to be recognized

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In the plan view of the container shown in FIG 10, the lid 20 is removed from the chamber 12. In the chamber 12 are two rectangular buffer containers and 26 arranged. These containers 24 and 26 are open at their top and parallel to the end walls 16 of the Chamber 12 arranged. On the base plate 18 are vertically upwardly extending supports 28 as a support for the containers 24 and 26 are arranged, which hold these containers in the chamber 12 at a certain height. Besides that keep the supports 28, the containers 24 and 26 in the chamber 12 at a certain distance from each other until they are lifted out of the chamber 12. Recesses 32 are provided on the outside of the side walls 14, In which Nobben 33 or the like. Can intervene, which are arranged on the inside of the cover 20 and hold this firmly on the chamber 12 during electrophoresis. The containers 24 and 26 can also be carried outwards above, not shown hanging eyelets or the like. Be provided to them in the chamber 12 on the walls hang up.

A foil electrode 34 with suitable electrical conductivity properties is mounted in each buffer container 24 and 26 and is adapted to be conveniently inserted into the container 24 and 26, as shown in Figs. 2 and 3, fits therein. Each foil electrode 34 has an outward one protruding tongue 36 which protrude from the chamber 12

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can. The tongues 36 are connected to a suitable direct current source 38 via a switch 39. The tongues 36 are in Pig. 2 in their working position to the outside shown unfolded. It will be understood that while foil electrodes are shown, others are known Electrodes such as chromium nickel or platinum wire (as shown in Fig.) Can be used. Become common two electrodes are supplied with each lid 20 of a container 10, all of which are in the free part before use the chamber 12 can lie between the buffer containers. The electrodes can, however, also be placed in the buffer container 24 and 26 coated, evaporated or imprinted.

When the device is assembled for shipping, the tongues 36 of the foil electrodes, as in FIG. 3 shown, be folded onto the top of a sponge-like wick 40, which is in each of the buffer containers 24 and 26 of the container 10 is arranged. The sponge wicks 40 of suitable porosity hold and direct a buffer solution or contact liquid which is poured into containers 24 and 26 prior to the start of electrophoresis a contact or buffer connection between the containers 24 and 26 and the gel support medium discussed in connection with FIG. The sponge wicks 40 are shown to fit into containers 24 and 26 and correspond substantially to the internal dimensions

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this container when they are soaked with buffer solution to saturation. You are touching the gel inside of the lid 20 when the chamber 12 is closed by the same.

In Pig. 3, a cover 20 is placed on the chamber 12 and closes the same so that the liquid contained in it cannot evaporate or evaporate. The lid 20 is also designed to hold a suitable gel medium 46 can receive and hold in a gel reservoir 42. The one provided on the inside of the cover 20 Gel reservoir 42 is delimited by vertical side walls 44 of cover 20. It will be understood that the gel medium can be pre-cast and packaged in this form by the manufacturer and supplied with the lid 20; on the other hand but it can also be poured in by the consumer. When the lid 20 is in the closed position on the Chamber 12 is located, the gel medium is in the end regions of the lid with electrical contact on the sponge wick 40. The lid, the buffer container and the Sponge wicks are chosen in terms of their dimensions so that the gel touches the sponges sufficiently, but they do do not compress too much. On the inside of the cover 20 are vertical Halteste.ge 48 for the gel arranged, which are spaced from the side walls of the lid and holding the semi-solid Medium 46 supported within the lid 20 when the same upside down and in its working position on the Chamber 12 of the device is attached.

In! Pig. 3 is a second cover 50 with interrupted Lines shown to show how two identical lids can be plugged together. The covers 20 and 50 are designed the same and can thus be a Form a compact package for shipping or storage. This way you can put multiple lids on a single one Chamber 12 are attached.

Fig. 4 shows a cover 20 in a position in which a gel insert is poured or inserted into it can be. The lid 20 is provided on the inside of its walls 44 with lobben 33, which are made with the walls consist of one piece and can engage in the recesses 32 on the outside of the chamber 12 when the lid is attached to the chamber. In this way a relatively firm and tight fit is achieved, to hold the gel medium 46 on the sponges 40 and also to allow liquid to evaporate during electrophoresis to prevent from the container.

In Fig. 4, a template 52 is shown which is used to create wells for receiving to be examined To be able to attach samples in the gel support medium 46 located on the inside of the container lid. The template 52 is provided with a plurality of openings 53 through which tools for drilling the Depressions can pass through.

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In FIG. 5, part of the cover 20 slipped onto the chamber 12 is shown in an enlarged detail. The foil electrode 34 is folded onto the top of the sponge wick 40 so that the foil tongue 36 does not move is in the operating position. The foil electrode 34 lines the inner side wall and the bottom of the subject Buffer container 24 and 26 and can be folded outward so that they are between the lower surface the side wall 44 of a lid 20 and the upper end of the end wall of the chamber 12 (as shown in Fig. 10) can pass through. When the foil electrode 34 is unfolded into its operating position, the entire upper side comes up of the sponge wick 40 with the buffered gel medium in touch,

Fig. 6 is a plan view of a modified electrophoresis test apparatus and shows a lid 20 resting on a chamber 56 having side walls 58 and end walls 60 is arranged. The interior of the chamber 56 is provided with partition walls 62 and 64 (FIGS. 7 and 8), which consist of one piece with it and form two buffer containers 66 and 68 with the walls 58 and 60, which are arranged in the longitudinal direction at a distance from one another in the chamber 56. The walls 62 and 64 are perpendicular like the walls 58 and 60 of the chamber 56. In the buffer containers 66 and 68 are sponges and sponge wicks, respectively 40 arranged to lead a buffer solution to a gel medium which is held in the lid 20. By public

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openings 72 in the side walls 58 is a wire electrode 70 inserted into each buffer container. The lid 20, which closes positively on the upper edge the side walls 58 and 60 of the chamber 56 can be attached, is similar to the lid shown in FIG educated.

In Pig. 8, the cover 20 is shown slipped onto the chamber 56. In addition, the partition walls 62 are shown in FIG and 64, which divide the chamber 56 into three compartments, namely the buffer containers 66 and 68 and an empty chamber 74- between these two containers,

fig. 9 is a side view of the electrophoresis apparatus; wherein on the top of the lid 20 an ice pack 76 is applied as a coolant during is used for electrophoresis.

Pig. 10 is a partial section similar to Pig. 5 and shows the foil electrode 34 unfolded in its working position, between the mutually complementary surfaces of the end walls of the chamber 56 and the Lid 20 is passed.

TJm materials electrophoretically due to different particle migration in an electrically conductive buffer system, through which an electric current flows when using the movable test devices described above.

To be able to separate directions, the sample to be examined, which proteins, peptides and amino acids or Contains similar, electrophoretically separated materials, arranged in a buffered support medium. The carrier medium described and shown here is a gel, although other media such as filter paper or cellulose acetate can also be used. The gel pre-cast by the supplier or by the user of the device can be manufactured and cast is in a lid 20 of a test container of the type described above Art included. The sample or samples to be examined are in contact at certain points arranged with the solid gel. If it is desired to place the sample in a specific place in the gel, A template 52 can be used to determine the location of depressions or recesses in the gel. These recesses are made by suitable facilities generated, which can be passed through the openings 53 of the template 52.

The buffered gel is, as described above in connection with Pig. 5 discussed, on the inside of the cover 20 through its walls 44 and also through the retaining webs 48 held. Therefore, when the sample to be examined has wetted the gel, the lid can be turned over and opened the chamber 12 can be attached. Since the buffer container and 26 or «66 and 68 are provided with wicks 40, which

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with their upper side practically parallel and at the level of the If the upper edges of the chamber 12 or 56 are located, the gel medium remains in electrical contact with ■ during operation those soaked with contact or buffer fluid Sponge wicks 40 * Before the lid is put on, Contact or buffer solution poured over wicks 40 in containers 24 and 26 and 66 and 68, respectively. As the gel medium is pre-buffered in the cover 20, there is a continuous flow when the cover is brought into its working position Current path of contact or buffer liquid from one container 24 or 66 through the gel medium 46 in the second buffer container 26 or 68, so that after closing the switch 39 a closed circuit given is.

In order to start the electrophoresis, direct current is passed through the device. This is achieved in that one within the buffer container 24 and 26 arranged polar electrodes 34 connected to the power source. It goes without saying that by applying electricity to the electrodes 34 a migration of buffer salt ions is caused by the buffer liquid and wander the gel. During operation, the buffer liquid and the gel are indicative of the applied direct current Heat Generated * This heat, if insufficiently dissipated, would constitute the proper puncture of the device affect. According to FIG. 9, an ice pack 76 is therefore attached to the top of the device.

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arranges which is to dissipate the heat generated in the chamber. It is understood, however, that any other suitable Cooling device can be used as a heat exchanger for the same purpose. The coolant can also be inside of the container in the free space in the middle of the chamber.

The potential remains applied to the device for a certain period of time, whereupon by means of a suitable Analysis method localizes the electrophoretically separated components of the sample examined and be identified. It will be understood that the direction and rate of molecular or particle migration Punctures of the size of the electric field, the charge of the particles, the pH value of the buffer liquid and other factors such as particle size, viscosity, and the like. Using the phrase "particle" parts with very small dimensions such as molecules of amino acids etc. are referred to.

The innovation provides a relatively inexpensive electrophoresis device that can be used once or consumed created, which has all the necessary boundary conditions for examining samples and is designed so that it forms a compact unit, which contains several gel-containing lids and only one movable Electrophoresis chamber includes.

Protection claims:

Claims (11)

Protection claims; 1. - One-time use electrophoresis test device, ¹ characterized in that it has a plastic chamber (12, 56) with two contact or buffer liquid containers (24, 26 or 66, 68) arranged in it at a distance, as well as devices, in order to hold a buffered support medium (46) between the buffer liquid containers and in electrical contact with the buffer liquid in each container, these devices comprising a largely bowl-shaped plastic cover (20, 50) for attachment to the chamber and electrodes ( 54) protrude in order to allow the passage of electrical current through the buffered medium so that particles or molecules provided with charges can migrate in this medium and then particle group identification or determination is possible. 2. Forrichtung according to claim 1, characterized in that that the buffered medium (46) is a gel, 3. Device according to claim 1 or 2, characterized in that that the plastic chamber (12, 56) with facilities is provided to place a bridge of buffered gel in the bowl-shaped plastic lid (20, 50) 19th to keep, with an outer shoulder (22) on the lid is provided so that identical cover can be slipped onto the same space-saving and thus a Fixed connection between several lids and a single container or a single chamber formed will. 4. Device according to one or more of claims 1 to 3j, characterized in that the chamber (12, 56) is substantially rectangular and open on its top and that the buffer liquid container two spaced apart plastic containers which can be removed from the chamber (12) (24, 26) are. 5. Device according to one or more of claims 1 to 4? characterized in that the removable containers (24 _S 26) are open at their top. 6. Device according to one or more of claims 1 to 3j, characterized in that the chamber (56) is substantially rectangular and open on its top and that the buffer liquid container (66, 68) each have a vertically extending wall (62 or 64), each close to a 20 - the front ends of the chamber are arranged and are made in one piece with the same qxib, and that the walls run perpendicularly in the same direction as the outer walls (58, 60) of the chamber (56). 7. Device according to one or more of claims 1 to 6, characterized in that the electrodes (34) consist of a film or a thin sheet of metal. 8th." Device according to one or more of the claims 1 to 7, characterized in that in the contact or buffer liquid containers (24, 26 or 66, 68) the liquid-absorbing wicks (40) or the like are arranged. 9. Device according to one or more of claims 1 to 8, characterized in that one: with the Lids (20, 50) cooperating template (52) is provided to the samples to be examined to be able to introduce into the support medium (46) made of gel at predetermined points. 10. Device according to one or more of claims 1 to 9), characterized in that each cover (20, 50) has an essentially flat base plate, at the edges of which walls (44) run vertically 21 are arranged and that means for holding the support medium consisting of gel (46) on the Lid base plates are provided to keep the gel in the lid even when the lid is turned upside down or is turned over to bring the gel into electrical contact with the buffer liquid containers (24j 26 or 66, 68) to bring, and that locking devices (32, 33) are provided to the To attach the lid to the chamber (12, 56) or to another lid and the chamber effectively against Seal any vaporized or evaporated liquid from escaping from it during electrophoresis. 11. The device according to claim 10, characterized in that the cover (20, 50) is designed such that that it covers the chamber (12, 56), and that the latch (53) on the vertical walls (44) of the lid are provided. G / MK