CN104677870A - Superminiaturization multi-channel real-time fluorescent spectrum detector - Google Patents
Superminiaturization multi-channel real-time fluorescent spectrum detector Download PDFInfo
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- 238000012360 testing method Methods 0.000 claims description 20
- 150000007523 nucleic acids Chemical class 0.000 claims description 19
- 102000039446 nucleic acids Human genes 0.000 claims description 19
- 108020004707 nucleic acids Proteins 0.000 claims description 19
- 230000003321 amplification Effects 0.000 claims description 18
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 18
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- 230000001678 irradiating effect Effects 0.000 description 1
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Abstract
The invention discloses a superminiaturization multi-channel real-time fluorescent spectrum detector. The superminiaturization multi-channel real-time fluorescent spectrum detector comprises a sample cell, an excitation light source, a fluorescent transmission channel and a multi-spectrum sensor, wherein the fluorescent transmission channel is used for acquiring fluorescent signals formed by emitting light, and the fluorescent signals are effectively output via optical fibers or optical fiber bundles for detection; the multi-spectrum sensor comprises a band pass filter and a photoelectric diode; the band pass filter can be used for spectrum selection of fluorescence with different wavelengths; the photoelectric diode induces the fluorescence intensity of the wavelength corresponding to the band pass filter matched with the photoelectric diode. According to the technical scheme, the multi-spectrum sensor replaces the traditional filter light splitting system, so that the device structure is simplified greatly, and the size of the detecting system is reduced; the energy loss of emitting fluorescence in optical path transmitting is reduced by adopting the fluorescent transmitting channel consisting of optical fibers or optical fiber bundles; compared with the traditional opened type transmitting structure, the superminiaturization multi-channel real-time fluorescent spectrum detector is cleaner, cannot be polluted by dust, and improves the accuracy and flexibility of experiments.
Description
Technical field
The present invention relates to the equipment detected for laboratory or medical treatment, particularly relate to a kind of subminiaturization hyperchannel real-time fluorescence spectrum detection device.
Background technology
At present, in laboratory or medical detection technology, by adding fluorophor in reaction system, fluorescence signal is utilized to accumulate the process of the whole reaction of Real-Time Monitoring, by the qualitative features of the intensity determination reaction system of fluorescence signal, or widely use the method that unknown testing sample carries out quantitative test according to typical curve, wherein the most typical representative is real-time fluorescence quantitative PCR.Specifically, real-time fluorescence quantitative PCR adds fluorophor in PCR reaction system, utilizes fluorescence signal to accumulate the whole PCR process of Real-Time Monitoring, finally by typical curve, unknown template is carried out to the method for quantitative test.Quantitative real time PCR Instrument adds fluoroscopic examination and analytic system on regular-PCR instrument basis.As shown in Figure 1, at the end of each circulation of PCR, the excitation source 101 of instrument launches exciting light through light filter 1, spectroscope 103, refracting telescope 104 and lens 105, finally project in the reactant liquor of amplification pipe 106, after fluorescent material in reactant liquor is subject to exciting of exciting light, produce specific utilizing emitted light, through lens 105, refracting telescope 104, spectroscope 103, light filter 1 and poly-lens 108, finally on photelectric receiver 109, obtain fluorescence signal.Image transmitting, to computer software, demonstrates fluorescence intensity growth curve by analysis afterwards, and the power of fluorescence is measured to the DNA in sample and is directly proportional.
According to fluorescence excitation principle, no probe primer is the exciting light of corresponding different wave length and utilizing emitted light often.Therefore in actual applications, hyperchannel fluorescence detecting system is often needed, i.e. multiple excitation source and multiple utilizing emitted light passage.Tradition is applied to fluoroscopic examination in PCR instrument device and fluorescence spectrophotometer system utilizes a series of independently spectroscope, bandpass optical filter, the combined system of lens combination and photelectric receiver.Such as, the LS32 produced by American I daho company is the fluorescence detecting system of an excitation source and 3 transmission channels.The lightCycler 480 produced by Roche company is the fluorescence detecting system of the corresponding multiple transmission channel of multiple excitation source.Be not difficult to find out from above several conventional fluorescent spectral detection system, traditional real-time fluorescence quantitative PCR instrument ubiquity volume is large, complex structure, the problem of easy pollution, and it is large along with volume, equipment cost is also there is natch too high while complex structure defect, drop into huge problem, general medical science or research laboratory are difficult to bear the problem that this Technical investment etc. is not easy to Technique Popularizing application, and easily to pollute and therefore the problem of sensitivity after the accuracy brought due to complex structure also can hardly be avoided, simultaneously also there is the technical barrier being unfavorable for instrument miniaturization and monommechanization design objectively determining traditional real-time fluorescence quantitative PCR instrument in all these defects.
Summary of the invention
For deficiency of the prior art, the technical matters that the present invention solves is to provide a kind of subminiaturization hyperchannel real-time fluorescence spectrum detection device, use multispectral sensor to instead of traditional optical filter beam splitting system, greatly simplify the complexity of device structure and reduce the volume of detection system.
The technical matters that the present invention solves also is to provide a kind of subminiaturization hyperchannel real-time fluorescence spectrum detection device, adopt the fluorescence transmission channel be made up of optical fiber or fibre bundle, reduce the energy loss of emitting fluorescence in paths, and the transmission mode of this fluorescence signal is cleaner than traditional open transmission structure, be not easy to by contamination by dust, improve accuracy and the sensitivity of experiment.
In order to solve the problems of the technologies described above, on the one hand, embodiments provide a kind of subminiaturization hyperchannel real-time fluorescence spectrum detection device to comprise:
Sample cell, holds needing test sample product and fluorescent material concentration dependent with testing sample in this sample cell;
Excitation source, for the fluorescent material in excited sample pond, fluorescent material be stimulated light source produce excitation after produce utilizing emitted light;
Fluorescence transmission channel, for collecting the fluorescence signal that utilizing emitted light is formed, the fibre bundle that this fluorescence transmission channel comprises optical fiber or is made up of optical fiber, fluorescence signal is effectively exported by optical fiber or fibre bundle and detects;
Multispectral sensor, described multispectral sensor comprises one or more bandpass filter and the photodiode corresponding with described bandpass filter quantity; This multispectral sensor receives and detects the intensity of the fluorescence signal come from the transmission of fluorescence transmission channel, described bandpass filter realizes the optical electivity of different wave length fluorescence, and photodiode induces the fluorescence intensity of the wavelength corresponding to bandpass filter matched with it.
As preferred version of the present invention, a kind of subminiaturization hyperchannel real-time fluorescence spectrum detection device that embodiments of the invention provide comprises the part or all of of following technical characteristic further:
Preferably, the described fluorescence transmission channel fibre bundle that comprises reflective mirror, convergent lens and optical fiber or be made up of optical fiber; Fluorescent material be stimulated light source produce utilizing emitted light direct projection or by after mirror reflection by convergent lens assemble to optical fiber or fibre bundle light entrance end.
Preferably, described fluorescence transmission channel also comprises the mirror array and convergent lens lens arra that are arranged at described sample cell both sides, multiple reflective mirror be arranged in parallel is comprised in mirror array, the reflective surface of this reflective mirror is arranged to the spill matched with the scope of its light collecting zone, described convergent lenses array comprises the convergent lens corresponding with reflective mirror quantity, each reflective mirror and the convergent lens matched form a phosphor collection unit, fluorescence signal is sent to described multispectral sensor via optical fiber or the fibre bundle that is made up of optical fiber after being collected by this phosphor collection unit again.
Preferably, fibre bundle is provided with in described fluorescence transmission channel, the optical fiber that many light entrance ends are arranged as one is included in described fibre bundle, the light entrance end of each optical fiber is corresponding with a phosphor collection unit, and the every root optical fiber in fibre bundle all forms the fluorescence channel that an individual wavelengths is selected.
Preferably, be provided with many simple optical fibers in described fluorescence transmission channel, the light entrance end of each simple optical fiber is corresponding with phosphor collection unit and form the fluorescence channel that an individual wavelengths selects.
Preferably, described multispectral sensor comprises the multiple bandpass filters being integrated in its surface, and the photodiode corresponding with described bandpass filter quantity, and described bandpass filter is fitted the surface of described photodiode; Each bandpass filter and the photodiode matched form a fluorescence intensity detecting unit, for detecting the fluorescence intensity of the wavelength corresponding to bandpass filter that this fluorescence intensity detecting unit is arranged; The light exit end of each optical fiber is corresponding with a fluorescence intensity detecting unit.
Preferably, more than described excitation source is provided with one or two, plural excitation source produces the exciting light of different frequency.More preferably, described excitation source is for being laser diode or LED light source.
On the other hand, the embodiment of the present invention additionally provides a kind of pick-up unit for laboratory or medical treatment, and it comprises the subminiaturization hyperchannel real-time fluorescence spectrum detection device described in such scheme.
Further, the embodiment of the present invention additionally provides a kind of nucleic acid amplification detector, comprise the subminiaturization hyperchannel real-time fluorescence spectrum detection device described in such scheme; Described sample cell is also provided with and realizes the temperature controlled temperature control probe of nucleic acid amplification reaction.
Compared to prior art, technical scheme of the present invention at least has following beneficial effect:
The miniaturized multichannel real-time fluorescence spectrum detection device that the embodiment of the present invention provides, uses multispectral sensor to instead of traditional optical filter beam splitting system, greatly simplify the complexity of device structure and reduces the volume of detection system.Multispectral sensor comprises bandpass filter and photodiode in the inventive solutions, and bandpass filter forms a fluorescence intensity detecting unit with the photodiode matched, utilize the bandpass filter being integrated in multispectral sensor surface to realize the optical electivity of different wave length, the fluorescence intensity of the wavelength utilizing photodiode induction to follow it corresponding to bandpass filter matched, thus achieve the optical component that be instead of some scattering devices in traditional light splitting technology by the multispectral sensor that structure is tiny, deliver more information exactly and reduce the volume of optical device, further, the integrated multiple bandpass filter in the surface due to multispectral sensor, and when corresponding optical wavelength selected by bandpass filter is different, just can realize detecting the intensity of the fluorescence signal of the multi-wavelength selected via bandpass filter simultaneously.In addition, miniaturized multichannel real-time fluorescence spectrum detection device of the present invention adopts the fluorescence transmission channel be made up of optical fiber or fibre bundle, every root optical fiber in optical fiber or fibre bundle forms an independently emitting fluorescence passage, reduce the energy loss of emitting fluorescence in paths, and the transmission mode of this fluorescence signal is cleaner than traditional open transmission structure, be not easy to by contamination by dust, improve accuracy and the sensitivity of experiment.
Accompanying drawing explanation
Fig. 1 is the Cleaning Principle schematic diagram of single channel quantitative real time PCR Instrument in prior art.
Fig. 2 is the structural representation of the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the preferred embodiment of the present invention 1 provides.
Fig. 3 is the structural representation one of the multispectral sensor in the subminiaturization hyperchannel real-time fluorescence spectrum detection device that provides of the preferred embodiment of the present invention 1.
Fig. 4 is the structural representation two of the multispectral sensor in the subminiaturization hyperchannel real-time fluorescence spectrum detection device that provides of the preferred embodiment of the present invention 1.
Fig. 5 is the structural representation of the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the preferred embodiment of the present invention 2 provides.
Fig. 6 is the structural representation of the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the preferred embodiment of the present invention 3 provides.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Fig. 2,5, the structural representation of the 6 subminiaturization hyperchannel real-time fluorescence spectrum detection devices provided for the present invention's three preferred embodiments.This subminiaturization hyperchannel real-time fluorescence spectrum detection device is applied to the test experience of laboratory or medical treatment, can be used in quantitatively detecting the enforcement of such as nucleic acid amplification reaction, but not as limit, in actual applications, described subminiaturization hyperchannel real-time fluorescence spectrum detection device also can be applicable to other real-time quantitatives based on fluorescence signal detection realization or qualitative detection experiment.
As shown in Figure 2, the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the preferred embodiment of the present invention provides comprises sample cell 10, excitation source 20, fluorescence transmission channel 30 and multispectral sensor 40.Wherein, hold in sample cell 10 and need test sample product and fluorescent material concentration dependent with testing sample.Excitation source 20 for the fluorescent material in excited sample pond, fluorescent material be stimulated light source produce excitation after produce utilizing emitted light (fluorescence).The fluorescence signal that fluorescence transmission channel 30 is formed for collecting utilizing emitted light, this fluorescence transmission channel 30 comprises the fibre bundle 31 be made up of optical fiber 32, and fluorescence signal is effectively exported by fibre bundle 31 and detects.Multispectral sensor 40 comprises multiple bandpass filter 41, and the photodiode 42 corresponding with bandpass filter quantity.Multispectral sensor 40 receives and detects the intensity transmitting the fluorescence signal come from fibre bundle 31, the integrated multiple bandpass filter in surface due to multispectral sensor 40, and when the corresponding optical wavelength selected by different bandpass filters is set to difference, just can realize the optical electivity to different wave length fluorescence, photodiode induces the fluorescence intensity of the wavelength corresponding to bandpass filter matched with it simultaneously.
During specific implementation, in sample cell 10, hold needing test sample product and fluorescent material concentration dependent with testing sample.The enforcement being applied to nucleic acid amplification reaction when subminiaturization hyperchannel real-time fluorescence spectrum detection device of the present invention quantitatively detects, testing sample is the object nucleic acid sheet degree produced in nucleic acid amplification reaction, and the fluorescent material relevant to object nucleic acid concentration is integrated in the Auele Specific Primer of nucleic acid amplification reaction usually, and in order to be that to be unlikely to not carrying out amplified reaction be that the light source activation that is namely excited produces fluorescence to the fluorescent material being connected to primer, usually the fluorescent quenching group in addition of nucleic acid amplification reaction primer is jointly connected to fluorescent material or fluorophor, thus the quantity of allow to be excited optical excitation fluorescent material or fluorophor is directly proportional to the concentration of object nucleic acid sheet degree in amplified reaction effectively, thus enable subminiaturization hyperchannel real-time fluorescence spectrum detection device of the present invention by detecting the intensity determination testing sample concentration of the fluorescence that is excited.Certainly, auxiliary with standard items in nucleic acid amplification reaction in order to determine that testing sample concentration also needs, determine testing sample concentration by analyzing and processing.Also can by effectively analyzing judgement to the test of fluorescence intensity signals to the qualitative features of testing sample and the validity of nucleic acid amplification reaction lacking standard items.Certainly, above-mentioned explanation is only quantitatively be detected as example with the enforcement of nucleic acid amplification reaction, sample cell of the present invention and corresponding reaction principle thereof are described, but not as limit, in actual applications, subminiaturization hyperchannel real-time fluorescence spectrum detection device of the present invention also can be applicable to other real-time quantitatives based on fluorescence signal detection realization or qualitative detection experiment.
As shown in Figure 2, in the embodiment of the present invention 1, fluorescence transmission channel comprises the mirror array 33 and convergent lens lens arra 34 that are arranged at described sample cell 10 both sides, multiple reflective mirror 331 be arranged in parallel is comprised in mirror array 33, convergent lenses array 34 comprises the convergent lens 341 corresponding with reflective mirror quantity, and each reflective mirror 331 and the convergent lens 341 matched form a phosphor collection unit.Fluorescence signal is sent to multispectral sensor 40 via fibre bundle 31 after being collected by this phosphor collection unit again.Analyze from light path, fluorescent material be stimulated light source produce utilizing emitted light (fluorescence) with diffusion mode produce, part direct projection is to convergent lens 341, and another part assembles the light entrance end to fibre bundle via convergent lens 341 after being reflected by reflective mirror 331 again.In order to make fluorescence effectively assemble, the reflective surface of the reflective mirror 331 adopted in this enforcement is arranged to the spill matched with the scope of its light collecting zone.
In the embodiment of the present invention 1, the optical fiber 32 that many light entrance ends are arranged as one is included in the fibre bundle 31 arranged in fluorescence transmission channel, the light entrance end of each optical fiber 32 is corresponding with a phosphor collection unit, and the every root optical fiber in fibre bundle all forms the fluorescence channel that an individual wavelengths is selected.As shown in Figures 3 and 4, multispectral sensor 40 comprises the multiple bandpass filters 41 being integrated in its surface, and the photodiode 42 corresponding with bandpass filter 41 quantity, and bandpass filter 41 is fitted the surface of photodiode 42; Each bandpass filter 41 and the photodiode 42 matched form a fluorescence intensity detecting unit, for detecting the fluorescence intensity of the wavelength corresponding to bandpass filter that this fluorescence intensity detecting unit is arranged, the light exit end of each optical fiber is corresponding with a fluorescence intensity detecting unit.In conjunction with in the present embodiment for the description of fluorescence transmission channel, the light entrance end of each optical fiber 32 is corresponding with a phosphor collection unit, every root optical fiber in fibre bundle all forms the fluorescence channel that an individual wavelengths is selected, the light exit end of each optical fiber is corresponding with a fluorescence intensity detecting unit simultaneously, therefore, comprise the phosphor collection unit that reflective mirror 331 forms with the convergent lens 341 matched, the fluorescence channel of optical fiber 32 correspondence and form fluorescence intensity detecting unit just one_to_one corresponding by bandpass filter 41 and the photodiode 42 that matches, thus realize the independent detection of the subchannel individual transmission of fluorescence signal and the different wave length fluorescence intensity selected by bandpass filter.
As in Figure 2-4, at the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the embodiment of the present invention 1 provides, its multispectral sensor 40 comprises the bandpass filter 41 that 8 are integrated in its surface, correspondingly, photodiode 42 is also provided with 8, bandpass filter 41 is fitted the surface of photodiode 42, each photodiode 42 and fit in bandpass filter 41 above it and form a fluorescence intensity detecting unit.Each fluorescence intensity detecting unit all can select independently emitting fluorescence wavelength, and detects the fluorescence intensity of the wavelength corresponding to bandpass filter that this fluorescence intensity detecting unit is arranged.And it should be noted that, in the solution of the present invention, effectively can realize that 8 even more bandpass filters (and photodiode corresponding with it) are integrated in area and be less than 1cm
2multispectral sensor, deliver more information exactly and and greatly reduce the volume of optical device.Correspondingly, 8 individual fibers 32 are provided with in 31 in fibre bundle, thus forming the fluorescence channel of 8 individual wavelengths selections, the light entrance end of each optical fiber 32 is corresponding with a phosphor collection unit, and the light exit end of each optical fiber is corresponding with a fluorescence intensity detecting unit.During specific implementation, by the light entrance end to optical fiber in fibre bundle 32, bandpass filter 41 in light exit end and multispectral sensor 40 is numbered one to one, each numbering correspondence one independently phosphor collection, transmit and sense channel, and each passage be all be connected by correspondence one by phosphor collection unit (convergent lens 341 comprising reflective mirror 331 and match), fluorescence channel optical fiber 32 of a transmission fluorescence (in the fibre bundle 31) and fluorescence intensity detecting unit (by photodiode 42 with fit in bandpass filter 41 above it and form) form, when containing multiple fluorescent substance in sample cell, and when multiple fluorescent substance is stimulated and produces the fluorescence of multiple different wave length, when the selection wavelength of bandpass filter 41 used in each passage is set to the consistent wavelength with a kind of fluorescence in multiple different wave length fluorescence, just can realize carrying out science to the experimental data of each passage by subminiaturization hyperchannel real-time fluorescence spectrum detection device of the present invention, analyze accurately, namely the situation of reaction multiple in sample cell is detected simultaneously.Certainly, in the embodiment of the present invention, the quantity of the bandpass filter quantity on the number of fibers in number of channels, fibre bundle, multispectral sensor surface, convergent lens quantity and reflective mirror is not limited to the quantity of corresponding construction used in technical scheme in other embodiments of the invention, when specific implementation, adjust the common technology means that the quantity of corresponding construction and arrangement mode also should be understood to this area as required, belong to the scope that the claims in the present invention contain.
In addition, for the setting of excitation source 20, be provided with an excitation source 20 in the embodiment of the present invention 1, in order to make the fluorescent material in sample cell effectively excite, the position of excitation source 20 and the irradiating angle of light source irradiate for foundation adjusts so that testing sample can be made more effectively to receive.In addition, in conjunction with physioptial ultimate principle, the frequency of excitation source is greater than utilizing emitted light (fluorescence) usually.When containing multiple fluorescent substance in sample cell, and when its fluorescence frequency being stimulated produced is different, the exciting light that this excitation source produces needs to guarantee that various fluorescent material all can effectively be excited and produces corresponding utilizing emitted light.And from the selection of light source, in the embodiment of the present invention, excitation source selects laser diode, the exciting light that excitation source can be made to send due to laser controls in single or very little frequency range, ensure to possess fluorescent material excite the intensity of the exciting light of ability and avoid light disturbance, thus improve accuracy and the sensitivity of experiment.Certainly in other embodiments of the invention also can as required with specific experiment condition, select common LED at interior other light sources as excitation source, but radiative light wave must be produced containing fluorescent material can be made effectively to excite in this excitation source.
About the quantity of excitation source, in the embodiment of the present invention 1, be provided with an excitation source, but in other embodiments of the invention, also can adjust the number of excitation source and position as required.As shown in Figure 5, in the embodiment of the present invention 2, excitation source 20 is provided with two, lay respectively at the both sides, both sides of sample cell, and the excitation wavelength difference that two excitation sources 20 produce, when containing multiple fluorescent substance in sample cell, different exciting lights can excite and produce different fluorescence signals, excitation source 20 is selected and is arranged just more targeted, and the frequency range of exciting light and power are according to controllability.Certainly, in other embodiments of the invention, also can as required excitation source be set to multiple, be in different objects, select the excitation source of different frequency that test reaction can be made according to specific aim and accuracy, sample cell testing sample and fluorescent material also can be made to be subject to more uniform exciting irradiation simultaneously.
As shown in Figure 6, the subminiaturization hyperchannel real-time fluorescence spectrum detection device that the embodiment of the present invention 3 provides, be set to many simple optical fibers 32 in its fluorescence transmission channel, the light entrance end of each simple optical fiber 32 is corresponding with phosphor collection unit and form the fluorescence channel that an individual wavelengths selects.Simultaneously, in the embodiment of the present invention 3, the fluorescent material held in each sample cell 10 is also set to one, excitation source 20 and sample cell 10 also one_to_one corresponding, each sample cell 10 correspondence is provided with a phosphor collection unit, the convergent lens 341 comprising reflective mirror (not shown) He match.During specific implementation, 8 simple optical fibers 32 be connected from different sample cell 10 phase are provided with in fluorescence transmission channel, corresponding with it, photodiode 42 is also provided with 8, bandpass filter 41 is fitted the surface of photodiode 42, each photodiode 42 and fit in bandpass filter 41 above it and form a fluorescence intensity detecting unit.Each fluorescence intensity detecting unit all can select independently emitting fluorescence wavelength, and detects the fluorescence intensity of the wavelength corresponding to bandpass filter that this fluorescence intensity detecting unit is arranged.Correspondingly, article 8, simple optical fiber 32 forms the fluorescence channel that 8 individual wavelengths are selected, the light entrance end of each simple optical fiber is corresponding with a phosphor collection unit, a sample cell and an excitation source, and the light exit end of each simple optical fiber 32 is corresponding with a fluorescence intensity detecting unit.By to sample cell 10, the light entrance end of optical fiber 32 in fibre bundle, bandpass filter 41 in light exit end and multispectral sensor 40 is numbered one to one, each numbering correspondence one independently phosphor collection, transmit and sense channel, and each passage be all be connected by correspondence one by phosphor collection unit (convergent lens 341 comprising reflective mirror 331 and match), a fluorescence channel (simple optical fiber 32) and fluorescence intensity detecting unit (by photodiode 42 and fit in bandpass filter 41 above it and form) form, so just can realize carrying out science to the experimental data of each passage by subminiaturization hyperchannel real-time fluorescence spectrum detection device of the present invention, analyze accurately.Meanwhile, containing a kind of fluorescent material in each sample cell 10, and the excitation source corresponding with this fluorescent material is set, thus realizes detecting single reaction in multiple different sample cell simultaneously.Certainly, in the embodiment of the present invention, the quantity of the bandpass filter quantity on number of channels, simple optical fiber quantity, multispectral sensor surface, convergent lens quantity and reflective mirror is not limited to the quantity of corresponding construction used in technical scheme in other embodiments of the invention, when specific implementation, adjust the common technology means that the quantity of corresponding construction and arrangement mode also should be understood to this area as required, belong to the scope that the claims in the present invention contain.
It should be noted that, the embodiment of the present invention additionally provides a kind of pick-up unit for laboratory or medical treatment, and it comprises the subminiaturization hyperchannel real-time fluorescence spectrum detection device disclosed in above-described embodiment.In addition, the subminiaturization hyperchannel real-time fluorescence spectrum detection device disclosed in this above-described embodiment is particularly useful for nucleic acid amplification test experience, therefore, embodiments of the invention additionally provide a kind of nucleic acid amplification detector, comprise the subminiaturization hyperchannel real-time fluorescence spectrum detection device disclosed in above-described embodiment.And control to be important technology index in nucleic acid amplification reaction due to temperature, such as regular-PCR temperature cycles formula amplification, and the climate control issues of isothermal amplification reactions is all extremely important, therefore is also provided with at described sample cell and realizes the temperature controlled temperature control probe of nucleic acid amplification reaction.
Compared to prior art, the subminiaturization hyperchannel real-time fluorescence spectrum detection device that above-described embodiment discloses, uses multispectral sensor to instead of traditional optical filter beam splitting system, greatly simplify the complexity of structure and reduce the volume of detection system.Multispectral sensor comprises bandpass filter and photodiode in the inventive solutions, and bandpass filter forms a fluorescence intensity detecting unit with the photodiode matched, utilize the bandpass filter being integrated in multispectral sensor surface to realize the optical electivity of different wave length, the fluorescence intensity of the wavelength utilizing photodiode induction to follow it corresponding to bandpass filter matched.Thus achieve the optical component that be instead of some scattering devices in traditional light splitting technology by the multispectral sensor that structure is tiny, deliver more information and reduce the volume of optical device; Further, the integrated multiple bandpass filter in the surface due to multispectral sensor, and when corresponding optical wavelength selected by bandpass filter is different, just can realize detecting the intensity of the fluorescence signal of the multi-wavelength selected via bandpass filter simultaneously.In addition, miniaturized multichannel real-time fluorescence spectrum detection device of the present invention adopts the fluorescence transmission channel be made up of optical fiber or fibre bundle, every root optical fiber in optical fiber or fibre bundle forms an independently emitting fluorescence passage, reduce the energy loss of emitting fluorescence in paths, and the transmission mode of this fluorescence signal is cleaner than traditional open transmission structure, be not easy to by contamination by dust, improve accuracy and the sensitivity of experiment.
The above is the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention.
Claims (10)
1. a subminiaturization hyperchannel real-time fluorescence spectrum detection device, is characterized in that, comprising:
Sample cell, holds needing test sample product and fluorescent material concentration dependent with testing sample in this sample cell;
Excitation source, for the fluorescent material in excited sample pond, fluorescent material be stimulated light source produce excitation after produce utilizing emitted light;
Fluorescence transmission channel, for collecting the fluorescence signal that utilizing emitted light is formed, the fibre bundle that this fluorescence transmission channel comprises optical fiber or is made up of optical fiber, fluorescence signal is effectively exported by optical fiber or fibre bundle and detects;
Multispectral sensor, described multispectral sensor comprises one or more bandpass filter and the photodiode corresponding with described bandpass filter quantity; This multispectral sensor receives and detects the intensity of the fluorescence signal come from the transmission of fluorescence transmission channel, described bandpass filter realizes the optical electivity of different wave length fluorescence, and photodiode induces the fluorescence intensity of the wavelength corresponding to bandpass filter matched with it.
2. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 1, is characterized in that: the fibre bundle that described fluorescence transmission channel comprises reflective mirror, convergent lens and optical fiber or is made up of optical fiber; Fluorescent material be stimulated light source produce utilizing emitted light direct projection or by after mirror reflection by convergent lens assemble to optical fiber or fibre bundle light entrance end.
3. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 1, it is characterized in that: described fluorescence transmission channel also comprises the mirror array and convergent lens lens arra that are arranged at described sample cell both sides, multiple reflective mirror be arranged in parallel is comprised in mirror array, the reflective surface of this reflective mirror is arranged to the spill matched with the scope of its light collecting zone, described convergent lenses array comprises the convergent lens corresponding with reflective mirror quantity, each reflective mirror and the convergent lens matched form a phosphor collection unit, fluorescence signal is sent to described multispectral sensor via optical fiber or the fibre bundle that is made up of optical fiber after being collected by this phosphor collection unit again.
4. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 3, it is characterized in that: in described fluorescence transmission channel, be provided with fibre bundle, the optical fiber that many light entrance ends are arranged as one is included in described fibre bundle, the light entrance end of each optical fiber is corresponding with a phosphor collection unit, and the every root optical fiber in fibre bundle all forms the fluorescence channel that an individual wavelengths is selected.
5. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 3, it is characterized in that: be provided with many simple optical fibers in described fluorescence transmission channel, the light entrance end of each simple optical fiber is corresponding with phosphor collection unit and form the fluorescence channel that an individual wavelengths selects.
6. the subminiaturization hyperchannel real-time fluorescence spectrum detection device as described in claim 4 or 5, it is characterized in that: described multispectral sensor comprises the multiple bandpass filters being integrated in its surface, and the photodiode corresponding with described bandpass filter quantity, described bandpass filter is fitted the surface of described photodiode; Each bandpass filter and the photodiode matched form a fluorescence intensity detecting unit, for detecting the fluorescence intensity of the wavelength corresponding to bandpass filter that this fluorescence intensity detecting unit is arranged; The light exit end of each optical fiber is corresponding with a fluorescence intensity detecting unit.
7. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 1, is characterized in that: more than described excitation source is provided with one or two, and plural excitation source produces the exciting light of different frequency.
8. subminiaturization hyperchannel real-time fluorescence spectrum detection device as claimed in claim 1, is characterized in that: described excitation source is for being laser diode or LED light source.
9., for a pick-up unit for laboratory or medical treatment, it is characterized in that: comprise subminiaturization hyperchannel real-time fluorescence spectrum detection device according to claim 1.
10. a nucleic acid amplification detector, is characterized in that: comprise subminiaturization hyperchannel real-time fluorescence spectrum detection device according to claim 1; Described sample cell is also provided with and realizes the temperature controlled temperature control probe of nucleic acid amplification reaction.
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| CN201510063198.2A CN104677870A (en) | 2015-02-06 | 2015-02-06 | Superminiaturization multi-channel real-time fluorescent spectrum detector |
| PCT/CN2016/072014 WO2016124083A1 (en) | 2015-02-06 | 2016-01-25 | Superminiature multi-channel real-time fluorescence spectrometer |
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