CN117402721B - Detection device and detection method for multicolor fluorescence detection - Google Patents
Detection device and detection method for multicolor fluorescence detection Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 57
- 238000001917 fluorescence detection Methods 0.000 title claims abstract description 57
- 230000003595 spectral effect Effects 0.000 claims abstract description 40
- 239000000975 dye Substances 0.000 claims description 95
- 230000005284 excitation Effects 0.000 claims description 64
- 238000005375 photometry Methods 0.000 claims description 34
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- 238000010438 heat treatment Methods 0.000 claims description 9
- 230000003287 optical effect Effects 0.000 claims description 9
- 210000004907 gland Anatomy 0.000 claims description 8
- 230000002596 correlated effect Effects 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 239000013307 optical fiber Substances 0.000 description 11
- 238000003752 polymerase chain reaction Methods 0.000 description 9
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- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
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- 238000000137 annealing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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- B01L2300/0627—Sensor or part of a sensor is integrated
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- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6484—Optical fibres
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Abstract
The invention provides a detection device for multicolor fluorescence detection, which comprises a fluorescence detection module; the fluorescence detection module comprises a light emitting component, a light measuring component and a light outlet hole; the light-emitting component comprises a multicolor light-emitting element and a light-emitting multi-pass filter; the light measuring assembly comprises a light measuring multi-pass filter; a dichroic mirror is arranged between the luminous multi-pass filter and the photometric multi-pass filter in an inclined 45-degree angle; one spectral range of the multicolor luminous element corresponds to one passband of the luminous multi-pass filter; each pass band of the light-emitting multi-pass filter and each pass band of the light-measuring multi-pass filter are not overlapped with each other; one passband of the dichroic mirror corresponds to one passband of the light-measuring multi-pass filter, and each passband of the light-emitting multi-pass filter and each passband of the dichroic mirror do not overlap each other. The invention realizes multicolor fluorescence detection of a single fluorescence detection module through the multicolor luminous element, the luminous multi-pass filter, the dichroic mirror and the photometric multi-pass filter, so that the detection device is compact in arrangement and low in cost.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to a detection device and a detection method for multicolor fluorescence detection.
Background
Molecular diagnostics is an important branch of in vitro diagnostics. PCR (Polymerase Chain Reaction polymerase chain reaction) technology is one of the most widely used technologies in the molecular diagnostic technology center. The PCR technology is one method of synthesizing specific DNA segment in vitro enzymatically, and consists of high temperature denaturation, low temperature annealing, proper temperature extension and other reactions in 1 period, and the PCR process is performed circularly to amplify the target DNA fast.
Specific fluorescent probes are added into a PCR reaction system, and the fluorescence intensity of the reaction system is in direct proportion to the amplification times of the DNA fragments. The fluorescence intensity is collected in real time in each PCR cycle, so that the amplification intensity of the DNA fragment can be obtained, and the real-time quantification of the PCR detection process is realized. Multiple detected targets (samples) can be detected simultaneously by a multiple fluorescence method, so that the detection efficiency of the DNA fragments is improved.
The Chinese patent application CN114181823A discloses a PCR instrument with multiple temperature control modules for asynchronously and selectively matching channels and a detection method thereof, wherein a plurality of monochromatic fluorescence detection channels are adopted to be connected in parallel by a multicolor fluorescence detection module, an amplification pore plate is transmitted to the fluorescence detection module through optical fibers, and the fluorescence detection module is driven by a belt guide rail to do linear motion to realize the scanning of all optical fibers. The fluorescence detection module adopts a scheme of a dichroic mirror, the wavelength of an excitation filter is L1, the wavelength of the dichroic mirror is L2, the wavelength of an emission filter is L3, and L1 is less than L2 and less than L3. The dichroic mirror reflects light having a wavelength less than L2 and transmits light having a wavelength greater than L2. The light emitted by the light emitting element is filtered by the excitation filter, reflected by the dichroic mirror, and emitted out of the detection module. The detected light is transmitted through the dichroic mirror, filtered by the emission filter, irradiated on the photosensitive element and detected in light intensity. Only one optical fiber is arranged between each detection channel and the tested tube, and excitation light and emission light in the optical fiber are transmitted bidirectionally.
The Chinese patent No. 209065925U discloses a multichannel fluorescent quantitative PCR instrument, an amplification pore plate is 96 holes, fluorescent detection modules are in multichannel and circular arrangement, and all test pore holes are scanned by driving rotation of a motor. The excitation tube and the detection tube are two separate tubes, and an optical fiber is arranged between each tube and the tested tube, so that 2 optical fibers, one for unidirectional excitation light and one for unidirectional emission light are needed for detecting a monochromatic fluorescence channel of a test tube.
In the prior art, multiple groups of optical fibers are required to be arranged for multicolor fluorescence detection, excitation light and emission light are transmitted by utilizing the multiple groups of optical fibers, the optical fibers are high in transmission cost and have loss of light intensity, and multiple monochromatic detection fluxes of the multicolor fluorescence detection device in the prior art are connected in parallel, so that the volume is large.
Disclosure of Invention
The invention provides a detection device and a detection method for multicolor fluorescence detection, which are used for solving the technical problems of high optical fiber transmission cost, light intensity loss, parallel connection of a plurality of monochromatic detection fluxes and large volume of the conventional multicolor fluorescence detection device.
The technical scheme provided by the invention is as follows:
An object of the present invention is to provide a detection device for multicolor fluorescence detection, which at least comprises a temperature control module and a fluorescence detection module;
The temperature control module comprises a plurality of test tube seats in an array, and each test tube seat is used for placing one test tube; each test tube seat is provided with a detection hole, and a plurality of detection holes of a plurality of test tube seats are positioned on the same straight line;
The fluorescence detection module at least comprises a light emitting component, a light measuring component and a light outlet hole; the fluorescence detection module is arranged at one side of the temperature control module, and the fluorescence detection module is configured to: reciprocating along the linear direction of the detection holes so that the light emergent holes are aligned with the detection holes of each test tube seat in sequence;
the light-emitting component at least comprises a multicolor light-emitting element and a light-emitting multi-pass filter; the light measuring assembly at least comprises a light measuring multi-pass filter and a light measuring element;
A dichroic mirror is arranged between the luminous multi-pass filter and the photometric multi-pass filter in a 45-degree inclined manner;
the optical axes of the light emitting component and the light measuring component are vertical, the light emitting multi-pass filter and the light outlet hole are positioned on the same side of the dichroic mirror, and the optical axis of the light emitting component and the axis of the light outlet hole are positioned on the same straight line;
The number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the luminous multi-pass filters, and the number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the photometric multi-pass filters; the passband number of the photometric multi-pass filter is equal to that of the dichroic mirror;
Wherein, a spectrum range of the multicolor luminous element corresponds to a passband of the luminous multi-pass filter;
wherein each pass band of the luminescent multi-pass filter and each pass band of the photometric multi-pass filter do not overlap each other;
Wherein, all pass bands of the luminous multi-pass filter are not overlapped with each other, and all pass bands of the photometric multi-pass filter are not overlapped with each other;
Wherein one passband of the dichroic mirror corresponds to one passband of the photometric multi-pass filter, and each passband of the luminescent multi-pass filter and each passband of the dichroic mirror do not overlap each other;
Wherein all pass bands of the dichroic mirror do not overlap each other.
In a preferred embodiment, the detection device further comprises a bracket, wherein a guide rail, a driving wheel and a driven wheel are fixed on the bracket, and a belt is arranged between the driving wheel and the driven wheel in a surrounding manner;
The belt is fixed with the fluorescence detection module, and the fluorescence detection module is slidably arranged with the guide rail;
The driving wheel drives the belt to move, and the belt drives the fluorescent detection module to reciprocate along the straight line direction of the detection holes.
In a preferred embodiment, the temperature control module further comprises a heating plate, a temperature control element and a heat sink;
the heating plate is arranged on the temperature control element, the temperature control element is arranged on the radiator, and a plurality of test tube seats are arranged on the heating plate in an array mode.
In a preferred embodiment, the light emitting assembly further includes a light emitting lens disposed between the multicolor light emitting element and the light emitting multi-pass filter for transmitting excitation light of different spectral ranges emitted by the multicolor light emitting element to the light emitting multi-pass filter;
the light measuring assembly further comprises a light measuring lens, wherein the light measuring lens is arranged between the light measuring multi-pass filter and the light measuring element and is used for transmitting fluorescence with different wavelengths separated by the light measuring multi-pass filter to the light measuring element;
The fluorescent detection module further comprises a front gland, the front gland is provided with the light outlet hole, a light outlet lens is arranged in the front gland, and the light outlet lens is opposite to the light outlet hole.
In a preferred embodiment, the tube houses a plurality of different dyes and a target to be measured, the fluorescence detection intensity of each dye being positively correlated with the concentration of the target to be measured.
In a preferred embodiment, one passband of the luminescent multi-pass filter corresponds to the excitation wavelength of one of the dyes; one passband of the photometric multi-pass filter corresponds to the fluorescence wavelength of one of the dyes.
Another aspect of the present invention provides a detection method for multicolor fluorescence detection, the detection method comprising the following method steps:
the multicolor luminous element emits excitation light with different spectrum ranges;
the luminous multi-pass filter separates the excitation light of different spectrum ranges of the multicolor luminous element into excitation monochromatic light with different wavelengths;
The dichroic mirror vertically reflects the excited monochromatic light with different wavelengths to the light outlet hole, and the excited monochromatic light is injected into the test tube through the detection hole of the test tube seat to excite a plurality of different dyes in the test tube to generate fluorescence with different wavelengths;
Fluorescence of different wavelengths is reflected to the dichroic mirror, and the dichroic mirror transmits the fluorescence of different wavelengths to the photometry multi-pass filter;
The light measuring multi-pass filter separates fluorescence with different wavelengths and transmits the fluorescence with different wavelengths to the light measuring element;
The number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the luminous multi-pass filters, and the number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the photometric multi-pass filters; the passband number of the photometric multi-pass filter is equal to that of the dichroic mirror;
Wherein, a spectrum range of the multicolor luminous element corresponds to a passband of the luminous multi-pass filter;
wherein each pass band of the luminescent multi-pass filter and each pass band of the photometric multi-pass filter do not overlap each other;
Wherein, all pass bands of the luminous multi-pass filter are not overlapped with each other, and all pass bands of the photometric multi-pass filter are not overlapped with each other;
Wherein one passband of the dichroic mirror corresponds to one passband of the photometric multi-pass filter, and each passband of the luminescent multi-pass filter and each passband of the dichroic mirror do not overlap each other;
Wherein all pass bands of the dichroic mirror do not overlap each other.
In a preferred embodiment, a luminescent lens is disposed between the multicolor luminescent element and the luminescent multi-pass filter, and is used for transmitting the excitation light with different spectral ranges emitted by the multicolor luminescent element to the luminescent multi-pass filter;
and a light measuring lens is arranged between the light multi-pass filter and the light measuring element and is used for transmitting fluorescence with different wavelengths separated by the light measuring multi-pass filter to the light measuring element.
In a preferred embodiment, the tube houses a plurality of different dyes and a target to be measured, the fluorescence detection intensity of each dye being positively correlated with the concentration of the target to be measured.
In a preferred embodiment, one passband of the luminescent multi-pass filter corresponds to the excitation wavelength of one of the dyes; one passband of the photometric multi-pass filter corresponds to the fluorescence wavelength of one of the dyes.
Compared with the prior art, the technical scheme of the invention has at least the following beneficial effects:
The invention provides a detection device and a detection method for multicolor fluorescence detection, wherein multicolor fluorescence detection of a single fluorescence detection module is realized through a multicolor luminous element, a luminous multi-pass filter, a dichroic mirror and a photometric multi-pass filter, optical fiber transmission is omitted, scanning is performed through the single fluorescence detection module, and the device is compact in arrangement and low in cost.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram showing the overall structure of a detection device for multicolor fluorescence detection according to the present invention.
Fig. 2 is a schematic structural diagram of a temperature control module according to the present invention.
FIG. 3 is a schematic diagram of a fluorescence detection module according to the present invention.
FIG. 4 is a cross-sectional view of a temperature control module and fluorescence detection module arrangement of the present invention.
FIG. 5 is a cross-sectional view of a fluorescence detection module of the present invention.
FIG. 6 is a schematic diagram showing the relationship among the spectral range of a multicolor light-emitting element, the passband of a light-emitting multi-pass filter, the passband of a photometric multi-pass filter, and the passband of a dichroic mirror in one embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. It will be apparent that the described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by a person skilled in the art without creative efforts, based on the described embodiments of the present invention fall within the protection scope of the present invention.
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. Likewise, the terms "a," "an," or "the" and similar terms do not denote a limitation of quantity, but rather denote the presence of at least one. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect.
It should be noted that "upper", "lower", "left", "right", "front", "rear", and the like are used in the present invention only to indicate a relative positional relationship, and when the absolute position of the object to be described is changed, the relative positional relationship may be changed accordingly.
Referring to fig. 1 to 6, according to an embodiment of the present invention, there is provided a detection apparatus for multicolor fluorescence detection, including a bracket 1, a temperature control module 2, and a fluorescence detection module 3.
The support 1 is provided with a fixed guide rail 4, a driving wheel 6 and a driven wheel 7, and a belt 8 is arranged between the driving wheel 6 and the driven wheel 7 in a surrounding way. The belt 8 is fixed with the fluorescence detection module 3 through the belt pressing sheet 9, and the fluorescence detection module 3 is slidably mounted with the guide rail 4.
As shown in fig. 2, the temperature control module 2 includes a heating plate 201, a temperature control element 202, and a heat sink 203. The heating plate 201 is placed on the temperature control element 202, and the temperature control element 202 is placed on the heat sink 203.
The temperature control module 2 includes a plurality of test tube seats 2011 in an array. Specifically, a plurality of test tube holders 2011 are arrayed on the heating plate 201. Each tube holder 2011 is used for placing one test tube 10, each tube holder 2011 is provided with a test hole 2012, and the test holes 2012 of the tube holders 2011 are in the same straight line Z, as shown in fig. 1.
As shown in fig. 3, the fluorescence detection module 3 includes a front cover 301, a light-emitting housing 302, and a photometry housing 303. A light emitting component (described in detail below) is disposed in the light emitting housing 302, a light measuring component (described in detail below) is disposed in the light measuring housing 303, and a light emitting hole 3011 is formed in the front cover 301.
As shown in fig. 1, the fluorescence detection module 3 is arranged on the side of the temperature control module 2, and the fluorescence detection module 3 is configured to: the light-emitting holes 3011 are aligned with the test holes 2012 of each cuvette holder 2011 in sequence by reciprocating in the direction of the straight line (direction of the straight line Z) where the plurality of test holes 2012 are located (as indicated by arrow a in fig. 1).
Further, the bracket 1 fixes the stepper motor 5, the stepper motor 5 is connected with the driving wheel 6, the stepper motor 5 drives the driving wheel 6 to rotate, the driving wheel 6 drives the belt 8 to move, and the belt 8 drives the fluorescence detection module 3 to reciprocate along the linear direction (the direction of the straight line Z) where the plurality of detection holes 2012 are located, so that the light outlet holes 3011 are aligned with the detection holes 2012 of each test tube holder 2011 in sequence.
Referring to fig. 3, 4 and 5, according to an embodiment of the present invention, a light emitting assembly is disposed in a light emitting housing 302, the light emitting assembly including a multicolor light emitting element 304, a light emitting circuit board 305, a light emitting lens 306 and a light emitting multi-pass filter 307.
The light-emitting circuit board 305 is fixed to the light-emitting housing 302, the multicolor light-emitting element 304 is connected to the light-emitting circuit board 305, and the light-emitting lens 306 is interposed between the multicolor light-emitting element 304 and the light-emitting multi-pass filter 307. A luminescent lens 306 for transmitting the excitation light of different spectral ranges emitted by the multicolor luminescent element 304 to a luminescent multi-pass filter 307.
A photometry module is disposed in the photometry housing 303, and the photometry module includes a photometry multi-pass filter 311, a photometry lens 310, a photometry element 308, and a photometry circuit board 309.
The photometry circuit board 309 is fixed to the photometry housing 303, the photometry element 308 is connected to the photometry circuit board 309, and the photometry lens 310 is disposed between the photometry multi-pass filter 311 and the photometry element 308. The photometry lens 310 is used for transmitting fluorescence with different wavelengths separated by the photometry multi-pass filter 311 to the photometry element 308.
According to an embodiment of the present invention, a dichroic mirror 312 is disposed between the light emitting multi-pass filter 307 and the photometric multi-pass filter 311 at an angle of 45 degrees. The dichroic mirror 312 disposed at an angle of 45 degrees is equivalent to a 45-degree incident filter, and light within its passband can be directly transmitted, and light of the stopband is reflected and emitted in a direction perpendicular to the incident light.
The front gland 301 is provided with a light outlet 3011, a light outlet lens 313 is arranged in the front gland 301, and the light outlet 313 is opposite to the light outlet 3011.
As shown in fig. 5, according to the embodiment of the present invention, the optical axes of the light emitting element and the photometry element are perpendicular, and the light emitting multi-pass filter 307 is located on the same side of the dichroic mirror 312 as the light emitting hole 3011, and the optical axis of the light emitting element is in the same straight line as the axis of the light emitting hole 3011.
That is, the optical axes of the multicolor light-emitting element 304, the light-emitting lens 306, and the light-emitting multi-pass filter 307 are perpendicular to the optical axes of the photometry multi-pass filter 311, the photometry lens 310, and the photometry element 308, and the optical axes of the photometry multi-pass filter 311, the photometry lens 310, and the photometry element 308 coincide with the axes of the light-emitting lens 313, and the light-emitting hole 3011.
According to an embodiment of the present invention, the number of spectral ranges of the multicolor light-emitting element 304 is equal to the number of pass bands of the light-emitting multi-pass filter 307; the number of spectrum ranges of the multicolor luminous element 304 is equal to the number of pass bands of the photometry multi-pass filter 311; the number of pass bands of the photometric multi-pass filter 311 is equal to the number of pass bands of the dichroic mirror 312. The tube 10 houses a plurality of different dyes and a target to be measured, and the fluorescence detection intensity of each dye is positively correlated with the concentration of the target to be measured.
As shown in fig. 6, three different dyes and a target (sample) to be measured are placed in the test tube 10 illustrated in this embodiment, and the fluorescence detection intensity of each dye is positively correlated with the concentration of the target (sample) to be measured.
The multicolor light-emitting element 304 emits excitation light in three different spectral ranges, that is, the multicolor light-emitting element 304 emits excitation light in three different spectral ranges: excitation light of the first spectral range P1, excitation light of the second spectral range P2, and excitation light of the third spectral range P3.
The light-emitting multi-pass filter 307 has three pass bands, that is, the light-emitting multi-pass filter 307 has three pass bands of: the light-emitting multi-pass filter comprises a first passband F1, a second passband F2 and a third passband F3. The three pass bands of the luminescent multi-pass filter 307 separate the excitation light of three different spectral ranges into excitation monochromatic light of three wavelengths.
The photometric multi-pass filter 311 has three pass bands, that is, the photometric multi-pass filter 311 has three pass bands of: the light measuring multi-pass filter comprises a light measuring multi-pass filter first passband C1, a light measuring multi-pass filter second passband C2 and a light measuring multi-pass filter third passband C3. The three pass bands of the photometric multi-pass filter 311 separate the three fluorescent dyes in the cuvette 10 into three wavelengths of fluorescence.
The dichroic mirror 312 has three pass bands, i.e., the dichroic mirror 312 has three pass bands of: a dichroic mirror first passband D1, a dichroic mirror second passband D2, and a dichroic mirror third passband D3.
According to an embodiment of the present invention, one spectral range of the multicolor light-emitting element 304 corresponds to one passband of the light-emitting multi-pass filter 307.
In the present embodiment, the first spectral range P1 of the multicolor light emitting element 304 corresponds to the first pass band F1 of the light emitting multi-pass filter 307, and the first spectral range P1 of the multicolor light emitting element 304 does not overlap with the second pass band F2 of the light emitting multi-pass filter 307, and the first spectral range P1 of the multicolor light emitting element 304 does not overlap with the third pass band F3 of the light emitting multi-pass filter 307.
Similarly, the second spectral range P2 of the multicolor light emitting element 304 corresponds to the second pass band F2 of the light emitting multi-pass filter 307, and the second spectral range P2 of the multicolor light emitting element 304 does not overlap the first pass band F1 of the light emitting multi-pass filter 307, and the second spectral range P2 of the multicolor light emitting element 304 does not overlap the third pass band F3 of the light emitting multi-pass filter 307.
Similarly, the third spectral range P3 of the multicolor light emitting element 304 corresponds to the third passband F3 of the light emitting multi-pass filter 307, and the third spectral range P3 of the multicolor light emitting element 304 does not overlap the first passband F1 of the light emitting multi-pass filter 307, and the third spectral range P3 of the multicolor light emitting element 304 does not overlap the second passband F3 of the light emitting multi-pass filter 307.
According to an embodiment of the present invention, each pass band of the light emitting multi-pass filter 307 and each pass band of the photometry multi-pass filter 311 do not overlap each other.
In the present embodiment, the light-emitting multi-pass filter first passband F1 of the light-emitting multi-pass filter 307 is not overlapped with the light-measuring multi-pass filter first passband C1 of the light-measuring multi-pass filter 311; the light-emitting multi-pass filter first passband F1 of the light-emitting multi-pass filter 307 and the light-measuring multi-pass filter second passband C2 of the light-measuring multi-pass filter 311 do not overlap; the light-emitting multi-pass filter first pass band F1 of the light-emitting multi-pass filter 307 and the light-measuring multi-pass filter third pass band C3 of the light-measuring multi-pass filter 311 do not overlap.
The light-emitting multi-pass filter second passband F2 of the light-emitting multi-pass filter 307 does not overlap the light-measuring multi-pass filter first passband C1 of the light-measuring multi-pass filter 311; the light-emitting multi-pass filter second passband F2 of the light-emitting multi-pass filter 307 does not overlap with the photometry multi-pass filter second passband C2 of the photometry multi-pass filter 311; the light-emitting multi-pass filter second pass band F2 of the light-emitting multi-pass filter 307 does not overlap the light-measuring multi-pass filter third pass band C3 of the light-measuring multi-pass filter 311.
The third pass band F3 of the light-emitting multi-pass filter 307 does not overlap the first pass band C1 of the photometry multi-pass filter 311; the third passband F3 of the luminescent multi-pass filter 307 is not overlapped with the second passband C2 of the photometric multi-pass filter 311; the light-emitting multi-pass filter third passband F3 of the light-emitting multi-pass filter 307 does not overlap the light-measuring multi-pass filter third passband C3 of the light-measuring multi-pass filter 311.
According to the embodiment of the present invention, all pass bands of the light emitting multi-pass filter 307 are not overlapped with each other, and all pass bands of the photometric multi-pass filter 311 are not overlapped with each other.
In the present embodiment, the first pass band F1 of the light-emitting multi-pass filter, the second pass band F2 of the light-emitting multi-pass filter, and the third pass band F3 of the light-emitting multi-pass filter are not overlapped with each other, and the first pass band C1 of the light-measuring multi-pass filter, the second pass band C2 of the light-measuring multi-pass filter, and the third pass band C3 of the light-measuring multi-pass filter 311 are not overlapped with each other.
According to an embodiment of the present invention, one passband of the dichroic mirror 312 corresponds to one passband of the light-measuring multi-pass filter 311, and each passband of the light-emitting multi-pass filter 307 and each passband of the dichroic mirror 312 do not overlap each other.
In the present embodiment, the dichroic mirror first passband D1 of the dichroic mirror 312 corresponds to the photometric multi-pass filter first passband C1 of the photometric multi-pass filter 311, that is, the dichroic mirror first passband D1 of the dichroic mirror 312 includes the photometric multi-pass filter first passband C1 of the photometric multi-pass filter 311.
The dichroic mirror second passband D2 of the dichroic mirror 312 corresponds to the photometric multi-pass filter second passband C2 of the photometric multi-pass filter 311, i.e. the dichroic mirror second passband D2 of the dichroic mirror 312 comprises the photometric multi-pass filter second passband C2 of the photometric multi-pass filter 311.
The third dichroic mirror passband D3 of the dichroic mirror 312 corresponds to the third light-measuring bandpass filter passband C3 of the light-measuring bandpass filter 311, i.e., the third dichroic mirror passband D3 of the dichroic mirror 312 includes the third light-measuring bandpass filter passband C3 of the light-measuring bandpass filter 311.
The light emission multi-pass filter first pass band F1 of the light emission multi-pass filter 307 does not overlap with the dichroic mirror first pass band D1 of the dichroic mirror 312; the light-emitting multi-pass filter first pass band F1 of the light-emitting multi-pass filter 307 and the dichroic mirror second pass band D2 of the dichroic mirror 312 do not overlap; the light emission multi-pass filter first pass band F1 of the light emission multi-pass filter 307 does not overlap the dichroic mirror third pass band D3 of the dichroic mirror 312.
The light emission multi-pass filter second passband F2 of the light emission multi-pass filter 307 does not overlap the dichroic mirror first passband D1 of the dichroic mirror 312; the light-emitting multi-pass filter second pass band F2 of the light-emitting multi-pass filter 307 does not overlap the dichroic mirror second pass band D2 of the dichroic mirror 312; the light emission multi-pass filter second pass band F2 of the light emission multi-pass filter 307 does not overlap the dichroic mirror third pass band D3 of the dichroic mirror 312.
The light-emitting multi-pass filter third passband F3 of the light-emitting multi-pass filter 307 does not overlap the dichroic mirror first passband D1 of the dichroic mirror 312; the light-emitting multi-pass filter third passband F3 of the light-emitting multi-pass filter 307 does not overlap the dichroic mirror second passband D2 of the dichroic mirror 312; the light emission multi-pass filter third passband F3 of the light emission multi-pass filter 307 does not overlap the dichroic mirror third passband D3 of the dichroic mirror 312.
According to an embodiment of the present invention, all pass bands of dichroic mirror 312 do not overlap each other.
In the present embodiment, the dichroic mirror first passband D1 of the dichroic mirror 312, the dichroic mirror second passband D2 of the dichroic mirror 312, and the dichroic mirror third passband D3 of the dichroic mirror 312 do not overlap with each other.
According to an embodiment of the present invention, one passband of the luminescent multi-pass filter 307 corresponds to the excitation wavelength of one dye; one pass band of the photometric multi-pass filter 311 corresponds to the fluorescence wavelength of one dye.
In this embodiment, the three different dyes are a first dye, a second dye, and a third dye, respectively. The first dye of the first dye has an excitation wavelength of J1, the second dye of the second dye has an excitation wavelength of J2, and the third dye of the third dye has an excitation wavelength of J3.
The first dye of the first dye has a fluorescence wavelength of Y1, the second dye of the second dye has a fluorescence wavelength of Y2, and the third dye of the third dye has a fluorescence wavelength of Y3.
The first pass band F1 of the luminescent multi-pass filter 307 corresponds to the first dye excitation wavelength J1 of the first dye, i.e., the first pass band F1 of the luminescent multi-pass filter 307 comprises the first dye excitation wavelength J1 of the first dye.
The second pass band F2 of the luminescent multi-pass filter 307 corresponds to the second dye excitation wavelength J2 of the second dye, i.e., the second pass band F2 of the luminescent multi-pass filter 307 comprises the second dye excitation wavelength J2 of the second dye.
The third pass band F3 of the light-emitting multi-pass filter 307 corresponds to the third dye excitation wavelength J3 of the third dye, i.e., the third pass band F3 of the light-emitting multi-pass filter 307 comprises the third dye excitation wavelength J3 of the third dye.
The first pass band C1 of the photometric multi-pass filter 311 corresponds to the fluorescent wavelength Y1 of the first dye, i.e. the first pass band C1 of the photometric multi-pass filter 311 contains the fluorescent wavelength Y1 of the first dye.
The second pass band C2 of the photometric multi-pass filter 311 corresponds to the fluorescent wavelength Y2 of the second dye, i.e. the second pass band C21 of the photometric multi-pass filter 311 comprises the fluorescent wavelength Y2 of the second dye.
The third passband C3 of the photometric multi-pass filter 311 corresponds to the third dye fluorescence wavelength Y3 of the third dye, i.e. the third passband C3 of the photometric multi-pass filter 311 contains the third dye fluorescence wavelength Y3 of the third dye.
According to an embodiment of the present invention, there is provided a detection method of multicolor fluorescence detection, the detection method including the method steps of:
The multicolor light emitting element 304 emits excitation light of different spectral ranges.
According to the embodiment of the present invention, the number of spectral ranges of the multicolor light emitting element 304 is equal to the number of pass bands of the light emitting multi-pass filter 307, and the number of spectral ranges of the multicolor light emitting element 304 is equal to the number of pass bands of the light measuring multi-pass filter 311; the number of pass bands of the photometric multi-pass filter 311 is equal to the number of pass bands of the dichroic mirror 312.
One spectral range of the multicolor light-emitting element 304 corresponds to one passband of the light-emitting multi-pass filter 307. Each pass band of the light emission multi-pass filter 307 and each pass band of the photometry multi-pass filter 311 do not overlap each other.
One passband of the dichroic mirror 312 corresponds to one passband of the light-measuring multi-pass filter 311, and each passband of the light-emitting multi-pass filter 307 and each passband of the dichroic mirror 312 do not overlap each other;
All pass bands of the light emission multi-pass filter 307 do not overlap each other, all pass bands of the photometry multi-pass filter 311 do not overlap each other, and all pass bands of the dichroic mirror 312 do not overlap each other.
One passband of the luminescent multi-pass filter 307 corresponds to the excitation wavelength of one dye; one pass band of the photometric multi-pass filter 311 corresponds to the fluorescence wavelength of one dye.
In this embodiment, three different dyes and targets (samples) to be tested are placed in the test tube 10, and the multicolor light emitting element 304 emits excitation light in three different spectral ranges.
The light-emitting circuit board 305 controls excitation light of three spectral ranges (a first spectral range P1, a second spectral range P2, and a third spectral range P3) emitted from the multicolor light-emitting element 402, and transmits the excitation light to the light-emitting multipass filter 307 via the light-emitting lens 306.
The luminescent multi-pass filter 307 separates excitation light of different spectral ranges from the multicolor luminescent element 304 into excitation monochromatic light of different wavelengths.
In the present embodiment, the light-emitting lens 306 transmits the excitation light of the three spectral ranges (the first spectral range P1, the second spectral range P2, and the third spectral range P3) emitted by the multicolor light-emitting element 304 to the light-emitting multi-pass filter 307.
Since one passband of the light-emitting multi-pass filter 307 corresponds to the excitation wavelength of one dye, the light-emitting multi-pass filter first passband F1 of the light-emitting multi-pass filter 307 separates the excitation light of the first spectral range P1 of the multicolor light-emitting element 304 into excitation monochromatic light of the first dye excitation wavelength J1;
The second pass band F2 of the luminescent multi-pass filter 307 separates the excitation light of the second spectrum range P2 of the multicolor luminescent element 304 into excitation monochromatic light of the second dye excitation wavelength J2;
The third pass band F3 of the luminescent multi-pass filter 307 separates the excitation light of the third spectral range P3 of the multicolor light emitting element 304 into excitation monochromatic light of the third dye excitation wavelength J3.
The dichroic mirror 312 vertically reflects the excited monochromatic light with different wavelengths to the light emitting hole 3011, and emits the excited monochromatic light into the cuvette 10 through the detection hole 2012 of the cuvette holder 2011, so as to excite multiple different dyes in the cuvette 10 to generate fluorescence with different wavelengths.
Since each pass band of the light-emitting multi-pass filter 307 and each pass band of the dichroic mirror 312 do not overlap with each other, in this embodiment, the dichroic mirror 312 vertically reflects the excitation monochromatic light of three wavelengths (the excitation monochromatic light of the first dye excitation wavelength J1, the excitation monochromatic light of the second dye excitation wavelength J2, and the excitation monochromatic light of the third dye excitation wavelength J3) to the light-emitting lens 313, and emits the light into the cuvette 10 through the light-emitting hole 3011 and the detection hole 2012 of the cuvette holder 2011, so that the three different dyes in the excitation cuvette 10 generate fluorescence of three different wavelengths.
That is, the three dyes respectively generate a first dye having a fluorescence wavelength of Y1, a second dye having a fluorescence wavelength of Y2, and a third dye having a fluorescence wavelength of Y3.
The fluorescence of different wavelengths is reflected to the dichroic mirror 312, and the dichroic mirror 312 transmits the fluorescence of different wavelengths to the photometric multi-pass filter 311.
In this embodiment, three kinds of fluorescence with different wavelengths (first dye fluorescence wavelength Y1 of the first dye, second dye fluorescence wavelength Y2 of the second dye, and third dye fluorescence wavelength Y3 of the third dye) are reflected to the dichroic mirror 312 through the detection hole 2012, the light emitting hole 3011, and the light emitting lens 313 of the cuvette holder 2011. Since one pass band of the dichroic mirror 312 corresponds to one pass band of the photometric multi-pass filter 311, one pass band of the photometric multi-pass filter 311 corresponds to the fluorescence wavelength of one dye, and fluorescence of three different wavelengths (fluorescence of the first dye fluorescence wavelength Y1 of the first dye, fluorescence of the second dye fluorescence wavelength Y2 of the second dye, fluorescence of the third dye fluorescence wavelength Y3 of the third dye) is transmitted to the photometric multi-pass filter 311 through the dichroic mirror 312.
The photometric multi-pass filter 311 separates fluorescence of different wavelengths, and transmits the fluorescence of different wavelengths to the photometric element.
In this embodiment, the light-measuring multi-pass filter 311 separates the fluorescence of three wavelengths (the fluorescence of the first dye fluorescence wavelength Y1, the fluorescence of the second dye fluorescence wavelength Y2 and the fluorescence of the third dye fluorescence wavelength Y3), and transmits the fluorescence of three wavelengths (the fluorescence of the first dye fluorescence wavelength Y1, the fluorescence of the second dye fluorescence wavelength Y2 and the fluorescence of the third dye fluorescence wavelength Y3) to the light-measuring element 308 via the light-measuring lens 310, and the light-measuring element 308 converts the fluorescence of three wavelengths into an electrical signal and transmits the electrical signal to the light-measuring circuit board 309 for multicolor fluorescence detection.
The following points need to be described:
(1) The drawings of the embodiments of the present invention relate only to the structures related to the embodiments of the present invention, and other structures may refer to the general designs.
(2) In the drawings for describing embodiments of the present invention, the thickness of layers or regions is exaggerated or reduced for clarity, i.e., the drawings are not drawn to actual scale. It will be understood that when an element such as a layer, film, region or substrate is referred to as being "on" or "under" another element, it can be "directly on" or "under" the other element or intervening elements may be present.
(3) The embodiments of the invention and the features of the embodiments can be combined with each other to give new embodiments without conflict.
The present invention is not limited to the above embodiments, but the scope of the invention is defined by the claims.
Claims (6)
1. The detection device for multicolor fluorescence detection is characterized by at least comprising a temperature control module and a fluorescence detection module;
The temperature control module comprises a plurality of test tube seats in an array, and each test tube seat is used for placing one test tube; each test tube seat is provided with a detection hole, and a plurality of detection holes of a plurality of test tube seats are positioned on the same straight line;
The fluorescence detection module at least comprises a light emitting component, a light measuring component and a light outlet hole; the fluorescence detection module is arranged at one side of the temperature control module, and the fluorescence detection module is configured to: reciprocating along the linear direction of the detection holes so that the light emergent holes are aligned with the detection holes of each test tube seat in sequence;
the light-emitting component at least comprises a multicolor light-emitting element and a light-emitting multi-pass filter; the light measuring assembly at least comprises a light measuring multi-pass filter and a light measuring element;
A dichroic mirror is arranged between the luminous multi-pass filter and the photometric multi-pass filter in a 45-degree inclined manner;
the optical axes of the light emitting component and the light measuring component are vertical, the light emitting multi-pass filter and the light outlet hole are positioned on the same side of the dichroic mirror, and the optical axis of the light emitting component and the axis of the light outlet hole are positioned on the same straight line;
The number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the luminous multi-pass filters, and the number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the photometric multi-pass filters; the passband number of the photometric multi-pass filter is equal to that of the dichroic mirror;
Wherein, a spectrum range of the multicolor luminous element corresponds to a passband of the luminous multi-pass filter;
wherein each pass band of the luminescent multi-pass filter and each pass band of the photometric multi-pass filter do not overlap each other;
Wherein, all pass bands of the luminous multi-pass filter are not overlapped with each other, and all pass bands of the photometric multi-pass filter are not overlapped with each other;
wherein, the test tube is internally provided with a plurality of different dyes and a tested target, and the fluorescence detection intensity of each dye is positively correlated with the concentration of the tested target;
One passband of the luminescent multi-pass filter corresponds to the excitation wavelength of one of the dyes; one passband of the dichroic mirror corresponds to one passband of the photometric multi-pass filter, and one passband of the photometric multi-pass filter corresponds to a fluorescence wavelength of the dye; and each pass band of the light-emitting multi-pass filter and each pass band of the dichroic mirror do not overlap each other;
Wherein all pass bands of the dichroic mirror do not overlap each other.
2. The detection device according to claim 1, further comprising a bracket, wherein the bracket is provided with a fixed guide rail, a driving wheel and a driven wheel, and a belt is arranged between the driving wheel and the driven wheel in a surrounding manner;
The belt is fixed with the fluorescence detection module, and the fluorescence detection module is slidably arranged with the guide rail;
The driving wheel drives the belt to move, and the belt drives the fluorescent detection module to reciprocate along the straight line direction of the detection holes.
3. The detection device of claim 1, wherein the temperature control module further comprises a heating plate, a temperature control element, and a heat sink;
the heating plate is arranged on the temperature control element, the temperature control element is arranged on the radiator, and a plurality of test tube seats are arranged on the heating plate in an array mode.
4. The detection device according to claim 1, wherein the light emitting assembly further comprises a light emitting lens disposed between the multicolor light emitting element and the light emitting multi-pass filter for transmitting excitation light of different spectral ranges emitted by the multicolor light emitting element to the light emitting multi-pass filter;
the light measuring assembly further comprises a light measuring lens, wherein the light measuring lens is arranged between the light measuring multi-pass filter and the light measuring element and is used for transmitting fluorescence with different wavelengths separated by the light measuring multi-pass filter to the light measuring element;
The fluorescent detection module further comprises a front gland, the front gland is provided with the light outlet hole, a light outlet lens is arranged in the front gland, and the light outlet lens is opposite to the light outlet hole.
5. A detection method for multicolor fluorescence detection, characterized in that the detection method comprises the following method steps:
the multicolor luminous element emits excitation light with different spectrum ranges;
the luminous multi-pass filter separates the excitation light of different spectrum ranges of the multicolor luminous element into excitation monochromatic light with different wavelengths;
The dichroic mirror vertically reflects the excited monochromatic light with different wavelengths to the light outlet hole, and the excited monochromatic light is injected into the test tube through the detection hole of the test tube seat to excite a plurality of different dyes in the test tube to generate fluorescence with different wavelengths;
Fluorescence of different wavelengths is reflected to the dichroic mirror, and the dichroic mirror transmits the fluorescence of different wavelengths to the photometry multi-pass filter;
The light measuring multi-pass filter separates fluorescence with different wavelengths and transmits the fluorescence with different wavelengths to the light measuring element;
The number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the luminous multi-pass filters, and the number of the spectrum ranges of the multicolor luminous elements is equal to the number of the pass bands of the photometric multi-pass filters; the passband number of the photometric multi-pass filter is equal to that of the dichroic mirror;
Wherein, a spectrum range of the multicolor luminous element corresponds to a passband of the luminous multi-pass filter;
wherein each pass band of the luminescent multi-pass filter and each pass band of the photometric multi-pass filter do not overlap each other;
Wherein, all pass bands of the luminous multi-pass filter are not overlapped with each other, and all pass bands of the photometric multi-pass filter are not overlapped with each other;
Wherein one passband of the dichroic mirror corresponds to one passband of the photometric multi-pass filter, and each passband of the luminescent multi-pass filter and each passband of the dichroic mirror do not overlap each other;
wherein all pass bands of the dichroic mirror do not overlap each other;
The test tube is internally provided with a plurality of different dyes and a tested target, and the fluorescence detection intensity of each dye is positively correlated with the concentration of the tested target;
One passband of the luminescent multi-pass filter corresponds to the excitation wavelength of one of the dyes; one passband of the photometric multi-pass filter corresponds to the fluorescence wavelength of one of the dyes.
6. The detection method according to claim 5, wherein a light-emitting lens is disposed between the multicolor light-emitting element and the light-emitting multi-pass filter, for transmitting excitation light of different spectral ranges emitted by the multicolor light-emitting element to the light-emitting multi-pass filter;
and a light measuring lens is arranged between the light multi-pass filter and the light measuring element and is used for transmitting fluorescence with different wavelengths separated by the light measuring multi-pass filter to the light measuring element.
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