CN213337348U

CN213337348U - Micro-droplet double-fluorescence signal detection device

Info

Publication number: CN213337348U
Application number: CN202021932719.6U
Authority: CN
Inventors: 黄海旺; 夏雷; 于海侠; 周跃; 梁欢迎; 白宇; 杨文军
Original assignee: Xinyi Manufacturing Technology Beijing Co ltd
Current assignee: Xinyi Manufacturing Technology Beijing Co ltd
Priority date: 2020-09-07
Filing date: 2020-09-07
Publication date: 2021-06-01
Anticipated expiration: 2030-09-07

Abstract

The utility model relates to a two fluorescence signal detection device of micro-droplet includes: the device comprises a light combination module, an objective lens and a light processing module. The light management module also includes a mounting housing configured with a light conducting channel including a single aperture corresponding to both the first photomultiplier tube and the second photomultiplier tube. The utility model discloses a single detects and can acquire multinomial detection parameter to form the light processing framework that single aperture copolymerization is burnt, two detect the light path and use common aperture, reach two tunnel emission luminous efficiency increase of equal proportion or reduce, save manufacturing cost.

Description

Micro-droplet double-fluorescence signal detection device

Technical Field

The utility model relates to a micro-fluidic chip technical field, concretely relates to two fluorescence signal detection devices of micro-droplet.

Background

The biochip has wide application in new medicine development, disease diagnosis, gene expression analysis, etc. The technology of the micro-fluidic chip is mature day by day and becomes a focus of people. There are various biological and chemical processes in the microfluidic detection chip, and the processes are usually completed in the micro-scale flow channel space, wherein some devices capable of detecting the reaction process are also required. The existing detection means can be divided into CCD scanning and laser confocal scanning, the CCD scanning system has a simpler structure and a higher detection speed compared with the laser confocal scanning system, but has a lower transverse resolution, if the transverse resolution is required to be improved, the amplification factor of the imaging system needs to be improved, and the corresponding field of view is reduced (namely the area of a chip to be measured at one time is smaller), when the area of the chip to be measured is larger, the chip can be spliced after being measured in blocks for many times, and because the block scanning actually makes the chip and the imaging system move relatively in a mechanical movement mode, the mechanical positioning error forms the splicing error of a scanned image, so the method is not suitable for high-precision high-density biochip detection.

The biochip detection system constructed based on the laser confocal principle scans the biochip point by point, and the biochip is always positioned on a focal plane, so that the spot size of the exciting light is very small, and the transverse resolution is high. Because the laser confocal detection mode has the characteristics of high resolution and high sensitivity, clear digital fluorescence images and quantitative analysis results of antibodies and the like marked by fluorescence on the biochip can be obtained, and the method can become a new detection method mainly adopted by high-density biochip scanning. When the two PMTs use the same small hole to shield stray light, the light emitting efficiency of the two PMTs is increased or reduced in equal proportion, and the purpose of unifying crosstalk correction parameters is achieved.

SUMMERY OF THE UTILITY MODEL

Therefore, the to-be-solved technical problem of the utility model is to provide a two fluorescence signal detection devices of micro-droplet can acquire the two fluorescence detection signals of micro-droplet through closing optical module and light processing module to form the light processing framework that single aperture is confocal, two detection light paths use same single aperture, reach two tunnel emission luminous efficiency equal proportion increase or reduce, save manufacturing cost.

In order to solve the above problem, the utility model provides a two fluorescence signal detection devices of micro-droplet, include:

the light combining module is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed exciting light;

the objective lens is positioned on a light conduction path of the mixed exciting light and is used for confocal focusing the mixed exciting light on the micro-droplet to be detected so as to excite and generate first fluorescence corresponding to the first laser and second fluorescence corresponding to the second laser;

the light processing module comprises a first photomultiplier for acquiring first fluorescence, a second photomultiplier for acquiring second fluorescence, a first dichroic mirror for reflecting the mixed excitation light to the objective lens from the light combining module, and a second dichroic mirror for separating the first fluorescence from the second fluorescence;

the light management module further includes a mounting housing configured with a light conduction channel including a single aperture corresponding to both the first photomultiplier tube and the second photomultiplier tube.

Preferably, the light processing module further includes a convex lens between the first dichroic mirror and the second dichroic mirror, the convex lens being configured to focus parallel light passing therethrough.

Preferably, the aperture is disposed between the convex lens and the second dichroic mirror so that light passing through the convex lens is focused on the aperture.

Preferably, the light processing module further comprises a first color filter between the first photomultiplier tube and the second dichroic mirror, and/or a second color filter between the second photomultiplier tube and the second dichroic mirror.

Preferably, the mounting housing includes a fixing member, a first connecting member, a second connecting member, a third connecting member, and a fourth connecting member, the fixing member has a first light ray injection opening and a first light ray injection opening, the first light ray injection opening is coaxial with the first light ray injection opening, and a small hole is disposed in the first light ray injection opening; the third connecting piece is connected to the first light emitting outlet of the fixing piece, and is provided with a second light emitting inlet, a second light emitting outlet and a third light emitting outlet, wherein the second light emitting inlet is coaxial with the first light emitting outlet of the fixing piece, the second light emitting outlet is coaxial with the second light emitting inlet, and the third light emitting outlet is perpendicular to the second light emitting inlet; the second connecting piece is connected to the first light ray incidence opening of the fixing piece, and the convex lens is clamped between the second connecting piece and the first light ray incidence opening of the fixing piece; the fourth connecting piece is connected to the second light ray emitting opening of the third connecting piece, the second dichroic mirror is clamped between the fourth connecting piece and the second light ray emitting opening of the third connecting piece, a first photomultiplier is arranged at one end of the fourth connecting piece, which is far away from the fixing piece, and the first color filter is clamped between the fourth connecting piece and the first photomultiplier along a light ray conducting path; a third light ray outlet of the third connecting piece is connected with a second photomultiplier, and a second color filter is clamped between the third connecting piece and the second photomultiplier fixing seat along a light ray conducting path; the first connecting piece is connected to one side, far away from the fixing piece, of the second connecting piece, and the first dichroic mirror is clamped between the first connecting piece and the second connecting piece.

Preferably, a joint between the first connecting piece and the second connecting piece, and/or a joint between the second connecting piece and the fixing piece, and/or a joint between the third connecting piece and the fourth connecting piece, and/or a joint between the third connecting piece and the fixing piece, and/or a joint between the first photomultiplier and the fourth connecting piece, and/or a joint between the second photomultiplier and the third connecting piece is provided with a sealing shock-absorbing piece.

Preferably, the objective lens is movably connected to a side of the first connecting piece, which is away from the second connecting piece.

Preferably, the light combining module includes a first laser, a second laser, a reflective mirror and a third dichroic mirror, and the first laser light emitted by the first laser is reflected by the reflective mirror and then combined with the second laser light emitted by the second laser into the mixed excitation light at the third dichroic mirror.

Preferably, the light combining module further includes a fixing plate and a light combining cassette, the first laser, the second laser and the light combining cassette are fixedly connected to the fixing plate, the reflective mirror and the third dichroic mirror are disposed in the light combining cassette, and the light combining cassette has a mixed excitation light exit port.

Preferably, the first laser is a 532nm laser; and/or the second laser is a 473nm laser.

The utility model provides a pair of two fluorescence signal detection device of micro-droplet is synthesized through the first laser and the second laser of wavelength difference and is formed treat behind the mixed exciting light and detect the micro-droplet and arouse and form first fluorescence and second fluorescence, and pass through first photomultiplier and second photomultiplier reach after acquireing respectively and carry out corresponding processing (for example count) to corresponding data processing equipment (such as computer) to realized that the single detects and to acquire the multinomial detection parameter, and form the light processing framework that single aperture copolymerization is burnt, two detection light paths use common aperture, reach two tunnel emission luminous efficiency equal proportion increase or reduce, save manufacturing cost.

Drawings

Fig. 1 is a schematic view of an exploded structure of a micro-droplet dual fluorescence signal detection device according to an embodiment of the present invention;

fig. 2 is a schematic diagram of a light transmission path of the micro-droplet dual fluorescence signal detection device according to an embodiment of the present invention;

FIG. 3 is an exploded view of the optical processing module of FIG. 1;

fig. 4 is an exploded structural diagram of the light combining module in fig. 1.

The reference signs are:

1. a light combining module; 11. a first laser; 12. a second laser; 13. a fixing plate; 14. a light-combining cassette; 141. a reflective mirror; 142. a third dichroic mirror; 2. an objective lens; 3. A light processing module; 31. a first photomultiplier tube; 32. a second photomultiplier tube; 33. a first dichroic mirror; 34. a second dichroic mirror; 35. a first color filter; 36. a second color filter; 371. a fixing member; 372. a first connecting member; 373. a second connecting member; 374. a fourth connecting member; 375. a third connecting member; 376. a small hole; 38. a convex lens; 391. a shock pad; 392. a gasket; 393. a light-tight pad; 100. micro-droplets are detected.

Detailed Description

Referring to fig. 1 to 4 in combination, according to an embodiment of the present invention, there is provided a micro-droplet dual fluorescence signal detection apparatus, including: the light combining module 1 is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed excitation light; the objective lens 2 is located on a light conduction path of the mixed excitation light, and is used for confocal focusing the mixed excitation light on the microdroplet 100 to be detected so as to excite and generate first fluorescence corresponding to the first laser and second fluorescence corresponding to the second laser; the light processing module 3 includes a first photomultiplier 31 for obtaining the first fluorescence, a second photomultiplier 32 for obtaining the second fluorescence, a first dichroic mirror 33 for reflecting the mixed excitation light from the light combining module to the objective lens 2, and a second dichroic mirror 34 for separating the first fluorescence from the second fluorescence. In the technical scheme, the mixed excitation light is formed by synthesizing the first laser and the second laser with different wavelengths, then the micro-droplet 100 to be detected is excited to form the first fluorescence and the second fluorescence, the obtained mixed excitation light is respectively transmitted to corresponding data processing equipment (such as a computer and the like) through the first photomultiplier tube 31 and the second photomultiplier tube 32 to be correspondingly processed (such as counting), so that multiple detection parameters can be obtained by single detection, a single-pore confocal light processing framework is formed, two detection light paths use a common pore, the equal proportion increase or decrease of the efficiency of two paths of emitted light is achieved, and the production cost is saved. It is worth mentioning that, because the detection device of this application forms the light processing framework of single aperture confocal, when two detection light paths used same aperture shielding stray light, two tunnel transmitting light efficiency equal proportion increase or reduce reached the purpose of unified crosstalk correction parameter, the structure still less simultaneously, the debugging process is simpler, very big improvement production efficiency.

Preferably, the light processing module 3 further includes a convex lens 38, and the convex lens 38 is located between the first dichroic mirror 33 and the second dichroic mirror 34, so that parallel light passing through the convex lens is focused to the small hole 376, so that external interference light is shielded by the small hole 376, and the detection accuracy of subsequent fluorescent signals is ensured.

Further, the light processing module 3 further includes a first color filter 35, the first color filter 35 being located between the first photomultiplier tube 31 and the second dichroic mirror 34, and/or the light processing module 3 further includes a second color filter 36, the second color filter 36 being located between the second photomultiplier tube 32 and the second dichroic mirror 34.

Specifically, the light processing module 3 further has a mounting housing configured with a light conducting channel having apertures 376 corresponding to the first photomultiplier tube 31 and the second photomultiplier tube 32. More specifically, the mounting housing includes a fixing member 371, a first connecting member 372, a second connecting member 373, a third connecting member 375, and a fourth connecting member 374, wherein the fixing member 371 has a first light incident opening and a first light exiting opening, the first light incident opening and the first light exiting opening are coaxial, and a small hole 376 is disposed in the first light exiting opening; the third connecting member 375 is connected to the first light emitting opening of the fixing member 371, the third connecting member 375 has a second light emitting opening, a second light emitting opening and a third light emitting opening, the second light emitting opening is coaxial with the first light emitting opening of the fixing member 371, the second light emitting opening is coaxial with the second light emitting opening, and the third light emitting opening is perpendicular to the second light emitting opening; the second connecting member 373 is connected to the first light incident port of the fixing member 371, and the convex lens 38 is sandwiched therebetween; the fourth connecting member 374 is connected to the second light emitting port of the third connecting member 375, and the second dichroic mirror 34 is interposed therebetween, a first photomultiplier tube 31 is disposed at one end of the fourth connecting member 374 away from the fixing member 371, and the first color filter 35 is interposed between the fourth connecting member 374 and the first photomultiplier tube 31 along a light conducting path; the third connecting member 375 is connected to the second photomultiplier tube 32 at the third light exit, and the second color filter 36 is sandwiched between the third connecting member 375 and the second photomultiplier tube 32 along the light conducting path; the first connecting member 372 is connected to a side of the second connecting member 373 away from the fixing member 371, and the first dichroic mirror 33 is sandwiched between the first connecting member and the second connecting member. In this technical solution, a specific implementation manner of the light processing module 3 is provided, which is formed by mutually assembling and assembling each relatively independent connecting component and parts, and it can be understood that, in order to ensure the rationality of the light conducting channel, in particular, the design of the light conducting channel is in accordance with the light processing path shown in fig. 2 of the present application, that is, although the specific definition of the corresponding light conducting channels in the first connecting component 372, the second connecting component 373, the third connecting component 375, and the fourth connecting component 374 is not provided in the foregoing, it is clear that, in order to ensure the light conducting requirements of fig. 2 and the compactness of the overall structure of the light processing module 3, the first connecting component 372, the second connecting component 373, the fixing component 371, the third connecting component 375, and the fourth connecting component 374 are sequentially located on the light conducting path and satisfy the light conducting requirements shown in fig. 2, the first connector 372, the second connector 373, the third connector 375, and the fourth connector 374 all adopt isosceles right triangles in shape, that is, they all have an inclined plane of 45 °.

Further, in order to protect the optical lens such as the convex lens 38 disposed in the light processing module 3 from being damaged and ensure that the optical lens is protected from the external light, it is preferable that the connection between the first connection member 372 and the second connection member 373, and/or the connection between the second connection member 373 and the fixing member 371, and/or the connection between the third connection member 375 and the fourth connection member 374, and/or the connection between the third connection member 375 and the fixing member 371, and/or the connection between the first photomultiplier tube 31 and the fourth connection member 374, and/or the connection between the second photomultiplier tube 32 and the third connection member 375 are disposed with a sealing shock absorber, which may include a shock absorbing pad 391, a sealing gasket 392, a sealing washer 392, and a sealing shock absorbing member, One or more of the gaskets 393.

The objective lens 2 is movably connected to a side of the first connecting member 372 away from the second connecting member 373 as a component for performing focus detection on the microdroplets 100 to be detected, as a specific embodiment, it is only required to use a 20-fold microscope, and it can be understood that the movable connection here means that the objective lens 2 has a rotational (around Z axis) degree of freedom and a translational (along Z axis or X axis) degree of freedom, so as to ensure that a detection person can flexibly adjust a focal point (focal plane) of the objective lens 2, and further, it can be understood that the aforementioned movement of the objective lens 2 can be controlled by a power module such as a lead screw motor.

As one specific embodiment of the light combining module 1, preferably, the light combining module 1 includes a first laser 11, a second laser 12, a reflective mirror 141, and a third dichroic mirror 142, and the first laser light emitted by the first laser 11 is reflected by the reflective mirror 141 and then combined with the second laser light emitted by the second laser 12 into the mixed excitation light by the third dichroic mirror 142. For example, the first laser 11 is a 532nm laser, in which case it is a green laser; and/or the second laser 12 is a 473nm laser, in this case a blue laser. More specifically, the light combining module 1 further includes a fixing plate 13 and a light combining cassette 14, the first laser 11, the second laser 12 and the light combining cassette 14 are fixedly connected to the fixing plate 13, the reflective mirror 141 and the third dichroic mirror 142 are disposed in the light combining cassette 14, and the light combining cassette 14 has a mixed excitation light exit. At this time, it can be understood that the light combining module 1 is constructed as a whole, so that the micro-droplet dual fluorescence signal detection apparatus is further compact in structure.

It is readily understood by a person skilled in the art that the advantageous ways described above can be freely combined, superimposed without conflict.

The above description is only exemplary of the present invention and should not be construed as limiting the present invention, and any modifications, equivalents and improvements made within the spirit and principles of the present invention are intended to be included within the scope of the present invention. The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims

1. A micro-droplet dual fluorescence signal detection device, the detection device comprising:

the light combining module (1) is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed excitation light;

the objective lens (2) is positioned on a light conduction path of the mixed exciting light and is used for focusing the mixed exciting light on the micro-droplet (100) to be detected so as to excite and generate first fluorescence corresponding to the first laser and second fluorescence corresponding to the second laser;

light processing module (3) comprising a first photomultiplier (31) to obtain a first fluorescence, a second photomultiplier (32) to obtain a second fluorescence, a first dichroic mirror (33) to reflect the mixed excitation light from the light combining module to the objective (2), and a second dichroic mirror (34) to separate the first and second fluorescence, said light processing module (3) further comprising a mounting housing configured with a light conducting channel comprising a single aperture (376) corresponding to both the first photomultiplier (31) and the second photomultiplier (32).

2. The detection device according to claim 1, characterized in that the light processing module (3) further comprises a convex lens (38), the convex lens (38) being between the first dichroic mirror (33) and the second dichroic mirror (34), the convex lens (38) being configured so as to focus parallel light passing through it.

3. The detection apparatus according to claim 2, wherein the aperture (376) is arranged between the convex lens (38) and the second dichroic mirror (34) such that light passing through the convex lens (38) is focused at the aperture.

4. The detection apparatus according to claim 2, wherein the light processing module (3) further comprises a first color filter (35), the first color filter (35) being between the first photomultiplier tube (31) and the second dichroic mirror (34), and/or wherein the light processing module (3) further comprises a second color filter (36), the second color filter (36) being between the second photomultiplier tube (32) and the second dichroic mirror (34).

5. The detecting device according to claim 4, wherein the mounting housing includes a fixing member (371), a first connecting member (372), a second connecting member (373), a third connecting member (375), and a fourth connecting member (374), the fixing member (371) has a first light injecting opening and a first light emitting opening, the first light injecting opening is coaxial with the first light emitting opening, and the small hole (376) is disposed at the first light emitting opening; the third connecting member (375) is connected to the first light emitting opening of the fixing member (371), the third connecting member (375) has a second light emitting opening, and a third light emitting opening, the second light emitting opening is coaxial with the first light emitting opening of the fixing member (371), the second light emitting opening is coaxial with the second light emitting opening, and the third light emitting opening is perpendicular to the second light emitting opening; the second connecting piece (373) is connected to the first light ray incidence port of the fixing piece (371), and the convex lens (38) is clamped between the second connecting piece and the first light ray incidence port; the fourth connecting piece (374) is connected to the second light emitting outlet of the third connecting piece (375), the second dichroic mirror (34) is clamped between the fourth connecting piece and the second light emitting outlet of the third connecting piece (375), a first photomultiplier (31) is arranged at one end of the fourth connecting piece (374) departing from the fixing piece (371), and the first color filter (35) is clamped between the fourth connecting piece (374) and the first photomultiplier (31) along a light conducting path; a second photomultiplier is connected to the third light emitting outlet of the third connecting piece (375), and the second color filter (36) is clamped between the third connecting piece (375) and the second photomultiplier (32) along a light conducting path; the first connecting piece (372) is connected to one side, away from the fixing piece (371), of the second connecting piece (373), and the first dichroic mirror (33) is clamped between the first connecting piece and the second connecting piece.

6. The detection apparatus according to claim 5, wherein a junction between the first connection member (372) and the second connection member (373), and/or a junction between the second connection member (373) and the fixing member (371), and/or a junction between the third connection member (375) and the fourth connection member (374), and/or a junction between the third connection member (375) and the fixing member (371), and/or a junction between the first photomultiplier tube (31) and the fourth connection member (374), and/or a junction between the second photomultiplier tube (32) and the third connection member (375) is provided with a sealing shock-absorbing member.

7. The detection apparatus according to claim 5, wherein the objective lens (2) is movably connected to a side of the first connection member (372) facing away from the second connection member (373).

8. The detection device according to any one of claims 1 to 7, wherein the light combining module (1) comprises a first laser (11), a second laser (12), a reflective mirror (141), and a third dichroic mirror (142), and the first laser light emitted from the first laser (11) is reflected by the reflective mirror (141) and combined with the second laser light emitted from the second laser (12) into the mixed excitation light at the third dichroic mirror (142).

9. The detecting device according to claim 8, wherein the light combining module (1) further comprises a fixing plate (13) and a light combining cassette (14), the first laser (11), the second laser (12) and the light combining cassette (14) are fixedly connected to the fixing plate (13), the reflective mirror (141) and the third dichroic mirror (142) are disposed in the light combining cassette (14), and the light combining cassette (14) has a mixed excitation light exit.

10. Detection apparatus according to claim 8, characterized in that the first laser (11) is a 532nm laser; and/or the second laser (12) is a 473nm laser.