CN114806842A - Digital PCR fluorescence detection device - Google Patents

Abstract

The invention relates to a digital PCR fluorescence detection device, which comprises an excitation light source module, a filtering module, a carrying module and an imaging module, wherein the excitation light source module comprises a mounting seat, a dichroic mirror and a plurality of monochromatic light sources, an exit port is formed in one end of the mounting seat, the plurality of monochromatic light sources are mutually parallel or vertically arranged at the other end of the mounting seat, the dichroic mirror and the monochromatic light sources are arranged at an angle of 45 degrees, and light rays of the plurality of monochromatic light sources are transmitted or reflected to the exit port through the dichroic mirror to be emitted; the filtering module comprises a base and a turntable which is rotationally connected with the base, and the turntable is provided with a plurality of filtering components; the carrying module is arranged on one side of the base, and the imaging module is arranged on the other side of the base. The whole structure is compact and simple, the volume is small, and a certain propulsion effect is realized on the real-time on-site detection; and a plurality of circumferentially arranged filtering assemblies are arranged through the rotary disc, so that the detection flux is improved.

Description

Digital PCR fluorescence detection device

Technical Field

The invention relates to the technical field of medical detection, in particular to a digital PCR fluorescence detection device.

Background

There are two main methods for fluorescence detection of droplet digital PCR (Polymerase Chain Reaction): one method is to adopt a flow cytometry method, use a microfluidic technology to enable micro-droplets to sequentially pass through a bicolor optical detection system at a high speed, and use a photoelectric detector and a filter to collect excited fluorescent signals. The other method is an imaging statistical method, a high-brightness light source is adopted to irradiate a microfluidic chip, then a fluorescence microscope or a cold CCD is used for collecting fluorescence signals, positive droplets and negative droplets are counted, the counting is carried out on the positive droplets based on an image processing method, generally, a high-power white light LED or a mercury lamp is used as the light source, the spectral range is wide, in order to prevent fluorescence crosstalk, the bandwidths of an excitation filter and an emission filter are narrow, and the light energy utilization rate is low.

Aiming at an imaging statistical method, in order to solve the problem that a high-power white light LED or mercury lamp is used as a light source but the light energy utilization rate is not high, a monochromatic light source can be adopted, but in order to meet the requirement of practical application, two or more than two fluorescent dyes are usually used, which requires that two or more than two monochromatic light sources and optical elements matched with the monochromatic light sources are configured; however, the general droplet-type digital PCR equipment is huge in size, limited by the imaging field of view and inconvenient for realizing real-time field detection.

Disclosure of Invention

In view of the above, it is necessary to provide a digital PCR fluorescence detection apparatus to solve the problem of the large volume of the droplet-type digital PCR device.

A digital PCR fluorescence detection device comprises

The excitation light source module comprises a mounting seat, a dichroic mirror and a plurality of monochromatic light sources, wherein an exit port is formed in one end of the mounting seat, the monochromatic light sources are arranged at the other end of the mounting seat in parallel or vertically, the dichroic mirror and the monochromatic light sources are arranged at an angle of 45 degrees, and light rays of the monochromatic light sources are transmitted or reflected to the exit port to be emitted through the dichroic mirror;

the filtering module comprises a base and a turntable which is rotationally connected with the base, the turntable is provided with a plurality of filtering components along the circumferential direction of the turntable, the base is provided with a first opening, a second opening and a third opening, and the first opening corresponds to the emergent port;

the carrying module is arranged on one side of the base and used for carrying a chip to be detected, and the second opening faces the carrying module;

and the imaging module is arranged on the other side of the base, and the third opening faces the imaging module.

In one embodiment, the digital PCR fluorescence detection apparatus further includes a collimating lens disposed on the mounting seat, the plurality of monochromatic light sources include a first monochromatic light source and a second monochromatic light source, the dichroic mirror includes a first dichroic mirror and a second dichroic mirror, the first monochromatic light source and the second monochromatic light source are perpendicular to each other and are axially symmetric with respect to the first dichroic mirror, the first dichroic mirror and the second dichroic mirror are perpendicular to each other, and the first dichroic mirror, the second dichroic mirror and the collimating lens are sequentially disposed along an optical axis direction of the first monochromatic light source.

In one embodiment, the excitation light source module further includes a third monochromatic light source, a fourth monochromatic light source, and a third dichroic mirror, which are mounted on the mounting seat, the third monochromatic light source and the fourth monochromatic light source are perpendicular to each other and are axially symmetric with respect to the third dichroic mirror, the third dichroic mirror and the second dichroic mirror are parallel to each other, an extension line of the first monochromatic light source is perpendicular to an extension line of the fourth monochromatic light source, the second monochromatic light source is perpendicular to an extension line of the third monochromatic light source, distances from midpoints of the first monochromatic light source and the fourth monochromatic light source to the second dichroic mirror are equal, and the first monochromatic light source and the fourth monochromatic light source are axially symmetric with respect to the second dichroic mirror.

In one embodiment, the excitation light source module further includes a double cemented lens mounted on the mounting base, and the double cemented lens and the collimating lens are disposed along the same optical axis direction and located downstream of the collimating lens.

In one embodiment, the excitation light source module further includes a fly-eye lens mounted on the mounting base, the fly-eye lens and the collimating lens are arranged along the same optical axis direction, and the fly-eye lens is located between the collimating lens and the double-cemented lens.

In one embodiment, the filter assembly comprises a fixing seat, an excitation filter, a fourth dichroic mirror and an emission filter, wherein the excitation filter, the fourth dichroic mirror and the emission filter are mounted on the fixing seat, the excitation filter and the emission filter are perpendicular to each other, the fourth dichroic mirror and the excitation filter reach 45 degrees respectively, a fourth opening is arranged on the side face, opposite to the emission filter, of the fixing seat, when the rotating disc rotates to a preset position, the excitation filter is opposite to the first opening, the fourth opening is opposite to the second opening, and the emission filter is opposite to the third opening.

In one embodiment, the base includes a bottom case and an upper cover connected to the bottom case, the bottom case and the upper cover enclose an accommodating cavity for accommodating the turntable and the filter assembly, and the turntable is rotatably connected to the bottom case and/or the upper cover.

In one embodiment, the base further comprises a sealing cover, the upper cover is provided with an operation window communicated with the accommodating cavity, and the sealing cover is rotatably connected with the upper cover so as to open or close the operation window;

and/or the upper cover is provided with a clamping bead, and the clamping bead partially protrudes into the accommodating cavity so that the fixing seat is limited.

In one embodiment, the carrier module includes a first support frame, a first sliding rod, a second support frame, a second sliding rod, and a chip rack, wherein the first support frame is fixed on the base, the first sliding rod is fixed on the first support frame and located above the base, the second support frame is slidably connected with the first sliding rod, the second sliding rod is fixed on the second support frame, the chip rack is slidably connected with the second sliding rod, and the first sliding rod and the second sliding rod are perpendicular to each other;

and/or, the imaging module comprises a lens and an image sensor connected with the lens, and the lens is fixed at the lower end of the base.

In one embodiment, the digital PCR fluorescence detection apparatus further includes a support rod and a fixing plate, one end of the support rod is connected to the base, the other end of the support rod is connected to the fixing plate, and the length of the support rod is greater than the length of the imaging module.

The digital PCR fluorescence detection device is characterized in that a chip to be detected is arranged on a carrying module, a certain monochromatic light source is selected according to the difference of fluorescent dyes on the chip, a filter component corresponding to the light source is selected by correspondingly rotating a turntable, light of the monochromatic light source penetrates through a dichroic mirror or is reflected on the surface of the dichroic mirror and emitted from an exit port, the light enters the filter module through a first opening, the light processed by the filter component is emitted from a second opening and irradiates on the chip, droplets are filled in the chip, a fluorescence signal with a wavelength slightly larger than that of exciting light (a certain monochromatic light source) is emitted due to Stokes displacement under the irradiation of the exciting light, and the fluorescence signal penetrates through the filter component and is finally captured by an imaging module through a third opening. By analyzing the pictures taken by the imaging module, counts of positive or negative droplets can be obtained. According to the digital PCR fluorescence detection device, the mounting base is provided with the plurality of monochromatic light sources, the monochromatic light sources are emitted to the filtering module through the shared dichroic mirror, the plurality of filtering assemblies are arranged on the turntable along the circumferential direction of the turntable and can be matched with the corresponding monochromatic light sources, the carrying modules and the imaging modules are arranged on two sides of the base of the filtering module to realize chip positioning and imaging, the whole structure is compact and simple, the size is small, and a certain propulsion effect is realized on real-time on-site detection; establish a plurality of filter assembly that circumference was arranged through the carousel, when increasing monochromatic light source according to the demand, can correspond the quantity that increases filter assembly, improve and detect the flux, shorten check-out time.

Drawings

FIG. 1 is a schematic perspective view of a digital PCR fluorescence detection apparatus according to an embodiment;

FIG. 2 is a schematic top view of an excitation light source module of the digital PCR fluorescence detection apparatus according to an embodiment;

FIG. 3 is a schematic diagram illustrating a filter module of the digital PCR fluorescence detection apparatus according to an embodiment of the present invention;

fig. 4 is an assembly view of the filter module shown in fig. 3.

Description of reference numerals:

10. an excitation light source module; 110. a mounting seat; 101. an exit port; 111. a first monochromatic light source; 112. a second monochromatic light source; 113. a third monochromatic light source; 114. a fourth monochromatic light source; 121. a first dichroic mirror; 122. a second dichroic mirror; 123. a third dichroic mirror; 13. a collimating lens; 14. a double cemented lens; 15. a fly-eye lens; 20. a filtering module; 210. a base; 201. a first opening; 202. a second opening; 212. a bottom case; 214. an upper cover; 216. sealing the cover; 218. an operating window; 219. clamping the beads; 220. a turntable; 230. a filtering component; 232. a fixed seat; 234. exciting the filter plate; 236. a fourth dichroic mirror; 238. an emission filter; 30. a carrier module; 310. a first support frame; 320. a first slide bar; 330. a second support frame; 340. a second slide bar; 350. a chip holder; 40. an imaging module; 410. a lens; 420. an image sensor; 430. a connecting member; 50. a support bar; 60. a fixing plate; 70. and (3) a chip.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Referring to fig. 1, an embodiment of a digital PCR fluorescence detection apparatus includes an excitation light source module 10, a filtering module 20, a carrier module 30, and an imaging module 40. Referring to fig. 2, in one embodiment, the excitation light source module 10 includes a mounting base 110, a dichroic mirror and a plurality of monochromatic light sources, an exit port 101 is disposed at one end of the mounting base 110, the plurality of monochromatic light sources are disposed at the other end of the mounting base 110 in parallel or perpendicular to each other, the dichroic mirror and the monochromatic light sources are disposed at an angle of 45 °, and light rays of the plurality of monochromatic light sources are transmitted or reflected through the dichroic mirror to the exit port 101 for exit.

Alternatively, the monochromatic light source may include at least two of a blue light source, a green light source, a red light source, a violet light source, a yellow light source, or the like. The dichroic mirror reflects light below the target wavelength and transmits light above the target wavelength. Referring to fig. 1, a rectangular bump may be disposed on a wall surface of the mounting base 110, a light-transmitting hole is formed on the wall surface of the rectangular bump, the monochromatic light source is fixed on the surface of the rectangular bump, and light emitted from the monochromatic light source enters the mounting base 110 through the light-transmitting hole and is then emitted toward the dichroic mirror.

In one embodiment, the filtering module 20 includes a base 210 and a turntable 220 rotatably connected to the base 210, the turntable 220 is provided with a plurality of filtering components 230 along a circumferential direction thereof, the base 210 is provided with a first opening 201, a second opening 202 and a third opening, and the first opening 201 corresponds to the exit 101. Different fluorescent dyes are correspondingly matched with different filtering assemblies 230 and monochromatic light sources with different wavelengths, the number of the filtering assemblies 230 arranged on the turntable 220 is not less than that of the monochromatic light sources, the detection flux is improved, the detection time is shortened, the turntable 220 rotates relative to the base 210, so that the corresponding filtering assemblies 230 rotate to correspond to the first opening 201, the second opening 202 and the third opening, and the treatment of exciting light and autofluorescence is realized. In one embodiment, the carrier module 30 is disposed on one side of the base 210 for carrying the chip 70 to be tested, and the second opening 202 faces the carrier module 30. In one embodiment, the imaging module 40 is disposed on the other side of the base 210, and the third opening faces the imaging module 40.

When the digital PCR fluorescence detection apparatus of the above embodiment is used, the chip 70 to be detected is placed in the object-carrying module 30, a certain monochromatic light source is selected according to the difference of fluorescent dyes on the chip 70, the filter component 230 corresponding to the light source is selected corresponding to the rotating disc 220, light of the monochromatic light source is emitted from the exit port 101 after passing through the dichroic mirror or being reflected on the surface of the dichroic mirror, enters the filter module 20 through the first opening 201, the light processed by the filter component 230 is emitted from the second opening 202 and irradiated onto the chip 70, droplets are filled in the chip 70, a fluorescent signal with a wavelength slightly larger than that of excitation light (a certain monochromatic light source) is emitted due to stokes shift under the irradiation of the excitation light, and the fluorescent signal passes through the filter component 230 and is finally captured by the imaging module 40 through the third opening. By analyzing the photographs taken by the imaging module 40, counts of positive or negative droplets can be obtained. According to the digital PCR fluorescence detection device, a plurality of high-power monochromatic light sources with different wavelengths are configured on the mounting seat 110, dichroic mirrors are set according to different wave bands and are emitted to the filtering module 20 through the shared dichroic mirrors, the plurality of filtering components 230 are arranged on the rotary table 220 along the circumferential direction of the rotary table and can be matched with the corresponding monochromatic light sources, the carrying modules 30 and the imaging modules 40 are arranged on two sides of the base 210 of the filtering module 20 to achieve positioning and imaging of the chip 70, the whole structure is compact and simple, the size is small, and a certain propelling effect is achieved for real-time on-site detection; establish a plurality of filter assembly 230 of circumference arrangement through carousel 220, when increasing monochromatic light source according to the demand, can correspond the quantity that increases filter assembly 230, improve and detect the flux, shorten the check-out time.

Further, in one embodiment, the digital PCR fluorescence detection device further includes a collimating lens 13 disposed on the mounting base 110. The light passes through the dichroic mirror and then vertically passes through the collimating lens 13, and the light beam passes through the collimating lens 13 and then becomes parallel light to be emitted.

Referring to fig. 2, optionally, in an embodiment, the plurality of monochromatic light sources includes a first monochromatic light source 111 and a second monochromatic light source 112, and the dichroic mirror includes a first dichroic mirror 121 and a second dichroic mirror 122. The first monochromatic light source 111 and the second monochromatic light source 112 are perpendicular to each other and are axisymmetric with respect to the first dichroic mirror 121. The first dichroic mirror 121 and the second dichroic mirror 122 are perpendicular to each other. The first dichroic mirror 121, the second dichroic mirror 122, and the collimator lens 13 are sequentially arranged in the optical axis direction of the first monochromatic light source 111. The light emitted from the first monochromatic light source 111 passes through the first dichroic mirror 121, the second dichroic mirror 122, and the collimating lens 13 in sequence, and then is emitted from the exit port 101 to the filtering module 20. The light emitted from the second monochromatic light source 112 is reflected by the surface of the first dichroic mirror 121, and then sequentially passes through the second dichroic mirror 122 and the collimating lens 13, and then is emitted to the filtering module 20 through the exit port 101. The first monochromatic light source 111 and the second monochromatic light source 112 can be disposed on different walls of the same rectangular bump.

Referring to fig. 2, optionally, in one embodiment, the excitation light source module 10 further includes a third monochromatic light source 113, a fourth monochromatic light source 114 and a third dichroic mirror 123 mounted on the mounting base 110. The third monochromatic light source 113 and the fourth monochromatic light source 114 are perpendicular to each other and are axisymmetric with respect to the third dichroic mirror 123. The third dichroic mirror 123 and the second dichroic mirror 122 are parallel to each other. An extension line of the first monochromatic light source 111 is perpendicular to an extension line of the fourth monochromatic light source 114, and the second monochromatic light source 112 is perpendicular to an extension line of the third monochromatic light source 113. The midpoints of first monochromatic light source 111 and fourth monochromatic light source 114 are equidistant from second dichroic mirror 122. First monochromatic light source 111 and fourth monochromatic light source 114 are axisymmetric with respect to second dichroic mirror 122. The light emitted from the third monochromatic light source 113 is reflected by the surface of the third dichroic mirror 123, then emitted to the surface of the second dichroic mirror 122, reflected by the surface of the second dichroic mirror 122, and then emitted from the exit port 101 to the filter module 20 through the collimating lens 13. The light emitted from the fourth monochromatic light source 114 passes through the third dichroic mirror 123, then is emitted to the surface of the second dichroic mirror 122, is reflected by the surface of the second dichroic mirror 122, and then is emitted from the exit port 101 to the filter module 20 through the collimating lens 13. The third monochromatic light source 113 and the fourth monochromatic light source 114 can be disposed on different walls of the same rectangular bump. In order to satisfy the requirement that the excitation light spots acquired by selecting different channels are uniform and equally large, the distance from each monochromatic light source to the chip 70 satisfies the aplanatic condition.

Optionally, in other embodiments, two or three of the first monochromatic light source 111, the second monochromatic light source 112, the third monochromatic light source 113, and the fourth monochromatic light source 114 may also be selected for combination, such as only the third monochromatic light source 113 and the fourth monochromatic light source 114 are provided; or only the first monochromatic light source 111 and the third monochromatic light source 113 are provided; or only the first monochromatic light source 111 and the fourth monochromatic light source 114 may be provided.

Further, in one embodiment, the excitation light source module 10 further includes a double cemented lens 14 mounted on the mounting base 110, and the double cemented lens 14 and the collimating lens 13 are disposed along the same optical axis direction and located downstream of the collimating lens 13. The collimated light passing through the collimating lens 13 is incident on the front surface of the double cemented lens 1414, and there is a better illumination spot at the focal plane of the double cemented lens 14. The center point of the collimator lens 13 and the center point of the first monochromatic light source 111 are on the same optical axis. The divergence angle of the first monochromatic light source 111 can be selected to be about 10 degrees, in order to enable as many light beams as possible to enter the collimating lens 13 and ensure that the processing space is enough, the linear distance between the two is 50mm, and the double cemented lens 14 is arranged on the 10mm position of the rear surface of the collimating lens 13 to play roles of achromatization and convergence.

Further, in one embodiment, the excitation light source module 10 further includes a fly-eye lens 15 mounted on the mounting base 110, the fly-eye lens 15 and the collimating lens 13 are disposed along the same optical axis direction, and the fly-eye lens 15 is located between the collimating lens 13 and the double-cemented lens 14. The light rays are changed into parallel light rays after passing through the collimating lens 13, pass through the fly-eye lens 15 and then strike the front surface of the double-cemented lens 1414, so that the size consistency and uniformity of different monochromatic light sources on the surface of the chip 70 are improved.

Referring to fig. 3 and 4, in one embodiment, the filter assembly 230 includes a fixing base 232, and an excitation filter 234, a fourth dichroic mirror 236 and an emission filter 238 mounted on the fixing base 232. Excitation filter 234 and emission filter 238 are perpendicular to each other, and fourth dichroic mirror 236 is 45 ° to excitation filter 234 and emission filter 238, respectively. The side of the fixing base 232 opposite to the emission filter 238 is provided with a fourth opening, when the turntable 220 rotates to a preset position, the excitation filter 234 is opposite to the first opening 201, the fourth opening is opposite to the second opening 202, and the emission filter 238 is opposite to the third opening. The light beams converged by the double-cemented lens 14 vertically enter the excitation filter 234 of one filter assembly 230 of the filter module 20, the excitation filter 234 is band-pass filter, only light beams in a target bandwidth pass through, the filtered light beams strike the surface of a fourth dichroic mirror 236 at an angle of 45 degrees with the light beams, the fourth dichroic mirror 236 changes the direction of the optical axis, so that the light beams horizontally propagating become vertically propagating, the light beams in the vertical direction vertically irradiate the horizontally placed chip 70 through the fourth opening and the second opening 202, the chip 70 is located on the focal plane of the double-cemented lens 14, droplets of the chip 70 emit fluorescent signals with a wavelength slightly larger than the wavelength of the excitation light due to stokes shift under the irradiation of the excitation light, the fluorescent signals in the vertical direction penetrate through the fourth dichroic mirror 236, the excitation light and the background light are filtered by the emission filter 238, and only the light beams in the target bandwidth enter the imaging module 40.

Further, in one embodiment, the base 210 includes a bottom shell 212 and an upper cover 214 connected to the bottom shell 212, the bottom shell 212 and the upper cover 214 enclose a receiving cavity for receiving the turntable 220 and the filter assembly 230, and the turntable 220 is rotatably connected to the bottom shell 212 and/or the upper cover 214. The first opening 201 is opened on the side of the bottom case 212, the third opening is opened on the ground of the bottom case 212, and the second opening 202 is opened on the upper cover 214. The filter assembly 230 is disposed in the accommodating cavity to prevent other light from affecting the detection.

Further, in one embodiment, the base 210 further includes a cover 216, the upper cover 214 defines an operation window 218 communicating with the accommodating cavity, and the cover 216 is rotatably connected to the upper cover 214 to open or close the operation window 218. The operating window 218 is exposed by rotating the cover 216, the filter assembly 230 located in the accommodating cavity is shifted through the operating window 218, so that the required filter assembly 230 rotates to the corresponding positions of the first opening 201, the second opening 202 and the third opening, and the cover 216 is rotated again after the filter assembly is rotated in place to seal the operating window 218, thereby preventing other light rays from entering the accommodating cavity.

Further, in one embodiment, the upper cover 214 is provided with a locking bead 219, and the locking bead 219 partially protrudes into the accommodating cavity, so that the fixing seat 232 is limited. Referring to fig. 3, the ball 219 is located on one side of the fixing seat 232 close to the center of the turntable 220, the fixing seat 232 can be slightly limited, the fixing seat 232 can overcome the limitation of the ball 219 under the action of external force and rotate along with the turntable 220, and the ball 219 is limited to the current position after rotating to the position.

Optionally, referring to fig. 1, in one embodiment, the carrier module 30 includes a first support frame 310, a first sliding bar 320, a second support frame 330, a second sliding bar 340, and a chip rack 350. The first support frame 310 is fixed to the base 210, and the first sliding rod 320 is fixed to the first support frame 310 and located above the base 210. The two support frames are slidably connected to the first sliding rod 320, and the second sliding rod 340 is fixed to the second support frame 330. The chip holder 350 is slidably connected to the second sliding bar 340, and the first sliding bar 320 is perpendicular to the second sliding bar 340. The chip frame 350 can slide along the second sliding rod 340, the second supporting frame 330 and the second sliding rod 340 can slide along the first sliding rod 320, and the carrier module 30 enables the chip 70 to be horizontally placed, move left and right and move front and back to change the position of the chip 70, so that switching of different imaging view fields is realized. In this embodiment, the first sliding rod 320 is fixed to the first support frame 310 and located above the base 210, so as to adjust the position of the chip 70.

In one embodiment, the imaging module 40 includes a lens 410 and an image sensor 420 connected to the lens 410, and the lens 410 is fixed to the lower end of the base 210. The imaging module 40 can be COMS, the cost is low, and the overall size of the device is designed as follows: 25.7cm 17.8cm 28.6cm, small in size and portable. The imaging module 40 is connected to the lower end of the base 210 by a connection member 430.

Specifically, the axis of the imaging module 40 is orthogonal to the optical axis of the excitation light source module 10, the axis of the imaging module 40 is also perpendicular to the surface of the chip 70, and the working distance of the lens 410 is 60mm, that is, the distance from the front surface of the lens 410 to the surface of the chip 70 is 60 mm. The plurality of filter assemblies 230 in the filter module 20 are distributed on the turntable 220 in a regular hexagon shape, and the turntable 220 can be rotated to switch different filter assemblies 230 to be in butt joint with the incident light in the horizontal direction and the lens 410 in the vertical direction, and the filter assemblies 230 can be slightly fixed by fixing a clamping ball after channels are selected.

In one embodiment, the digital PCR fluorescence detection apparatus further includes a supporting rod 50 and a fixing plate 60, wherein one end of the supporting rod 50 is connected to the base 210, the other end of the supporting rod 50 is connected to the fixing plate 60, and the length of the supporting rod 50 is greater than the length of the imaging module 40. The whole device is placed at a certain position through the fixing plate 60, and a certain operation space is formed between the base 210 and the fixing plate 60 through the supporting rod 50.

Compared with the traditional flow cytometry detection method, the method has the advantages that one microdrop passes through the detection area to detect the intensity of the fluorescence signal; conventional imaging CCD imaging is limited by the imaging field of view, and neither method is flux-high. The digital PCR fluorescence detection device of the application combines the excitation light source of the beam by a plurality of monochromatic light sources; a multi-channel filtering module 20 is arranged, and a filtering component of a channel is switched by rotating; the imaging device uses CMOS; the volume is small, the cost is reduced, the utilization rate of light energy is improved, and the detection flux and the detection efficiency are improved.

In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the invention.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

In the present invention, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may be directly contacting the second feature or the first and second features may be indirectly contacting each other through intervening media. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.

It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like as used herein are for illustrative purposes only and do not denote a unique embodiment.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims (10)

1. A digital PCR fluorescence detection device is characterized by comprising

The excitation light source module comprises a mounting seat, a dichroic mirror and a plurality of monochromatic light sources, wherein an exit port is formed in one end of the mounting seat, the monochromatic light sources are arranged at the other end of the mounting seat in parallel or vertically, the dichroic mirror and the monochromatic light sources are arranged at an angle of 45 degrees, and light rays of the monochromatic light sources are transmitted or reflected to the exit port to be emitted through the dichroic mirror;

the filtering module comprises a base and a turntable which is rotationally connected with the base, the turntable is provided with a plurality of filtering components along the circumferential direction of the turntable, the base is provided with a first opening, a second opening and a third opening, and the first opening corresponds to the emergent port;

the carrying module is arranged on one side of the base and used for carrying a chip to be detected, and the second opening faces the carrying module;

and the imaging module is arranged on the other side of the base, and the third opening faces the imaging module.

2. The digital PCR fluorescence detection device of claim 1, further comprising a collimating lens disposed on the mounting base, wherein the plurality of monochromatic light sources includes a first monochromatic light source and a second monochromatic light source, the dichroic mirror includes a first dichroic mirror and a second dichroic mirror, the first monochromatic light source and the second monochromatic light source are perpendicular to each other and are axially symmetric with respect to the first dichroic mirror, the first dichroic mirror and the second dichroic mirror are perpendicular to each other, and the first dichroic mirror, the second dichroic mirror and the collimating lens are sequentially disposed along an optical axis direction of the first monochromatic light source.

3. The digital PCR fluorescence detection device of claim 2, wherein the excitation light source module further comprises a third monochromatic light source, a fourth monochromatic light source and a third dichroic mirror mounted on the mounting base, the third monochromatic light source and the fourth monochromatic light source are perpendicular to each other and are axially symmetric with respect to the third dichroic mirror, the third dichroic mirror and the second dichroic mirror are parallel to each other, an extension line of the first monochromatic light source is perpendicular to an extension line of the fourth monochromatic light source, the second monochromatic light source is perpendicular to an extension line of the third monochromatic light source, distances from a midpoint of the first monochromatic light source and the fourth monochromatic light source to the second dichroic mirror are equal, and the first monochromatic light source and the fourth monochromatic light source are axially symmetric with respect to the second dichroic mirror.

4. The digital PCR fluorescence detection device of claim 3, wherein the excitation light source module further includes a double cemented lens mounted on the mounting base, and the double cemented lens and the collimating lens are disposed along a same optical axis direction and located downstream of the collimating lens.

5. The digital PCR fluorescence detection device of claim 4, wherein the excitation light source module further includes a fly-eye lens mounted on the mounting base, the fly-eye lens and the collimating lens are disposed along a same optical axis direction, and the fly-eye lens is located between the collimating lens and the double-cemented lens.

6. The digital PCR fluorescence detection device according to any one of claims 1-5, wherein the filter assembly comprises a fixed seat, and an excitation filter, a fourth dichroic mirror and an emission filter which are arranged on the fixed seat, the excitation filter and the emission filter are perpendicular to each other, the fourth dichroic mirror and the excitation filter and the emission filter are 45 degrees respectively, a fourth opening is arranged on the side surface of the fixed seat opposite to the emission filter, when the rotating disk rotates to a preset position, the excitation filter is opposite to the first opening, the fourth opening is opposite to the second opening, and the emission filter is opposite to the third opening.

7. The digital PCR fluorescence detection device of claim 6, wherein the base comprises a bottom shell and an upper cover connected with the bottom shell, the bottom shell and the upper cover enclose a cavity for accommodating the turntable and the filter assembly, and the turntable is rotatably connected with the bottom shell and/or the upper cover.

8. The digital PCR fluorescence detection device of claim 7, wherein the base further comprises a cover, the cover is provided with an operation window communicated with the accommodating cavity, and the cover is rotatably connected with the cover to open or close the operation window;

and/or the upper cover is provided with a clamping bead, and the clamping bead partially protrudes into the accommodating cavity so that the fixing seat is limited.

9. The digital PCR fluorescence detection device of claims 1-5, wherein the carrier module comprises a first support frame, a first slide bar, a second support frame, a second slide bar and a chip holder, the first support frame is fixed on the base, the first slide bar is fixed on the first support frame and positioned above the base, the second support frame is slidably connected with the first slide bar, the second slide bar is fixed on the second support frame, the chip holder is slidably connected with the second slide bar, and the first slide bar and the second slide bar are perpendicular to each other;

and/or, the imaging module comprises a lens and an image sensor connected with the lens, and the lens is fixed at the lower end of the base.

10. The digital PCR fluorescence detection device according to any one of claims 1-5, further comprising a support rod and a fixing plate, wherein one end of the support rod is connected to the base, the other end of the support rod is connected to the fixing plate, and the length of the support rod is greater than the length of the imaging module.

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