Optical detection device for polymerase chain reaction

Abstract

In the embodiment of the invention, the optical detection device for the polymerase chain reaction comprises a micro-droplet detection flow channel for micro-droplets to be detected to flow through; a first excitation light module and a second excitation light module, wherein each excitation light module comprises an excitation light emitter and an excitation light filtering piece; a first fluorescence detection module and a second fluorescence detection module, wherein each fluorescence detection module comprises a fluorescence detector and a fluorescence filtering piece; wherein the maximum wavelength of the passband in the first exciting light filtering piece and the second exciting light filtering piece is smaller than the minimum wavelength of the passband in the first fluorescent light filtering piece and the second fluorescent light filtering piece. Therefore, in the application, the passband ranges of the filter elements are arranged in the spectrum according to the sequence of the exciting light filter element 1, the exciting light filter element 2,..., the fluorescent light filter element 1, the fluorescent light filter element 2,..., so that the passband ranges of the filter elements are wider, and the excitation efficiency and the fluorescent light receiving efficiency of the dye are improved.

Description

Technical Field

[0001] This application relates to the field of optical detection technology, and in particular to an optical detection device for polymerase chain reaction. Background Art

[0002] Fluorescence is a physical phenomenon of light emission. That is, under the action of excitation light with a first wavelength, due to the Stokes shift phenomenon, emitted light with a longer second wavelength is generated, namely fluorescence. Since the energy of the excitation light is much greater than the energy of the fluorescence, even if a small part of the excitation light is detected by the detector, it will cause the detector to saturate or reduce the measurement range of the detection system. In the optical detection system of qPCR (Quantitative Real-time PCR) technology, a common solution to avoid excitation light from entering the detector is to arrange an excitation light filter and an emission light filter at the peak positions of the excitation light wavelength and the emission light wavelength corresponding to the fluorescent dye, respectively. When there are many detection channels, in order to avoid the overlap range between the emission light band and the excitation light band of different fluorescent dyes, usually more than 4 filters need to be arranged within a 100nm bandwidth range.

[0003] However, the problem with the above solution is that the passband of the filter is usually narrow. The narrow passband range of the filter will reduce the excitation efficiency of the excitation light and the detection efficiency of the emission light. If it is applied to the fluorescence signal detection of microdroplets with a small size, the signal-to-noise ratio will be very low. Summary of the Invention

[0004] The embodiment of this application provides an optical detection device for polymerase chain reaction, which reselects and combines the filter components of the optical solution. The passband ranges of the filter components are arranged in the spectrum in the order of excitation light filter component 1, excitation light filter component 2,... fluorescence filter component 1, fluorescence filter component 2,... so that the passband ranges of the filter components are wider, which is beneficial to improving the excitation efficiency of the fluorescent dye and the reception efficiency of the fluorescence.

[0005] An optical detection device for polymerase chain reaction provided by an embodiment of the present application includes: a droplet detection flow channel for allowing a fluid to be measured to flow through, where the fluid to be measured at least includes droplets to be detected; a first excitation light module including a first excitation light emitter and a first excitation light filter; a second excitation light module including a second excitation light emitter and a second excitation light filter; a first fluorescence detection module including a first fluorescence detector and a first fluorescence filter; a second fluorescence detection module including a second fluorescence detector and a second fluorescence filter; the first excitation light emitter and the second excitation light emitter respectively emit a first excitation light and a second excitation light, and after the first excitation light and the second excitation light are filtered by the first excitation light filter and the second excitation light filter respectively, they converge and irradiate into the droplet detection flow channel, so that at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence correspondingly, and one path of the fluorescence is received by the first fluorescence detector after being filtered by the first fluorescence filter, and the other path of the fluorescence is received by the second fluorescence detector after being filtered by the second fluorescence filter; wherein, the maximum wavelength of the passband in the first excitation light filter and the second excitation light filter is less than the minimum wavelength of the passband in the first fluorescence filter and the second fluorescence filter.

[0006] In some embodiments, the passbands of the first excitation light filter and the second excitation light filter are separated from each other; or the passbands of the first excitation light filter and the second excitation light filter partially overlap.

[0007] In some embodiments, the passbands of the first fluorescence filter and the second fluorescence filter are separated from each other; or the passbands of the first fluorescence filter and the second fluorescence filter partially overlap.

[0008] In some embodiments, it further includes a third excitation light module and the third fluorescence detection module; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; the third excitation light emitter emits a third excitation light, and after the third excitation light passes through the third excitation light filter, it converges with the first excitation light and the second excitation light and irradiates into the droplet detection flow channel, so that at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence correspondingly, and one path of the fluorescence is received by the third fluorescence detector after being filtered by the third fluorescence filter; wherein, the maximum wavelength of the passband in the first excitation light filter, the second excitation light filter and the third excitation light filter is less than the minimum wavelength of the passband in the first fluorescence filter, the second fluorescence filter and the third fluorescence filter.

[0009] In some embodiments, a third excitation light module and the third fluorescence detection module are further included; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; the third excitation light emitter emits third excitation light, and after the third excitation light passes through the third excitation light filter, it converges with the first excitation light and the second excitation light and irradiates into the droplet detection flow channel, so that at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence correspondingly, and one path of the fluorescence is received by the third fluorescence detector after being filtered by the third fluorescence filter; wherein, the maximum wavelength of the passband of the third excitation light filter is less than the minimum wavelength of the passband of the third fluorescence filter, and the minimum wavelength of the passband of the third excitation light filter is greater than the maximum wavelength of the passbands of the first fluorescence filter and the second fluorescence filter.

[0010] In some embodiments, a third excitation light module, the third fluorescence detection module, a fourth excitation light module and the fourth fluorescence detection module are further included; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; the fourth excitation light module includes a fourth excitation light emitter and a fourth excitation light filter; the fourth fluorescence detection module includes a fourth fluorescence detector and a fourth fluorescence filter; the third excitation light emitter and the fourth excitation light emitter respectively emit third excitation light and fourth excitation light, and after the third excitation light and the fourth excitation light respectively pass through the third excitation light filter and the fourth excitation light filter, the first excitation light, the second excitation light, the third excitation light and the fourth excitation light converge and irradiate into the droplet detection flow channel, so that at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence correspondingly, one path of the fluorescence is received by the third fluorescence detector after being filtered by the third fluorescence filter, and one path of the fluorescence is received by the fourth fluorescence detector after being filtered by the fourth fluorescence filter; wherein, the minimum wavelength of the passbands of the third excitation light filter and the fourth excitation light filter is greater than the maximum wavelength of the passbands of the first fluorescence filter and the second fluorescence filter, and the maximum wavelength of the passbands of the third excitation light filter and the fourth excitation light filter is less than the minimum wavelength of the passbands of the third fluorescence filter and the fourth fluorescence filter.

[0011] In some embodiments, the passband of the third excitation light filter and the passband of the fourth excitation light filter are separated from each other; or the passband of the third excitation light filter and the passband of the fourth excitation light filter partially overlap.

[0012] In some embodiments, the passband of the third fluorescence filter and the passband of the fourth fluorescence filter are separated from each other; or the passband of the third fluorescence filter and the passband of the fourth fluorescence filter partially overlap.

[0013] In some embodiments, a first filter is further included; the droplet to be detected emits fluorescence, and after being filtered by the first filter, one path of the fluorescence is received by the first fluorescence detector after being filtered by the first fluorescence filter, and the other path of the fluorescence is received by the second fluorescence detector after being filtered by the second fluorescence filter, wherein the maximum wavelength of the passband in the first excitation light filter and the second excitation light filter is less than the minimum wavelength of the passband of the first filter; and / or a first dichroic mirror is further included, the first fluorescence detector and the first fluorescence filter are both disposed on the transmission optical path of the first dichroic mirror, and the second fluorescence detector and the second fluorescence filter are both disposed on the reflection optical path of the first dichroic mirror, wherein the cut-off wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband in the first fluorescence filter and the second fluorescence filter, and less than the maximum wavelength of the passband in the first fluorescence filter and the second fluorescence filter.

[0014] In some embodiments, a second filter is further included; the droplet to be detected correspondingly emits fluorescence, and after being filtered by the second filter, one path of the fluorescence is received by the third fluorescence detector after being filtered by the third fluorescence filter, and the other path of the fluorescence is received by the fourth fluorescence detector after being filtered by the fourth fluorescence filter, wherein the maximum wavelength of the passband in the third excitation light filter and the fourth excitation light filter is less than the minimum wavelength of the passband of the second filter; and / or a second dichroic mirror is further included, the third fluorescence detector and the third fluorescence filter are both disposed on the transmission optical path of the second dichroic mirror, and the fourth fluorescence detector and the fourth fluorescence filter are both disposed on the reflection optical path of the second dichroic mirror; wherein the cut-off wavelength of the second dichroic mirror is greater than the minimum wavelength of the passband in the third fluorescence filter and the fourth fluorescence filter, and less than the maximum wavelength of the passband in the third fluorescence filter and the fourth fluorescence filter.

[0015] In an embodiment of the present application, an optical detection device for polymerase chain reaction includes: a droplet detection flow channel for allowing a fluid to be measured to flow through, where the fluid to be measured at least includes droplets to be detected; a first excitation light module including a first excitation light emitter and a first excitation light filter; a second excitation light module including a second excitation light emitter and a second excitation light filter; a first fluorescence detection module including a first fluorescence detector and a first fluorescence filter; a second fluorescence detection module including a second fluorescence detector and a second fluorescence filter; the first excitation light emitter and the second excitation light emitter respectively emit a first excitation light and a second excitation light, and after the first excitation light and the second excitation light are respectively filtered by the first excitation light filter and the second excitation light filter, they are converged and irradiated into the droplet detection flow channel, so that at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence accordingly. One path of fluorescence is received by the first fluorescence detector after being filtered by the first fluorescence filter, and the other path of fluorescence is received by the second fluorescence detector after being filtered by the second fluorescence filter; wherein, the maximum wavelength of the passband in the first excitation light filter and the second excitation light filter is less than the minimum wavelength of the passband in the first fluorescence filter and the second fluorescence filter. Therefore, in the present application, the passband ranges of the filters are arranged in the spectrum in the order of excitation light filter 1, excitation light filter 2,..., fluorescence filter 1, fluorescence filter 2,..., so that the passband ranges of the filters are wider, and further, the excitation wavelength range of the fluorescent dye is larger, and at the same time, the fluorescence wavelength range of the fluorescent dye is larger, which is beneficial to improving the excitation efficiency of the fluorescent dye and the reception efficiency of fluorescence at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 FIG. is a schematic diagram of the wavelength ranges of the excitation light and fluorescence filters used in the prior art provided by an embodiment of the present application;

[0017] Figure 2 FIG. is a structural block diagram of an optical detection device provided by an embodiment of the present application;

[0018] Figure 3 FIG. is a structural schematic diagram of the first and second fluorescence detection modules provided by an embodiment of the present application;

[0019] Figure 4 FIG. is a structural schematic diagram of the first and second excitation light modules provided by an embodiment of the present application;

[0020] Figure 5 FIG. is a schematic diagram of the excitation spectra of the first and second fluorescent dyes provided by an embodiment of the present application;

[0021] Figure 6 FIG. is a schematic diagram when the passbands of the first and second excitation light filters and the first and second fluorescence filters overlap provided by an embodiment of the present application;

[0022] Figure 7 Schematic diagram when the passbands of the first and second excitation light filters and the first and second fluorescence filters provided by the embodiments of the present application are separated;

[0023] Figure 8 Schematic diagram of the emission spectra of the first and second fluorescent dyes provided by the embodiments of the present application;

[0024] Figure 9 Schematic diagram of the structures of the first, second, and third fluorescence detection modules provided by the embodiments of the present application;

[0025] Figure 10 Schematic diagram of the structures of the first, second, and third excitation light modules provided by the embodiments of the present application;

[0026] Figure 11 Schematic diagram when the passbands of the three excitation light filters and the three fluorescence filters provided by the embodiments of the present application overlap;

[0027] Figure 12 Schematic diagram when the passbands of the three excitation light filters and the three fluorescence filters provided by the embodiments of the present application are separated;

[0028] Figure 13 Another schematic diagram when the passbands of the three excitation light filters and the three fluorescence filters provided by the embodiments of the present application overlap;

[0029] Figure 14 Schematic diagram of the structure of a first, second, third, and fourth fluorescence detection module provided by the embodiments of the present application;

[0030] Figure 15 Schematic diagram of the structures of the first, second, third, and fourth excitation light modules provided by the embodiments of the present application. Detailed implementation manners

[0031] In order to enable those skilled in the art to better understand the solution of the present application, the technical solutions in the present application will be clearly and completely described below with reference to the accompanying drawings in the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present application.

[0032] Fluorescence is a physical phenomenon of light emission, that is, under the action of excitation light of a first wavelength, due to the Stokes shift phenomenon, emission light of a second wavelength that is longer in wavelength is generated, that is, fluorescence. As shown in the excitation spectrum and emission spectrum of Dye 1 in Figure 1 . Different fluorescent dyes have different excitation light wavelengths and emission light wavelengths, and have different excitation efficiencies and emission efficiencies at different wavelengths.

[0033] Since the energy of the excitation light is much greater than that of the fluorescence, even if a small part of the excitation light is detected by the detector, it will cause the detector to saturate or reduce the measurement range of the detection system. In the optical detection system of qPCR (Quantitative Real-time PCR) technology, a common solution to avoid excitation light from entering the detector is to arrange an excitation light filter and an emission light filter at the peak positions of the excitation light wavelength and the emission light wavelength of the corresponding fluorescent dye, respectively. When there are many detection channels, the excitation / emission spectra of the fluorescent dyes are arranged in the order of fluorescence 1 excitation, fluorescence 1 emission, fluorescence 2 excitation, fluorescence 2 emission, fluorescence 3 excitation, fluorescence 3 emission,... See Figure 1 , Figure 1 FIG. 4 is a schematic diagram of the excitation spectra, emission spectra of four fluorescent dyes, and the passband ranges of the excitation light filter element and the fluorescence filter element. The colored rectangular areas represent the passband ranges of the filters, that is, from left to right are the passband range of the first excitation light filter element, the passband range of the first fluorescence filter element, the passband range of the second excitation light filter element, the passband range of the second fluorescence filter element, the passband range of the third excitation light filter element, the passband range of the third fluorescence filter element, the passband range of the fourth excitation light filter element, and the passband range of the fourth fluorescence filter element (shown as excitation 1, fluorescence 1, excitation 2, fluorescence 2, excitation 3, fluorescence 3, excitation 4, fluorescence 4 in the figure); the dotted line is the excitation spectrum, and the line segment is the emission spectrum, that is, from left to right are the excitation light wavelength of the first dye, the emission light wavelength of the first dye, the excitation light wavelength of the second dye, the emission light wavelength of the second dye, the excitation light wavelength of the third dye, the emission light wavelength of the third dye, the excitation light wavelength of the fourth dye, and the emission light wavelength of the fourth dye. The overlapping parts of the rectangular areas with the excitation spectra and emission spectra of the corresponding dyes represent the fluorescence excitation efficiency and fluorescence reception efficiency. The larger the overlapping area, the higher the excitation and detection efficiency. However, the problem with the above solution is that the passband of the filter is usually narrow, resulting in low excitation efficiency and emission efficiency. If it is applied to the fluorescence signal detection of microdroplets with a small size, the signal-to-noise ratio will be very low, which may affect the detection result.

[0034] The embodiment of the present application provides an optical detection device 1 for polymerase chain reaction, which reselects and combines the filter elements of the optical solution. The passband ranges of the filter elements are arranged in the spectrum in the order of excitation light filter element 1, excitation light filter element 2,... fluorescence filter element 1, fluorescence filter element 2,... so that the passband ranges of the filter elements are wider, thereby making the excitation light wavelength range of the fluorescent dye larger, improving the excitation efficiency of the dye, and making the fluorescence wavelength range of the fluorescent dye larger, improving the fluorescence reception efficiency. For the optical detection device 1 for polymerase chain reaction provided by the present application, please refer to the following content and Figures 2 - 15 :

[0035] See Figure 2 The optical detection device 1 includes a droplet detection flow channel 10, at least two excitation light modules 20, and at least two fluorescence detection modules 30. The droplet detection flow channel 10 is used for the fluid to be detected to flow through. The fluid to be detected at least includes droplets 11 to be detected, and at least two fluorescent dyes are included in the droplets 11 to be detected. The excitation light module 20 is used to generate excitation light and excite the fluorescent dyes in the droplets 11 to be detected to generate fluorescence. The fluorescence detection module 30 is used to receive the fluorescence to obtain the detection data of the droplets 11 to be detected.

[0036] Among them, the numbers of the excitation light module 20 and the fluorescence detection module 30 are not limited. Preferably, the numbers of the excitation light module 20 and the fluorescence detection module 30 correspond one by one. For example, the optical detection device 1 may include two excitation light modules 20 and two fluorescence detection modules 30, or the optical detection device 1 may include four excitation light modules 20 and four fluorescence detection modules 30. Of course, it may also include any other numbers of excitation light modules 20 and fluorescence detection modules 30.

[0037] In some embodiments, the excitation light module 20 includes an excitation light emitter and an excitation light filter. Among them, the excitation light emitter is used to generate excitation light, and the excitation light can excite a fluorescent substance to generate fluorescence. The excitation light can be generated by a laser, an LED, a halogen lamp, or other light sources. Different excitation light emitters are used to generate excitation light in different wavelength ranges, or the excitation light emitter can generate excitation light in different wavelength ranges after being modulated. Among them, the filter is used to screen light in a specific wavelength range, and can filter out unnecessary bands and only retain light in a specific wavelength range to pass through.

[0038] The filter in the embodiments of the present application is a filter sheet, that is, the filter is a sheet-like component. In some embodiments, the filter may also be a filter lens.

[0039] In the embodiments of the present application, the excitation light emitted by the excitation light emitter is irradiated on the droplet detection flow channel 10 after being filtered by the excitation light filter.

[0040] In some embodiments, the optical detection device 1 further includes a spot shaping device 401, see Figure 4 , which is used to converge and shape the excitation light passing through the excitation light filter to form a spot and then irradiate it on the droplet detection flow channel 10.

[0041] In some embodiments, the excitation light emitted by at least two excitation light modules 20 can be converged onto the same optical path through a dichroic mirror 402 and then emitted to the spot shaping device 401, for example, see Figure 4The excitation light optical path shown in [figure reference], or the excitation light emitted by a single excitation light module 20 can independently use an optical path to be emitted to the spot shaping device 401. It should be noted that the above two methods can be used alone or in combination in the setting mode of at least two excitation light modules 20, and the present application does not make specific restrictions on this.

[0042] In some embodiments, at least one excitation light module 20 further includes a brightness feedback module 403. For example Figure 4 the brightness feedback module 403 shown in [figure reference], the brightness feedback module 403 includes a brightness detector. A part of the excitation light generated by the excitation light emitter is transmitted through the dichroic mirror of the brightness feedback module 403 and detected by the brightness detector. The brightness detector feeds back the detected excitation light brightness to the processor of the optical detection device 1, and the processor then closed-loop controls the brightness of the excitation light generated by the excitation light emitter. In some embodiments, the brightness feedback module 403 further includes a filter and a focusing lens.

[0043] In some embodiments, the fluorescence detection module 30 includes a fluorescence detector and a fluorescence filter. The droplet to be detected 11 flowing through the droplet detection channel 10 emits fluorescence under the irradiation of the excitation light, and the emitted fluorescence is filtered by the fluorescence filter and then received by the fluorescence detector.

[0044] Among them, each of at least two fluorescence detection modules 30 can be flexibly arranged on any optical path of the droplet detection channel 10. Specifically, each of at least two fluorescence detection modules 30 can be independently arranged on an optical path of the droplet detection channel 10; or two or more fluorescence detection modules 30 can be combined and arranged on an optical path of the droplet detection channel 10, for example, see Figure 3 . The above two methods can be used independently or in combination, and the present application does not make specific restrictions on this.

[0045] In some embodiments, when two or more fluorescence detection modules 30 are combined and arranged on the same optical path of the droplet detection channel 10, a dichroic mirror is further included between every two fluorescence detection modules 30. Exemplarily, see Figure 3 , a first dichroic mirror 51 is further arranged between the first fluorescence detection module 31 and the second fluorescence detection module. Among them, the dichroic mirror has a cut-off wavelength, including a long-pass dichroic mirror and a short-pass dichroic mirror. The long-pass dichroic mirror is used to pass light rays greater than its cut-off wavelength, and to reflect light rays less than its cut-off wavelength. The short-pass dichroic mirror is used to pass light rays less than its cut-off wavelength, and to reflect light rays less than its cut-off wavelength. Among them, the optical path through which the light rays pass hereinafter is called the transmission optical path, and the optical path for reflecting the light rays is called the reflection optical path.

[0046] In some embodiments, on the optical path of the fluorescence detection module 30 provided in the droplet detection channel 10, between the droplet detection channel 10 and the fluorescence detection module 30, a filter is further provided to filter the excitation light and prevent the excitation light from entering the fluorescence detection module 30. Thus, under the double filtering of this filter and the fluorescence filter, the detection result is more accurate and effective.

[0047] In the embodiments of the present application, the optical detection device 1 includes at least two excitation light modules 20 and at least two fluorescence detection modules 30. The arrangement order of the passband ranges of at least two excitation light filters and at least two fluorescence filters can be determined according to relevant factors such as the excitation efficiency and reception efficiency of each fluorescent dye, and by selecting the filter passbands with the highest excitation efficiency and reception efficiency.

[0048] The following uses specific embodiments to illustrate the arrangement modes of the passbands of the excitation light filters and the fluorescence filters when different numbers of excitation light modules 20 and fluorescence detection modules 30 are included.

[0049] In some embodiments, the optical detection device 1 includes two excitation light modules 20 and two fluorescence detection modules 30. Specifically, see Figures 3 - 8 :

[0050] The optical detection device 1 includes: a droplet detection channel 10, a first excitation light module 21, a second excitation light module 22, a first fluorescence detection module 31, and a second fluorescence detection module 32. Among them, the first excitation light module 21 includes a first excitation light emitter 211 and a first excitation light filter 212; the second excitation light module 22 includes a second excitation light emitter 221 and a second excitation light filter 222; the first fluorescence detection module 31 includes a first fluorescence detector 311 and a first fluorescence filter 312; the second fluorescence detection module 32 includes a second fluorescence detector 321 and a second fluorescence filter 322. The first excitation light emitter 211 and the second excitation light emitter 221 respectively emit a first excitation light and a second excitation light. After the first excitation light and the second excitation light are respectively filtered by the first excitation light filter 212 and the second excitation light filter 222, they are converged and irradiated into the droplet detection channel 10, so that at least some of the droplets 11 to be detected flowing through the droplet detection channel 10 emit fluorescence accordingly. One path of fluorescence is received by the first fluorescence detector 311 after being filtered by the first fluorescence filter 312, and the other path of fluorescence is received by the second fluorescence detector 321 after being filtered by the second fluorescence filter 322. At least some of the droplets 11 to be detected here contain fluorescent dyes that can be excited by the excitation light passing through the first excitation light filter 212 and / or the second excitation light filter 222.

[0051] Exemplarily, see Figures 3 - 4 , Figure 3Schematic diagram of a structure provided by an embodiment of the present application, which includes two fluorescence detection modules 30. Figure 4 Schematic diagram of a structure provided by an embodiment of the present application, which includes two excitation light modules 20. As Figure 3 shown, the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the same optical path of the droplet detection flow channel 10. As Figure 4 shown, the emitted light from the first excitation light module 21 and the second excitation light module 22 is converged onto the same optical path through a dichroic mirror, and is shaped by a spot shaping device 401 to form a spot and then irradiate on the droplet detection flow channel 10. It should be noted that Figure 3 and Figure 4 are only examples of the optical detection device 1 provided by the present application. The first fluorescence detection module 31, the second fluorescence detection module 32, the first excitation light module 21, and the second excitation light module 22 can of course also be assembled in other arrangements, and the present application does not make specific restrictions on this. For example, in Figure 3 and Figure 4 , a lens 71 can be provided between the fluorescence filter and the fluorescence detector, between the excitation light emitter and the excitation light filter, and between the droplet detection flow channel and the fluorescence detection module 30 to converge the excitation light or fluorescence and then emit it to the subsequent device.

[0052] Among them, the arrangement order of the passband ranges of the filters is the first excitation light filter 212, the second excitation light filter 222, the first fluorescence filter 312, and the second fluorescence filter 322. Specifically, in some embodiments, the maximum passband wavelength in the first excitation light filter 212 and the second excitation light filter 222 is less than the minimum passband wavelength in the first fluorescence filter 312 and the second fluorescence filter 322. The maximum passband wavelength of the first excitation light filter 212 is less than or equal to the maximum passband wavelength of the second excitation light filter 222. The maximum passband wavelength of the first fluorescence filter 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter 322.

[0053] Among them, the passband ranges of the excitation light filter and the fluorescence filter should not have an overlapping part, and the passband range of the excitation light filter is less than the passband range of the fluorescence filter to prevent the excitation light from entering the fluorescence detector. Specifically, the difference between the two can be adjusted according to actual needs and detection effects, and generally can be set to 10 nm.

[0054] Among them, the first fluorescence detection module 31 and the second fluorescence detection module 32 can be arranged on the same optical path of the droplet detection flow channel 10, or can be arranged on different optical paths of the droplet detection flow channel 10.

[0055] Specifically, in some embodiments, the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the same optical path of the droplet detection flow channel 10. A first dichroic mirror 51 is also arranged between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the transmission optical path or the reflection optical path of the first dichroic mirror 51. The cut-off wavelength of the first dichroic mirror 51 is greater than the minimum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322, and less than the maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322. The first dichroic mirror 51 is mainly used to arrange the first fluorescence detection module 31 and the second fluorescence detection module 32 on the same optical path, which can save the internal space of the optical detection device 1. Exemplarily, see Figure 3 , the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the same side of the droplet detection flow channel 10. A first dichroic mirror 51 is included between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first dichroic mirror 51 is a long-pass dichroic mirror. The first fluorescence detection module 31 is arranged on the reflection optical path of the first dichroic mirror 51, and the second fluorescence detection module 32 is arranged on the transmission optical path of the first dichroic mirror 51. Of course, the first dichroic mirror 51 can also be a short-pass dichroic mirror, and the specific setting method will not be elaborated here.

[0056] Further, on the optical path where the fluorescence detection module 30 is arranged in the droplet detection flow channel 10, between the droplet detection flow channel 10 and the fluorescence detection module 30, a filter is also arranged. For example, between the droplet detection flow channel 10 and the first fluorescence detection module 31, or between the droplet detection flow channel 10 and the second fluorescence detection module 32, a first filter 61 is arranged. The minimum passband wavelength of the first filter 61 is greater than the maximum passband wavelength of the first excitation light filter 212 and the second excitation light filter 222 to filter the excitation light and prevent the excitation light from entering the fluorescence detection module. For example, see Figure 3 the first filter 61 in

[0057] In some embodiments, the passbands of the first excitation light filter 212 and the second excitation light filter 222 are separated from each other; or the passbands of the first excitation light filter 212 and the second excitation light filter 222 partially or completely overlap.

[0058] Specifically, see Figure 5 , Figure 5The excitation spectra of the first fluorescent dye and the second fluorescent dye are shown. There is an overlapping part between the excitation spectrum of the first fluorescent dye and the excitation spectrum of the second fluorescent dye. The overlapping wavelength range can excite both the first fluorescent dye and the second fluorescent dye. Within the wavelength range of this overlapping part, the passband range of the first excitation light filter 212 can be set, and the passband range of the second excitation light filter 222 can also be set. Therefore, the passband ranges of the first excitation light filter 212 and the second excitation light filter 222 can overlap.

[0059] Among them, the overlapping degree or range of the passband ranges of the first excitation light filter 212 and the second excitation light filter 222 can be set flexibly. The passband ranges of the first excitation light filter 212 and the second excitation light filter 222 can either completely overlap or partially overlap. In some embodiments, the passband ranges of the first excitation light filter 212 and the second excitation light filter 222 partially overlap. As Figure 6 shown in the passband range of the first excitation light filter and the passband range of the second excitation light filter, the first fluorescent dye is mainly excited by the excitation light passing through the first excitation light filter 212, and can also be excited by a part of the excitation light (overlapping wavelength part) passing through the second excitation light filter 222. The second fluorescent dye is mainly excited by the excitation light passing through the second excitation light filter 222, and can also be excited by a part of the laser light (overlapping wavelength part) passing through the first excitation light filter 212. In some other embodiments, the passband ranges of the first excitation light filter 212 and the second excitation light filter 222 completely overlap, and the first fluorescent dye and the second fluorescent dye can be simultaneously excited by the first excitation light passing through the first excitation light filter 212 and the second excitation light passing through the second excitation light filter 222. Therefore, by overlapping the passbands of the first excitation light filter 212 and the second excitation light filter 222 completely or partially, the passband range of the filter can be increased, and the fluorescent dye can be excited by more than one excitation light to generate fluorescence, increasing the fluorescence excitation efficiency.

[0060] In some embodiments, the passband ranges of the first excitation light filter 212 and the second excitation light filter 222 are separated from each other. As Figure 7 shown, the excitation light passing through the first excitation light filter 212 is mainly used to excite the first fluorescent dye to generate fluorescence; the excitation light passing through the second excitation light filter 222 is mainly used to excite the second fluorescent dye to generate fluorescence.

[0061] In some embodiments, the passbands of the first fluorescence filter 312 and the second fluorescence filter 322 are separated from each other; or at least part of the passbands of the first fluorescence filter 312 and the second fluorescence filter 322 partially overlap.

[0062] Specifically, see Figure 8 shown.Figure 8 are the emission spectra of the first fluorescent dye and the second fluorescent dye. There is an overlapping part between the emission spectrum of the first fluorescent dye and the emission spectrum of the second fluorescent dye. The fluorescence filter in this overlapping wavelength range can pass both the first fluorescence generated by the first fluorescent dye and the second fluorescence generated by the second fluorescent dye. In the wavelength range of this overlapping part, the passband range of the first fluorescence filter 312 can be set, and the passband range of the second fluorescence filter 322 can also be set. Therefore, the passband ranges of the first fluorescence filter 312 and the second fluorescence filter 322 can at least partially overlap.

[0063] Among them, the overlapping range of the first fluorescence filter 312 and the second fluorescence filter 322 can be flexibly set. In some embodiments, there is a partial overlap between the passband ranges of the first fluorescence filter 312 and the second fluorescence filter 322. See Figure 6 for the settings of the passband ranges of the first fluorescence filter and the second fluorescence filter in. Then, for the detection results of the first fluorescence detector 311 and the second fluorescence detector 321, accurate detection results of each fluorescence can be obtained through post - fluorescence calibration. In some other embodiments, the passband ranges of the first fluorescence filter 312 and the second fluorescence filter 322 can also completely overlap. Therefore, by overlapping all or part of the passbands of the first fluorescence filter 312 and the second fluorescence filter 322, the passband range of the filter can be increased, and the fluorescence reception efficiency is improved.

[0064] In some embodiments, the passband ranges of the first fluorescence filter 312 and the second fluorescence filter 322 are separated from each other. See Figure 7 for the settings of the passband ranges of the first fluorescence filter and the second fluorescence filter in.

[0065] In some embodiments, the optical detection device 1 includes three excitation light modules 20 and three fluorescence detection modules 30. Specifically, see Figures 9 - 13 :

[0066] The optical detection device 1 includes: a droplet detection flow channel 10, a first excitation light module 21, a second excitation light module 22, a third excitation light module 23, a first fluorescence detection module 31, a second fluorescence detection module 32, and a third fluorescence detection module 33. Among them, the first excitation light module 21 includes a first excitation light emitter 211 and a first excitation light filter 212; the second excitation light module 22 includes a second excitation light emitter 221 and a second excitation light filter 222; the third excitation light module 23 includes a third excitation light emitter 231 and a third excitation light filter 232; the first fluorescence detection module 31 includes a first fluorescence detector 311 and a first fluorescence filter 312; the second fluorescence detection module 32 includes a second fluorescence detector 321 and a second fluorescence filter 322; the third fluorescence detection module 33 includes a third fluorescence detector 331 and a third fluorescence filter 332. The first excitation light emitter 211, the second excitation light emitter 221, and the third excitation light emitter 231 respectively emit a first excitation light, a second excitation light, and a third excitation light. After the first excitation light, the second excitation light, and the third excitation light are respectively filtered by the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232, they are converged and irradiated onto the droplet detection flow channel 10, so that at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence. One path of fluorescence is received by the first fluorescence detector 311 after being filtered by the first fluorescence filter 312, one path of fluorescence is received by the second fluorescence detector 321 after being filtered by the second fluorescence filter 322, and another path of fluorescence is received by the third fluorescence detector 331 after being filtered by the third fluorescence filter 332. At least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light passing through the first excitation light filter 212, the second excitation light filter 222, and / or the third excitation light filter 232.

[0067] Exemplarily, see Figures 9 - 10 , Figure 9 is a schematic structural diagram of a structure including three fluorescence detection modules 30 provided by an embodiment of the present application. Figure 10 is a schematic structural diagram of a structure including three excitation light modules 20 provided by an embodiment of the present application. As Figure 9 shown, the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the first optical path of the droplet detection flow channel 10, and the third fluorescence detection module 33 is arranged on the second optical path of the droplet detection flow channel 10. The first optical path is different from the second optical path. As Figure 10 shown, the emitted light emitted by the first excitation light module 21, the second excitation light module 22, and the third excitation light module 23 is converged onto the same optical path through a dichroic mirror, and is shaped by a spot shaping device 401 to form a spot and then irradiated onto the droplet detection flow channel 10. It should be noted that Figure 9 and Figure 10This is only an example of the optical detection device 1 provided for this application. The first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, the first excitation light module 21, the second excitation light module 22, and the third excitation light module 23 can of course also be assembled with other arrangements and other components. This application does not make specific restrictions on this.

[0068] Among them, the arrangement order of the passband ranges of the filter elements includes the following two types:

[0069] The first type: the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332.

[0070] The second type: the first excitation light filter element 212, the second excitation light filter element 222, the first fluorescence filter element 312, the second fluorescence filter element 322, the third excitation light filter element 232, and the third fluorescence filter element 332.

[0071] Specifically, in the first arrangement, the maximum passband wavelength in the first excitation light filter element 212, the second excitation light filter element 222, and the third excitation light filter element 232 is less than the minimum passband wavelength in the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332. Among them, the maximum passband wavelength of the first excitation light filter element 212 is less than or equal to the maximum passband wavelength of the second excitation light filter element 222, which is less than or equal to the maximum passband wavelength of the third excitation light filter element 232. The maximum passband wavelength of the first fluorescence filter element 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter element 322, which is less than or equal to the maximum passband wavelength of the third fluorescence filter element 332.

[0072] Among them, the first fluorescence detection module 31, the second fluorescence detection module 32, and the third fluorescence detection module 33 can be arranged on the same optical path of the droplet detection flow channel 10, or can be arranged on different optical paths of the droplet detection flow channel 10. When at least two of the first fluorescence detection module 31, the second fluorescence detection module 32, and the third fluorescence detection module 33 are arranged on the same optical path, a dichroic mirror is further included between every two fluorescence detection modules 30. For example, a first dichroic mirror 51 is further arranged between the first fluorescence detection module 31 and the second fluorescence detection module 32, and a second dichroic mirror is further arranged between the second fluorescence detection module 32 and the third fluorescence detection module 33. The cut-off wavelength of the first dichroic mirror 51 is greater than the minimum passband wavelength among the first fluorescence filter 312 and the second fluorescence filter 322, and less than the maximum passband wavelength among the first fluorescence filter 312 and the second fluorescence filter 322. The cut-off wavelength of the second dichroic mirror is greater than the minimum passband wavelength among the second fluorescence filter 322 and the third fluorescence filter 332, and less than the maximum passband wavelength among the second fluorescence filter 322 and the third fluorescence filter 332.

[0073] Furthermore, on the optical path of the droplet detection flow channel 10 where the fluorescence detection module 30 is arranged, a filter is further arranged between the droplet detection flow channel 10 and the fluorescence detection module 30. For example, a first filter 61 is arranged between the droplet detection flow channel 10 and the first fluorescence detection module 31, or a first filter 61 is arranged between the droplet detection flow channel 10 and the second fluorescence detection module 32, or a first filter 61 is arranged between the droplet detection flow channel 10 and the third fluorescence detection module 33. The minimum passband wavelength of the first filter 61 is greater than the maximum passband wavelength among the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232, and less than the minimum passband wavelength among the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332.

[0074] In some embodiments, the passbands of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 are separated from each other; or there is partial or complete overlap among the passbands of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232.

[0075] Among them, the overlapping range of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 can be flexibly set. For example, the passband ranges of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 can have all or partial overlap. In some embodiments, there is partial overlap in the passband ranges of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232, such as Figure 11As shown by the passband ranges of the first excitation light filter, the second excitation light filter, and the third excitation light filter, the first fluorescent dye is mainly excited by the excitation light passing through the first excitation light filter 212, and can also be excited by a part of the excitation light (the overlapping wavelength part) passing through the second excitation light filter 222. The second fluorescent dye is mainly excited by the excitation light passing through the second excitation light filter 222, and can also be excited by a part of the laser light (the overlapping wavelength part) passing through the first excitation light filter 212 and the third excitation light filter 232. The third fluorescent dye is mainly excited by the excitation light passing through the third excitation light filter 232, and can also be excited by a part of the excitation light (the overlapping wavelength part) passing through the second excitation light filter 222. In some other embodiments, the passband ranges of at least two of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 completely overlap. Therefore, in this embodiment, by overlapping all or part of the passbands of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232, the passband range of the filter can be increased, and the fluorescent dye can be excited by more than one kind of excitation light to generate fluorescence, so that the fluorescence excitation efficiency is increased.

[0076] In some other embodiments, the passband ranges of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 are separated from each other, as Figure 12 As shown by the passband range of the first excitation light filter, the passband range of the second excitation light filter, and the passband range of the third excitation light filter, the excitation light passing through the first excitation light filter 212 is used to excite the first fluorescent dye to generate fluorescence; the excitation light passing through the second excitation light filter 222 is used to excite the second fluorescent dye to generate fluorescence; the excitation light passing through the third excitation light filter 232 is used to excite the third fluorescent dye to generate fluorescence.

[0077] In some embodiments, the passbands of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332 are separated from each other. As Figure 12 As shown by the passband range of the first fluorescence filter, the passband range of the second fluorescence filter, and the passband range of the third fluorescence filter; or some or all of the passbands of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332 overlap, as Figure 11 As shown by the passband range of the first fluorescence filter, the passband range of the second fluorescence filter, and the passband range of the third fluorescence filter.

[0078] In some embodiments, in the second arrangement, the maximum passband wavelength of the first excitation light filter 212 and the second excitation light filter 222 is less than the minimum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322, the maximum passband wavelength of the third excitation light filter 232 is less than the minimum passband wavelength of the third fluorescence filter 332, and the minimum passband wavelength of the third excitation light filter 232 is greater than the maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322. Among them, the maximum passband wavelength of the first excitation light filter 212 is less than or equal to the maximum passband wavelength of the second excitation light filter 222. The maximum passband wavelength of the first fluorescence filter 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter 322.

[0079] Among them, in the second arrangement, it is preferred to arrange the third fluorescence filter 332 on a light path different from that of the first fluorescence filter 312 and the second fluorescence filter 322. The first fluorescence filter 312 and the second fluorescence filter 322 can be arranged on the same light path or different light paths. This is because the minimum passband wavelength of the third excitation light filter 232 is greater than the maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322, and the maximum passband wavelength of the third excitation light filter 232 is less than the minimum passband wavelength of the third fluorescence filter 332. When the third fluorescence filter 332 is arranged on the same light path as the first fluorescence filter 312 and / or the second fluorescence filter 322, when the first fluorescence, the second fluorescence and / or the third fluorescence enter the fluorescence detection channel, the third excitation light may also enter the fluorescence detection channel. At this time, setting a filter between the droplet detection flow channel 10 and the fluorescence detection module 30 cannot effectively filter the third excitation light, which will affect the detection result.

[0080] Further, on the optical path of the droplet detection channel 10 where the fluorescence detection module 30 is provided, between the droplet detection channel 10 and the fluorescence detection module 30, a filter element is also provided. For example, between the droplet detection channel 10 and the first fluorescence detection module 31, or between the droplet detection channel 10 and the second fluorescence detection module 32, a first filter element 61 is provided. The minimum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passbands of the first excitation light filter element 212 and the second excitation light filter element 222, and less than the minimum wavelength of the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322. The maximum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322, and less than the minimum wavelength of the passband of the third excitation light filter element 232. Between the droplet detection channel 10 and the third fluorescence detection module 33, a second filter element 62 is provided. The minimum wavelength of the passband of the second filter element 62 is less than the minimum wavelength of the passband of the third fluorescence filter element 332, and greater than the maximum wavelength of the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322. For example Figure 9 the first filter element 61 and the second filter element 62 in

[0081] In some embodiments, the passbands of the first excitation light filter element 212 and the second excitation light filter element 222 are separated from each other, as shown in Figure 12 ; or at least part of the passbands of the first excitation light filter element 212 and the second excitation light filter element 222 overlap, as shown in Figure 13 . In some embodiments, the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322 are separated from each other, as shown in Figure 12 ; or part of the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322 overlap, as shown in Figure 13 .

[0082] In some embodiments, the optical detection device 1 includes four excitation light modules 20 and four fluorescence detection modules 30. Specifically, as shown in Figures 14 - 15 :

[0083] The optical detection device 1 includes: a droplet detection flow channel 10, a first excitation light module 21, a second excitation light module 22, a third excitation light module 23, a fourth excitation light module 24, a first fluorescence detection module 31, a second fluorescence detection module 32, a third fluorescence detection module 33, and a fourth fluorescence detection module 34. Among them, the first excitation light module 21 includes a first excitation light emitter 211 and a first excitation light filter 212; the second excitation light module 22 includes a second excitation light emitter 221 and a second excitation light filter 222; the third excitation light module 23 includes a third excitation light emitter 231 and a third excitation light filter 232; the fourth excitation light module 24 includes a fourth excitation light emitter 241 and a fourth excitation light filter 242; the first fluorescence detection module 31 includes a first fluorescence detector 311 and a first fluorescence filter 312; the second fluorescence detection module 32 includes a second fluorescence detector 321 and a second fluorescence filter 322; the third fluorescence detection module 33 includes a third fluorescence detector 331 and a third fluorescence filter 332; the fourth fluorescence detection module 34 includes a fourth fluorescence detector 341 and a fourth fluorescence filter 342. The first excitation light emitter 211, the second excitation light emitter 221, the third excitation light emitter 231, and the fourth excitation light emitter 241 respectively emit a first excitation light, a second excitation light, a third excitation light, and a fourth excitation light. After the first excitation light, the second excitation light, the third excitation light, and the fourth excitation light are filtered by the first excitation light filter 212, the second excitation light filter 222, the third excitation light filter 232, and the fourth excitation light filter 242 respectively, they are converged and irradiated into the droplet detection flow channel 10, so that at least part of the droplets 11 to be detected flowing through the droplet detection flow channel 10 emit fluorescence accordingly. One path of fluorescence is received by the first fluorescence detector 311 after being filtered by the first fluorescence filter 312, one path of fluorescence is received by the second fluorescence detector 321 after being filtered by the second fluorescence filter 322, one path of fluorescence is received by the third fluorescence detector 331 after being filtered by the third fluorescence filter 332, and one path of fluorescence is received by the fourth fluorescence detector 341 after being filtered by the fourth fluorescence filter 342. At least part of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light passing through the first excitation light filter 212, the second excitation light filter 222, the third excitation light filter 232, and / or the fourth excitation light filter 242.

[0084] Exemplarily, see Figures 14 - 15 , Figure 14 which is a schematic structural diagram of a structure including four fluorescence detection modules 30 provided by an embodiment of the present application, Figure 15 which is a schematic structural diagram of a structure including four excitation light modules 20 provided by an embodiment of the present application. As Figure 14As shown, the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the first optical path of the droplet detection flow channel 10, and the third fluorescence detection module 33 and the fourth fluorescence detection module 34 are arranged on the second optical path of the droplet detection flow channel 10. The first optical path is different from the second optical path. As Figure 14 shown, the emitted light from the first excitation light module 21, the second excitation light module 22, the third excitation light module 23, and the fourth excitation light module 24 is converged onto the same optical path through a dichroic mirror, and is shaped by a spot shaping device 401 to form a spot and then irradiated on the droplet detection flow channel 10. It should be noted that, Figure 14 and Figure 15 This is only an example of the optical detection device 1 provided in this application. The first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, the fourth fluorescence detection module 34, the first excitation light module 21, the second excitation light module 22, the third excitation light module 23, and the fourth excitation light module 24 can of course also be assembled in other arrangements and with other components. This application does not make specific restrictions on this.

[0085] In the embodiments of this application, the arrangement order of the passband ranges of the filter elements includes the following three types:

[0086] The first type: the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, the fourth excitation light filter element 242, the first fluorescence filter element 312, the second fluorescence filter element 322, the third fluorescence filter element 332, the fourth fluorescence filter element 342.

[0087] The second type: the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, the first fluorescence filter element 312, the second fluorescence filter element 322, the third fluorescence filter element 332, the fourth excitation light filter element 242, the fourth fluorescence filter element 342.

[0088] The third type: the first excitation light filter element 212, the second excitation light filter element 222, the first fluorescence filter element 312, the second fluorescence filter element 322, the third excitation light filter element 232, the fourth excitation light filter element 242, the third fluorescence filter element 332, the fourth fluorescence filter element 342.

[0089] Specifically, in the first arrangement, the maximum passband wavelength of the first excitation light filter 212, the second excitation light filter 222, the third excitation light filter 232, and the fourth excitation light filter 242 is less than the minimum passband wavelength of the first fluorescence filter 312, the second fluorescence filter 322, the third fluorescence filter 332, and the fourth fluorescence filter 342. Among them, the maximum passband wavelength of the first excitation light filter 212 is less than or equal to the maximum passband wavelength of the second excitation light filter 222, which is less than or equal to the maximum passband wavelength of the third excitation light filter 232, which is less than or equal to the maximum passband wavelength of the fourth excitation light filter 242. The maximum passband wavelength of the first fluorescence filter 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter 322, which is less than or equal to the maximum passband wavelength of the third fluorescence filter 332, which is less than or equal to the maximum passband wavelength of the fourth fluorescence filter 342.

[0090] Among them, the first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, and the fourth fluorescence detection module 34 can be arranged on the same optical path of the droplet detection flow channel 10, or can be arranged on different optical paths of the droplet detection flow channel 10. When at least two of the first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, and the fourth fluorescence detection module 34 are arranged on the same optical path, a dichroic mirror is further included between two adjacent fluorescence detection modules 30. For example, a first dichroic mirror 51 is further arranged between the first fluorescence detection module 31 and the second fluorescence detection module 32, a second dichroic mirror is further arranged between the second fluorescence detection module 32 and the third fluorescence detection module 33, and a third dichroic mirror 52 is further included between the third fluorescence detection module 33 and the fourth fluorescence detection module 34. The cut-off wavelength of the first dichroic mirror 51 is greater than the minimum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322, and less than the maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322. The cut-off wavelength of the second dichroic mirror is greater than the minimum passband wavelength of the second fluorescence filter 322 and the third fluorescence filter 332, and less than the maximum passband wavelength of the second fluorescence filter 322 and the third fluorescence filter 332. The cut-off wavelength of the third dichroic mirror 52 is greater than the minimum passband wavelength of the third fluorescence filter 332 and the fourth fluorescence filter 342, and less than the maximum passband wavelength of the third fluorescence filter 332 and the fourth fluorescence filter 342. For example, see Figure 14Among them, the first fluorescence detection module 31 and the second fluorescence detection module 32 are arranged on the first optical path of the droplet detection flow channel 10, the third fluorescence detection module 33 and the fourth fluorescence detection module 34 are arranged on the second optical path of the droplet detection flow channel 10, a first dichroic mirror 51 is arranged between the first fluorescence detection module 31 and the second fluorescence detection module 32, the first dichroic mirror 51 is a long-pass dichroic mirror, the first fluorescence detection module 31 is arranged on the reflection optical path of the first dichroic mirror 51, and the second fluorescence detection module 32 is arranged on the transmission optical path of the first dichroic mirror 51. A third dichroic mirror 52 is arranged between the third fluorescence detection module 33 and the fourth fluorescence detection module 34, the third dichroic mirror 52 is a long-pass dichroic mirror, the third fluorescence detection module 33 is arranged on the reflection optical path of the third dichroic mirror 52, and the fourth fluorescence detection module 34 is arranged on the transmission optical path of the second dichroic mirror. Of course, the first dichroic mirror 51 and the third dichroic mirror 52 can also be short-pass dichroic mirrors, and the specific setting method will not be described in detail here.

[0091] Further, on the optical path of the droplet detection flow channel 10 where the fluorescence detection module 30 is arranged, between the droplet detection flow channel 10 and the fluorescence detection module 30, a first filter element 61 is further arranged. The minimum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passbands of the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, and the fourth excitation light filter element 242, and less than the minimum wavelength of the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, the third fluorescence filter element 332, and the fourth fluorescence filter element 342. For example, see Figure 14 the first filter element 61 in

[0092] In some embodiments, the passbands of the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, and the fourth excitation light filter element 242 are separated from each other; or there is partial or complete overlap among the passbands of the first excitation light filter element 212, the second excitation light filter element 222, the third excitation light filter element 232, and the third excitation light filter element 232.

[0093] In some embodiments, the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, the third fluorescence filter element 332, and the fourth fluorescence filter element 342 are separated from each other; or there is partial or complete overlap among the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, the third fluorescence filter element 332, and the third fluorescence filter element 332.

[0094] Specifically, in the second arrangement, the maximum passband wavelength of the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232 is less than the minimum passband wavelength of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332, and the maximum passband wavelength of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332 is less than the minimum passband wavelength of the fourth excitation light filter 242. The maximum passband wavelength of the first excitation light filter 212 is less than or equal to the maximum passband wavelength of the second excitation light filter 222 which is less than or equal to the maximum passband wavelength of the third excitation light filter 232. The maximum passband wavelength of the first fluorescence filter 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter 322 which is less than or equal to the maximum passband wavelength of the third fluorescence filter 332. The maximum passband wavelength of the fourth excitation light filter 242 is less than the minimum passband wavelength of the fourth fluorescence filter 342.

[0095] Among them, the first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, and the fourth fluorescence detection module 34 can be arranged on the same optical path of the droplet detection flow channel 10, or can be arranged on different optical paths of the droplet detection flow channel 10.

[0096] Among them, it is preferably to arrange the fourth fluorescence filter 342 on an optical path different from that of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332. The first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332 can be arranged on the same optical path or different optical paths. This is because the minimum passband wavelength of the fourth excitation light filter 242 is greater than the maximum passband wavelength of the first fluorescence filter 312, the second fluorescence filter 322, and the third fluorescence filter 332, and the maximum passband wavelength of the fourth excitation light filter 242 is less than the minimum passband wavelength of the fourth fluorescence filter 342. When the fourth fluorescence filter 342 is arranged on the same optical path as the first fluorescence filter 312, the second fluorescence filter 322, and / or the third fluorescence filter 332, when the first fluorescence, the second fluorescence, the third fluorescence, and / or the fourth fluorescence enter the fluorescence detection channel, the fourth excitation light may also enter the fluorescence detection channel. At this time, even if a filter is arranged between the droplet detection flow channel 10 and the fluorescence detection module 30, the fourth excitation light cannot be effectively filtered, affecting the detection result.

[0097] Further, on the optical path of the fluorescence detection module 30 provided in the droplet detection channel 10, between the droplet detection channel 10 and the fluorescence detection module 30, a filter element is also provided. For example, between the droplet detection channel 10 and the first fluorescence detection module 31, or between the droplet detection channel 10 and the second fluorescence detection module 32, or between the droplet detection channel 10 and the third fluorescence detection module 33, a first filter element 61 is provided. The minimum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passbands of the first excitation light filter element 212, the second excitation light filter element 222, and the third excitation light filter element 232, and less than the minimum wavelength of the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332. The maximum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332, and less than the minimum wavelength of the passband of the fourth excitation light filter element 242. Between the droplet detection channel 10 and the fourth fluorescence detection module 34, a second filter element 62 is provided. The minimum wavelength of the passband of the second filter element 62 is less than the minimum wavelength of the passband of the fourth fluorescence filter element 342, and greater than the maximum wavelength of the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332. For example, see Figure 14 the first filter element 61 and the second filter element 62 in

[0098] In some embodiments, the passbands of the first excitation light filter element 212, the second excitation light filter element 222, and the third excitation light filter element 232 are separated from each other; or there is partial or complete overlap among the passbands of the first excitation light filter element 212, the second excitation light filter element 222, and the third excitation light filter element 232.

[0099] In some embodiments, the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332 are separated from each other; or there is partial or complete overlap among the passbands of the first fluorescence filter element 312, the second fluorescence filter element 322, and the third fluorescence filter element 332.

[0100] Specifically, in the third arrangement, the maximum passband wavelength of the first excitation light filter 212 and the second excitation light filter 222 is less than the minimum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322. The maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322 is less than the minimum passband wavelength of the third excitation light filter 232 and the fourth excitation light filter 242. The maximum passband wavelength of the third excitation light filter 232 and the fourth excitation light filter 242 is less than the minimum passband wavelength of the third fluorescence filter 332 and the fourth fluorescence filter 342. Among them, the maximum passband wavelength of the first excitation light filter 212 is less than or equal to the maximum passband wavelength of the second excitation light filter 222, the maximum passband wavelength of the first fluorescence filter 312 is less than or equal to the maximum passband wavelength of the second fluorescence filter 322, the maximum passband wavelength of the third excitation light filter 232 is less than or equal to the maximum passband wavelength of the fourth excitation light filter 242, and the maximum passband wavelength of the third fluorescence filter 332 is less than or equal to the maximum passband wavelength of the fourth fluorescence filter 342.

[0101] Among them, the first fluorescence detection module 31, the second fluorescence detection module 32, the third fluorescence detection module 33, and the fourth fluorescence detection module 34 can be arranged on the same optical path of the droplet detection channel 10 or on different optical paths of the droplet detection channel 10.

[0102] Among them, in the third arrangement, it is preferable to arrange the third fluorescence filter 332 and the fourth fluorescence filter 342 on optical paths different from those of the first fluorescence filter 312 and the second fluorescence filter 322. The first fluorescence filter 312 and the second fluorescence filter 322 can be arranged on the same optical path or different optical paths. The third fluorescence filter 332 and the fourth fluorescence filter 342 can be arranged on the same optical path or different optical paths. This is because the minimum passband wavelength of the third excitation light filter 232 and the fourth excitation light filter 242 is greater than the maximum passband wavelength of the first fluorescence filter 312 and the second fluorescence filter 322, and the maximum passband wavelength of the third excitation light filter 232 and the fourth excitation light filter 242 is less than the minimum passband wavelength of the third fluorescence filter 332 and the fourth fluorescence filter 342. When the third fluorescence filter 332 and the fourth fluorescence filter 342 are arranged on the same optical path as the first fluorescence filter 312 and / or the second fluorescence filter 322, when the first fluorescence, the second fluorescence, the third fluorescence, and / or the fourth fluorescence enter the fluorescence detection channel, the third excitation light and / or the fourth excitation light may also enter the fluorescence detection channel. At this time, even if a filter is arranged between the droplet detection channel 10 and the fluorescence detection module 30, it is impossible to effectively filter the third excitation light and / or the fourth excitation light, affecting the detection result.

[0103] Further, on the optical path of the fluorescence detection module 30 provided in the droplet detection channel 10, between the droplet detection channel 10 and the fluorescence detection module 30, a filter element is also provided. For example, between the droplet detection channel 10 and the first fluorescence detection module 31, or between the droplet detection channel 10 and the second fluorescence detection module 32, a first filter element 61 is provided. The minimum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passband of the first excitation light filter element 212 and the second excitation light filter element 222, and less than the minimum wavelength of the passband of the first fluorescence filter element 312 and the second fluorescence filter element 322. The maximum wavelength of the passband of the first filter element 61 is greater than the maximum wavelength of the passband of the first fluorescence filter element 312 and the second fluorescence filter element 322, and less than the minimum wavelength of the passband of the third excitation light filter element 232 and the fourth excitation light filter element 242. Between the droplet detection channel 10 and the fourth fluorescence detection module 34, or between the droplet detection channel 10 and the third fluorescence detection module 33, a second filter element 62 is provided. The minimum wavelength of the passband of the second filter element 62 is less than the minimum wavelength of the passband of the third fluorescence filter element 332 and the fourth fluorescence filter element 342, and greater than the maximum wavelength of the passband of the first fluorescence filter element 312 and the second fluorescence filter element 322. For example, see Figure 14 the first filter element 61 and the second filter element 62 in

[0104] In some embodiments, the passbands of the first excitation light filter element 212 and the second excitation light filter element 222 are separated from each other; or the passband of the first excitation light filter element and the passband of the second excitation light filter element 222 partially or completely overlap. In some embodiments, the passbands of the third excitation light filter element 232 and the fourth excitation light filter element 242 are separated from each other; or the passband of the third excitation light filter element 232 and the passband of the fourth excitation light filter element 242 partially or completely overlap.

[0105] In some embodiments, the passbands of the first fluorescence filter element 312 and the second fluorescence filter element 322 are separated from each other; or the passband of the first excitation light filter element and the passband of the second fluorescence filter element 322 partially or completely overlap. In some embodiments, the passbands of the third fluorescence filter element 332 and the fourth fluorescence filter element 342 are separated from each other; or the passband of the third fluorescence filter element 332 and the passband of the fourth fluorescence filter element 342 partially or completely overlap.

[0106] It should be noted that in addition to the cases including 2 - 4 fluorescence detection modules 30 and excitation light modules 20 listed in the above embodiments, the optical detection device 1 provided in the embodiments of the present application may further include a greater number of fluorescence detection modules 30 and excitation light modules 20. The specific setting method of the arrangement order of the passband ranges in multiple excitation light filter elements and multiple fluorescence filter elements refers to the methods of the above embodiments, and the present application will not list them one by one.

[0107] In this application, the mention of "embodiment" or "implementation manner" means that the specific features, structures or characteristics described in connection with the embodiments can be included in at least one embodiment of this application. The appearance of the phrase at various positions in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. Those skilled in the art will explicitly and implicitly understand that the embodiments described in this application can be combined with other embodiments. In addition, it should also be understood that the features, structures or characteristics described in the various embodiments of this application can be combined arbitrarily without contradiction to form another embodiment that does not depart from the spirit and scope of the technical solution of this application.

[0108] Finally, it should be noted that the above implementation manners are only used to illustrate the technical solutions of this application and not to limit them. Although the technical solutions of this application have been described in detail with reference to the above preferred implementation manners, those of ordinary skill in the art should understand that modifications or equivalent replacements can be made to the technical solutions of this application without departing from the spirit and scope of the technical solutions of this application.

Claims

1. An optical detection device for polymerase chain reaction, characterized in that: include: A droplet detection channel, for allowing a fluid to be detected to flow through, wherein the fluid to be detected at least includes droplets to be detected; A first excitation light module, comprising a first excitation light emitter and a first excitation light filter; A second excitation light module, comprising a second excitation light emitter and a second excitation light filter; A first fluorescence detection module, comprising a first fluorescence detector and a first fluorescence filter; A second fluorescence detection module, comprising a second fluorescence detector and a second fluorescence filter; The first excitation light emitter and the second excitation light emitter emit a first excitation light and a second excitation light respectively, and the first excitation light and the second excitation light are respectively filtered by the first excitation light filter and the second excitation light filter, and then converged and irradiated into the droplet detection channel, so that at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, wherein one path of the fluorescence is filtered by the first fluorescence filter and then received by the first fluorescence detector, and the other path of the fluorescence is filtered by the second fluorescence filter and then received by the second fluorescence detector; The maximum passband wavelength of the first excitation light filter and the second excitation light filter is smaller than the minimum passband wavelength of the first fluorescence filter and the second fluorescence filter.

2. The optical detection device according to claim 1, characterized in that: The passband of the first excitation light filter and the passband of the second excitation light filter are separated from each other; or the passband of the first excitation light filter and the passband of the second excitation light filter partially overlap.

3. The optical detection device according to claim 1, characterized in that: The passband of the first fluorescent filter and the passband of the second fluorescent filter are separated from each other; or the passband of the first fluorescent filter and the passband of the second fluorescent filter partially overlap.

4. The optical detection device according to claim 1, characterized in that: It also includes a third excitation light module and a third fluorescence detection module; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; The third excitation light emitter emits a third excitation light, and after passing through the third excitation light filter, the third excitation light converges with the first excitation light and the second excitation light and irradiates the droplet detection channel, so that at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, wherein one path of the fluorescence is filtered by the third fluorescence filter and received by the third fluorescence detector; The maximum passband wavelength of the first excitation light filter, the second excitation light filter and the third excitation light filter is smaller than the minimum passband wavelength of the first fluorescence filter, the second fluorescence filter and the third fluorescence filter.

5. The optical detection device according to claim 1, characterized in that: It also includes a third excitation light module and a third fluorescence detection module; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; The third excitation light emitter emits a third excitation light, and after passing through the third excitation light filter, the third excitation light converges with the first excitation light and the second excitation light and irradiates the droplet detection channel, so that at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, wherein one path of the fluorescence is filtered by the third fluorescence filter and received by the third fluorescence detector; Among them, the maximum passband wavelength of the third excitation light filter is smaller than the minimum passband wavelength of the third fluorescence filter, and the minimum passband wavelength of the third excitation light filter is larger than the maximum passband wavelengths of the first fluorescence filter and the second fluorescence filter.

6. The optical detection device according to claim 1, characterized in that: It also includes a third excitation light module, a third fluorescence detection module, a fourth excitation light module and a fourth fluorescence detection module; the third excitation light module includes a third excitation light emitter and a third excitation light filter; the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter; the fourth excitation light module includes a fourth excitation light emitter and a fourth excitation light filter; the fourth fluorescence detection module includes a fourth fluorescence detector and a fourth fluorescence filter; The third excitation light emitter and the fourth excitation light emitter emit a third excitation light and a fourth excitation light respectively. After the third excitation light and the fourth excitation light pass through the third excitation light filter and the fourth excitation light filter respectively, the first excitation light, the second excitation light, the third excitation light and the fourth excitation light converge and irradiate the droplet detection channel, so that at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, wherein one path of the fluorescence is received by the third fluorescence detector after being filtered by the third fluorescence filter, and one path of the fluorescence is received by the fourth fluorescence detector after being filtered by the fourth fluorescence filter; Among them, the minimum passband wavelength in the third excitation light filter and the fourth excitation light filter is greater than the maximum passband wavelength in the first fluorescence filter and the second fluorescence filter, and the maximum passband wavelength in the third excitation light filter and the fourth excitation light filter is smaller than the minimum passband wavelength in the third fluorescence filter and the fourth fluorescence filter.

7. The optical detection device according to claim 6, characterized in that: The passband of the third excitation light filter and the passband of the fourth excitation light filter are separated from each other; or the passband of the third excitation light filter and the passband of the fourth excitation light filter partially overlap.

8. The optical detection device according to claim 6, characterized in that: The passband of the third fluorescent filter and the passband of the fourth fluorescent filter are separated from each other; or the passband of the third fluorescent filter and the passband of the fourth fluorescent filter partially overlap.

9. The optical detection device according to claim 1, characterized in that: It also includes a first filter; the droplet to be detected emits fluorescence, and after being filtered by the first filter, one path of the fluorescence is filtered by the first fluorescence filter and received by the first fluorescence detector, and the other path of the fluorescence is filtered by the second fluorescence filter and received by the second fluorescence detector, wherein the maximum wavelength of the passband in the first excitation light filter and the second excitation light filter is less than the minimum wavelength of the passband of the first filter; and / or It also includes a first dichroic mirror, the first fluorescence detector and the first fluorescence filter are both arranged on the transmission optical path of the first dichroic mirror, the second fluorescence detector and the second fluorescence filter are both arranged on the reflection optical path of the first dichroic mirror, wherein the cutoff wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband of the first fluorescence filter and the second fluorescence filter, and is smaller than the maximum wavelength of the passband of the first fluorescence filter and the second fluorescence filter.

10. The optical detection device according to claim 6, characterized in that: It also includes a second filter; the droplet to be detected emits fluorescence in response, and after being filtered by the second filter, one path of the fluorescence is filtered by the third fluorescence filter and then received by the third fluorescence detector, and the other path of the fluorescence is filtered by the fourth fluorescence filter and then received by the fourth fluorescence detector, wherein the maximum wavelength of the passband in the third excitation light filter and the fourth excitation light filter is less than the minimum wavelength of the passband of the second filter; and / or Also comprising a second dichroic mirror, the third fluorescence detector and the third fluorescence filter are both arranged on the transmission optical path of the second dichroic mirror, and the fourth fluorescence detector and the fourth fluorescence filter are both arranged on the reflection optical path of the second dichroic mirror; The cutoff wavelength of the second dichroic mirror is greater than the minimum wavelength of the passband of the third fluorescent filter and the fourth fluorescent filter, and is smaller than the maximum wavelength of the passband of the third fluorescent filter and the fourth fluorescent filter.

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