Microdroplet fluorescence detection device

Abstract

The embodiment of the invention provides a micro-droplet fluorescence detection device, which comprises a micro-droplet detection flow channel for a to-be-detected fluid to flow through, and the to-be-detected fluid at least comprises to-be-detected micro-droplets and bubbles; the bubble detection module comprises a bubble detector and a first dichroscope, and the bubble detector and the first fluorescence detector are respectively arranged on a transmission optical path or a reflection optical path of the first dichroscope; the first exciting light emitter emits exciting light to irradiate the droplet detection flow channel, at least part of to-be-detected droplets flowing through the droplet detection flow channel correspondingly emit fluorescent light, or bubbles flowing through the droplet detection flow channel correspondingly reflect and / or scatter part of exciting light; the first dichroscope sends the transmitted or reflected exciting light to the bubble detector, so that a bubble signal is detected, and the bubble signal is removed from a fluorescence detection result, so that the accuracy of the detection result can be improved.

Description

Technical Field

[0001] This application relates to the field of optical detection technologies, and in particular, to a droplet fluorescence detection device. Background Art

[0002] Droplet digital PCR (ddPCR) belongs to a type of digital PCR. Through an emulsification technique, a sample is divided into tens of thousands of "water-in-oil" droplets, and the signals of the droplets are analyzed by a flow cytometry fluorescence method. In a "water-in-oil" emulsion, abnormal signals caused by bubbles and the like often affect the accuracy of test results. How to accurately identify and eliminate abnormal signals in the emulsion has become a major technical problem in ddPCR. Summary of the Invention

[0003] An embodiment of this application provides a droplet fluorescence detection device. A bubble detection module is arranged in the droplet fluorescence detection device to effectively detect bubble signals and eliminate the bubble signals, thereby improving the accuracy of detection results.

[0004] An embodiment of this application provides a droplet fluorescence detection device. The droplet fluorescence detection device includes: a droplet detection flow channel for allowing a fluid to be detected to flow through, where the fluid to be detected at least includes droplets to be detected and bubbles; a first excitation light module including a first excitation light emitter; a first fluorescence detection module including a first fluorescence detector; a bubble detection module including a bubble detector; a first dichroic mirror, where the bubble detector and the first fluorescence detector are respectively arranged on the transmission optical path or the reflection optical path of the first dichroic mirror; the first excitation light emitter emits excitation light that irradiates the droplet detection flow channel. Under the irradiation of the excitation light, at least some of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence accordingly, or the bubbles flowing through the droplet detection flow channel reflect and / or scatter part of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror. The first dichroic mirror transmits one of the fluorescence or the excitation light and reflects the other. The fluorescence that has passed through transmission or reflection is received by the first fluorescence detector, and the excitation light that has passed through transmission or reflection is received by the bubble detector.

[0005] In some embodiments, the first excitation light module further includes a first excitation light filter. The first excitation light emitter emits excitation light, which, after being filtered by the first excitation light filter, irradiates on the droplet detection flow channel. The first fluorescence detection module further includes a first fluorescence filter. The fluorescence that is transmitted or reflected by the first dichroic mirror is received by the first fluorescence detector after being filtered by the first fluorescence filter. Both the first fluorescence detector and the first fluorescence filter are disposed on the transmission optical path of the first dichroic mirror, and the bubble detector is disposed on the reflection optical path of the first dichroic mirror. The cut-off wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband of the first excitation light filter and less than the minimum wavelength of the passband of the first fluorescence filter.

[0006] In some embodiments, the first excitation light module further includes a first excitation light filter. The first excitation light emitter emits excitation light, which, after being filtered by the first excitation light filter, irradiates on the droplet detection flow channel. The first fluorescence detection module further includes a first fluorescence filter. The fluorescence that is transmitted or reflected by the first dichroic mirror is received by the first fluorescence detector after being filtered by the first fluorescence filter. Both the first fluorescence detector and the first fluorescence filter are disposed on the reflection optical path of the first dichroic mirror, and the bubble detector is disposed on the transmission optical path of the first dichroic mirror. The cut-off wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband of the first excitation light filter and less than the minimum wavelength of the passband of the first fluorescence filter.

[0007] In some embodiments, the bubble detector includes a photodiode PD, an avalanche photodiode APD, a multi-pixel photon counter MPPC, or a photomultiplier tube PMT.

[0008] In some embodiments, the first fluorescence detector receives the fluorescence synchronously with the bubble detector receiving the excitation light.

[0009] In some embodiments, it further includes at least one of a second excitation light module, a third excitation light module, and a fourth excitation light module, and correspondingly includes at least one of a second fluorescence detection module, a third fluorescence detection module, and a fourth fluorescence detection module; the second excitation light module includes a second excitation light emitter and a second excitation light filter, and the second fluorescence detection module includes a second fluorescence detector and a second fluorescence filter. Among them, the second excitation light emitter emits excitation light, which, after passing through the second excitation light filter, irradiates on the droplet detection flow channel. Under the irradiation of the excitation light, at least some of the droplets to be detected flowing through the droplet detection flow channel correspondingly emit fluorescence. After the fluorescence is filtered by the second fluorescence filter, it is received by the second fluorescence detector; the third excitation light module includes a third excitation light emitter and a third excitation light filter, and the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter. Among them, the third excitation light emitter emits excitation light, which, after passing through the third excitation light filter, irradiates on the droplet detection flow channel. Under the irradiation of the excitation light, at least some of the droplets to be detected flowing through the droplet detection flow channel correspondingly emit fluorescence. After the fluorescence is filtered by the third fluorescence filter, it is received by the third fluorescence detector; the fourth excitation light module further includes a fourth excitation light emitter and a fourth excitation light filter, and the fourth fluorescence detection module includes a fourth fluorescence detector and a fourth fluorescence filter. Among them, the fourth excitation light emitter emits excitation light, which, after passing through the fourth excitation light filter, irradiates on the droplet detection flow channel. Under the irradiation of the excitation light, at least some of the droplets to be detected flowing through the droplet detection flow channel correspondingly emit fluorescence. After the fluorescence is filtered by the fourth fluorescence filter, it is received by the fourth fluorescence detector.

[0010] In some embodiments, it includes a second excitation light module and the second fluorescence detection module; it further includes a second dichroic mirror, and the second dichroic mirror is arranged on the transmission optical path or the reflection optical path of the first dichroic mirror; the first fluorescence detector and the first fluorescence filter are arranged on one of the transmission optical path or the reflection optical path of the second dichroic mirror, and the second fluorescence detector and the second fluorescence filter are arranged on the other of the transmission optical path or the reflection optical path of the second dichroic mirror.

[0011] In some embodiments, the maximum wavelength of the passband of the first excitation light filter is less than the maximum wavelength of the passband of the second excitation light filter, and the maximum wavelength of the passband of the first fluorescence filter is less than the maximum wavelength of the passband of the second fluorescence filter.

[0012] In some embodiments, it includes a third excitation light module, a fourth excitation light module, a third fluorescence detection module, and the fourth fluorescence detection module; it further includes a third dichroic mirror, and the third dichroic mirror is disposed on the transmission optical path or the reflection optical path of the first dichroic mirror; the third fluorescence detector and the third fluorescence filter are disposed on one of the transmission optical path or the reflection optical path of the third dichroic mirror, and the fourth fluorescence detector and the fourth fluorescence filter are disposed on the other of the transmission optical path or the reflection optical path of the third dichroic mirror; wherein, the maximum wavelength of the passband of the third excitation light filter is less than the maximum wavelength of the passband of the fourth excitation light filter, and the maximum wavelength of the passband of the third fluorescence filter is less than the maximum wavelength of the passband of the fourth fluorescence filter.

[0013] In some embodiments, it further includes a processor, and the processor is configured to receive the fluorescence signal collected by the first fluorescence detector and the bubble signal collected by the bubble detector, determine the fluorescence signal synchronized with the bubble signal as an invalid fluorescence signal, and after removing the invalid fluorescence signal from the fluorescence signal, determine the detection result of the fluid to be detected according to the fluorescence signal after removal.

[0014] In the embodiments of the present application, the droplet fluorescence detection device includes: a droplet detection flow channel for allowing the fluid to be detected to flow through, wherein the fluid to be detected at least includes droplets to be detected and bubbles; a first excitation light module including a first excitation light emitter; a first fluorescence detection module including a first fluorescence detector; a bubble detection module including a bubble detector and a first dichroic mirror, and the bubble detector and the first fluorescence detector are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror; the first excitation light emitter emits excitation light to irradiate on the droplet detection flow channel, and under the irradiation of the excitation light, at least part of the droplets to be detected flowing through the droplet detection flow channel emit fluorescence correspondingly, or the bubbles flowing through the droplet detection flow channel reflect and / or scatter part of the excitation light correspondingly, the emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror, the first dichroic mirror transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light, the fluorescence passing through transmission or reflection is received by the first fluorescence detector, and the excitation light passing through transmission or reflection is received by the bubble detector, thereby detecting a bubble signal, and by removing the bubble signal from the fluorescence detection result, the accuracy of the detection result can be improved. Description of the Drawings

[0015] To more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the accompanying drawings required for the description of the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can be obtained based on these drawings.

[0016] Figure 1 Schematic diagram of the excitation light spectrum and emission light spectrum of the fluorescent dye provided by the embodiment of the present application;

[0017] Figure 2 Block diagram of the structure of a droplet fluorescence detection device provided by the embodiment of the present application;

[0018] Figure 3 Schematic diagram of the structure of an excitation light module provided by the embodiment of the present application;

[0019] Figure 4 Schematic diagram of the structure of a fluorescence detection module provided by the embodiment of the present application;

[0020] Figure 5 Schematic diagram of the structure of two excitation light modules provided by the embodiment of the present application;

[0021] Figure 6 Schematic diagram of the structure of two fluorescence detection modules provided by the embodiment of the present application;

[0022] Figure 7 Schematic diagram of the structure of three excitation light modules provided by the embodiment of the present application;

[0023] Figure 8 Schematic diagram of the structure of three fluorescence detection modules provided by the embodiment of the present application;

[0024] Figure 9 Schematic diagram of the structure of four excitation light modules provided by the embodiment of the present application;

[0025] Figure 10 Schematic diagram of the structure of four fluorescence detection modules provided by the embodiment of the present application;

[0026] Figure 11 Schematic diagram of the changes in fluorescence signals and bubble signals when only the droplet to be detected passes through, provided by the embodiment of the present application;

[0027] Figure 12 Schematic diagram of the changes in fluorescence signals and bubble signals when multiple consecutive bubbles pass through, provided by the embodiment of the present application. Detailed implementation manners

[0028] To enable those skilled in the art to better understand the solution of this application, the technical solution in this application will be clearly and completely described below in conjunction with the accompanying drawings in this application. Obviously, the described embodiments are only a part of the embodiments of this application, rather than all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of this application.

[0029] 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 with a longer wavelength is generated, namely fluorescence. As Figure 1 shown is the excitation light spectrum and emission light spectrum of a fluorescent dye. Different fluorescent dyes have different excitation light wavelengths and emission light wavelengths, and have different excitation efficiencies and emission efficiencies at different wavelengths.

[0030] In the actual fluorescence detection process, the droplet digital PCR (ddPCR) technology and the emulsification technology can be used to form numerous individual emulsified droplets from the sample to be detected. The emulsified droplets are usually "oil-in-water" or "water-in-oil" droplets. Some of the emulsified droplets contain target nucleic acid molecules (denoted as positive droplets), and some do not (denoted as negative droplets). The types and / or concentrations of the fluorescent dyes contained in the positive droplets and negative droplets are different. By setting the excitation light to excite each emulsified droplet to generate fluorescence, and then detecting the type and / or intensity of the fluorescence through a fluorescence detector, the number of positive droplets and negative droplets can be detected, and then the detection result of the fluid to be detected can be determined. However, abnormal signals such as air bubbles in the emulsion often affect the accuracy of the test results. How to accurately identify and eliminate the abnormal signals in the emulsion has become a major technical problem in ddPCR.

[0031] To solve the above technical problems, this application provides a droplet fluorescence detection device, which is provided with a bubble detection module to effectively detect the bubble signal and eliminate the bubble signal, thereby improving the accuracy of the detection result. For the droplet fluorescence detection device provided by the embodiments of this application, please refer to the following content and Figures 2 - 12 :

[0032] See Figure 2, the droplet fluorescence detection device 1 includes a droplet detection flow channel 10, at least one excitation light module 20, at least one fluorescence detection module 30, and a bubble detection module 40. The droplet detection flow channel 10 is used for the fluid to be detected to flow through, where the fluid to be detected at least includes droplets to be detected 11 and bubbles, and the droplets to be detected 11 include at least one fluorescent dye. The excitation light module 20 is used to generate excitation light and excite the fluorescent dye in the droplets to be detected 11 to generate fluorescence. The fluorescence detection module 30 is used to receive the fluorescence to obtain the detection data of the droplets to be detected 11. The bubble detection module 40 is used to detect the bubble situation in the fluid to be detected.

[0033] 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 droplet fluorescence detection device 1 may include one excitation light module 20 and one fluorescence detection module 30; for example, the droplet fluorescence detection device 1 may include four excitation light modules 20 and four fluorescence detection modules 30; of course, it may also include any number of the same excitation light modules 20 and fluorescence detection modules 30.

[0034] 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 modulation. 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. Preferably, 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.

[0035] 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.

[0036] In some embodiments, the droplet fluorescence detection device 1 further includes a spot shaping device 601, see Figure 3 , 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.

[0037] In some embodiments, when the droplet fluorescence detection device 1 includes more than two excitation light modules 20, the droplet fluorescence detection device 1 further includes a dichroic mirror 602, see Figure 5 , which is used to converge the excitation light generated by multiple excitation light modules 20 on an optical path, thereby saving the space resources of the droplet fluorescence detection device 1.

[0038] In some embodiments, at least one excitation light module 20 further includes a brightness feedback module 603. See Figure 3 , the brightness feedback module 603 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 603 and detected by the brightness detector. The brightness detector feeds back the detected brightness of the excitation light to the processor 50, and the processor 50 then controls the brightness of the excitation light generated by the excitation light emitter in a closed loop. In some embodiments, the brightness feedback module 603 further includes a filter element and a converging lens.

[0039] In some embodiments, the fluorescence detection module 30 includes a fluorescence detector and a fluorescence filter element. The droplet 11 to be detected 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 element and then received by the fluorescence detector.

[0040] Among them, the fluorescence detection module 30 can be flexibly arranged on any side of the droplet detection channel 10. In some embodiments, when the droplet fluorescence detection device 1 includes more than two fluorescence detection modules 30, the more than two fluorescence detection modules 30 can be arranged on the same side of the droplet detection channel 10 at the same time, or the more than two fluorescence detection modules 30 can be respectively arranged on each side of the droplet detection channel 10.

[0041] In some embodiments, between the fluorescence detection module 30 and the droplet detection channel 10, there is further at least one first filter element 91 for filtering the excitation light to prevent the excitation light from entering the fluorescence detection module 30 and thus affecting the detection result. In some embodiments, when a fluorescence detection module 30 is arranged on the same side of the droplet detection channel 10, one first filter element 91 is included. Exemplarily, see Figure 4 , a first filter element 91 is arranged between the first dichroic mirror 71 and the first fluorescence detection module 31; in some embodiments, when at least two fluorescence detection modules 30 are respectively arranged on both sides of the droplet detection channel 10, two first filter elements 91 are included. Exemplarily, see Figure 10 , a first filter element 91 is further arranged between the second dichroic mirror 72 and the first dichroic mirror 71, and a first filter element 91 is further included between the droplet detection channel 10 and the third dichroic mirror.

[0042] In some embodiments, the bubble detection module 40 includes a bubble detector 41. See Figure 4 , the bubbles flowing through the droplet detection channel 10 will reflect and / or scatter part of the excitation light under the irradiation of the excitation light, and this part of the excitation light reflected and / or scattered is received by the bubble detector 41, so as to detect the bubble condition in the fluid to be detected.

[0043] Among them, the bubble detection module 40 can be flexibly arranged on both sides of the microdroplet detection flow channel 10. In some embodiments, the bubble detection module 40 and the fluorescence detection module 30 are arranged on the same side of the microdroplet detection flow channel 10, as shown in Figure 4 . In some other embodiments, at least one fluorescence detection module 30 is arranged on the first side of the microdroplet detection flow channel 10, and the bubble detection module 40 is arranged on the second side of the microdroplet detection flow channel 10, where the first side is different from the second side.

[0044] In some embodiments, the bubble detection module 40 and the fluorescence detection module 30 are arranged on the same side of the microdroplet detection flow channel 10, and the microdroplet fluorescence detection device 1 further includes a first dichroic mirror 71, as shown in Figure 4 . 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 with a wavelength greater than its cut-off wavelength, and to reflect light with a wavelength less than its cut-off wavelength. The short-pass dichroic mirror is used to pass light with a wavelength less than its cut-off wavelength, and to reflect light with a wavelength less than its cut-off wavelength. Herein, the optical path of the transmitted light is hereinafter referred to as the transmission optical path, and the optical path of the reflected light is referred to as the reflection optical path. In the embodiments of the present application, the first dichroic mirror 71 can be either a long-pass dichroic mirror or a short-pass dichroic mirror.

[0045] In some embodiments, the bubble detector 41 includes a photodiode PD, an avalanche photodiode APD, a multi-pixel photon counter MPPC, or a photomultiplier tube PMT.

[0046] Among them, the photodiode (Photo Diode, PD) belongs to a semiconductor photodetector. When a part of the excitation light reflected and / or scattered by the bubble irradiates on the photodiode, the photon energy is absorbed, causing the electrons in the photodiode to transition and generating a photocurrent. In some embodiments, the presence of bubbles is judged by detecting the change of the photocurrent. When a bubble passes by, the intensity of the reflected and / or scattered excitation light changes, and the photocurrent also changes accordingly. By monitoring and analyzing the photocurrent, the presence of bubbles and their related information, such as the size or number of bubbles, can be detected.

[0047] Among them, an Avalanche Photo Diode (APD) belongs to semiconductor photodetectors. The generation mechanism of the photocurrent of the APD is the same as that of the PD. By applying a reverse voltage to the PN junction, under the reverse bias voltage, when the excitation light reflected and / or scattered by the bubble irradiates the avalanche photodiode, the initial carriers generated by the photons obtain sufficient energy under the action of a high electric field, collide with lattice atoms to generate new carriers, and these new carriers continue to collide to generate more carriers, forming an avalanche multiplication effect, which greatly amplifies the photocurrent. In some embodiments, by detecting the change in the photocurrent after avalanche multiplication, the excitation light reflected and / or scattered by the bubble can be detected, thereby discovering the existence of the bubble; the advantage of the avalanche photodiode is that it has a high gain and a fast response speed, and can quickly and sensitively detect the change in the bubble reflection light signal.

[0048] Among them, a Multi-Pixel Photon Counter (MPPC) belongs to semiconductor photodetectors. The MPPC is a photon counting device composed of multiple APDs operating in Geiger mode. In Geiger mode, the reverse voltage of the APD is higher than the breakdown voltage. When the excitation light reflected and / or scattered by the bubble irradiates the MPPC, the semiconductor material inside it absorbs the photon energy to generate electron-hole pairs, and the APDs in the MPPC are in a reverse bias state close to avalanche breakdown, so that the generated electron-hole pairs are accelerated under the action of a strong electric field and trigger impact ionization, generating an avalanche multiplication effect to amplify the electrical signal. Finally, the circuit in the MPPC processes the electrical pulse signals generated by each pixel unit, and obtains information such as the intensity and number of photons of the excitation light by statistically analyzing the counts of each pixel unit. In some embodiments, the relevant situation of the bubble can be judged according to information such as the intensity or the number of photons of the excitation light.

[0049] Among them, a Photomultiplier Tube (PMT) belongs to vacuum electron devices. The excitation light reflected and / or scattered by the bubble is incident on the photocathode of the photomultiplier tube, and the photocathode emits photoelectrons after absorbing photons. These photoelectrons are accelerated under the action of an electric field and hit the dynodes. Each photoelectron generates multiple secondary electrons when hitting the dynode. After the cascade amplification of multiple dynodes, a detectable electrical signal is finally formed on the anode. In some embodiments, based on the very high sensitivity and gain of the photomultiplier tube, it can convert a weak optical signal into a larger electrical signal, so that the extremely weak change in the optical signal generated by the bubble reflection can be accurately detected, realizing high-sensitivity detection of the bubble.

[0050] In some embodiments, a lens 801 can be disposed between the excitation light emitter and the excitation light filter of the excitation light module 20, between the fluorescence detector and the fluorescence filter of the fluorescence detection module 30, or on one or both sides of the side where the optical element is disposed in the droplet detection channel 10, and between the first dichroic mirror 71 and the bubble detector 41, as shown in Figure 3 , Figure 4 , for converging the excitation light or fluorescence before entering the subsequent device.

[0051] The following uses specific embodiments to illustrate the setting modes of the droplet fluorescence detection device 1 including different numbers of excitation light modules 20 and fluorescence detection modules 30.

[0052] In some embodiments, the droplet fluorescence detection device 1 includes one excitation light module 20 and one fluorescence detection module 30. Specifically, as shown in Figures 3 - 4 :

[0053] The droplet fluorescence detection device 1 includes a first excitation light module 21, a first fluorescence detection module 31, a droplet detection channel 10, and a bubble detection module 40. The first excitation light module 21 includes a first excitation light emitter 211 and a first excitation light filter 212, as shown in Figure 3 . The first fluorescence detection module 31 includes a first fluorescence detector 311 and a first fluorescence filter 312, as shown in Figure 4 .

[0054] The bubble detection module 40 and the first fluorescence detection module 31 are disposed on the same side of the droplet detection channel 10. The droplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror 71.

[0055] Among them, which optical path the bubble detector 41 and the first fluorescence detector 311 are respectively disposed on is determined by the wavelength range detected by the detector, the cut-off wavelength of the first dichroic mirror 71, and the dichroic mirror type. Specifically, the maximum wavelength of the passband of the first excitation light filter 212 is less than the minimum wavelength of the passband of the first fluorescence filter 312, and the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312. Specifically, when the first dichroic mirror 71 is a long-pass dichroic mirror, the first fluorescence detector 311 and the first fluorescence filter 312 are both disposed on the transmission optical path of the first dichroic mirror 71, while the bubble detector 41 is disposed on the reflection optical path of the first dichroic mirror 71, as shown in Figure 4As shown. Specifically, when the first dichroic mirror 71 is a short-pass dichroic mirror, the first fluorescence detector 311 and the first fluorescence filter 312 are both arranged on the reflection optical path of the first dichroic mirror 71, and the bubble detector 41 is arranged on the transmission optical path of the first dichroic mirror 71.

[0056] Specifically, the first excitation light emitter 211 emits excitation light to irradiate the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter some of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The fluorescence passing through transmission or reflection is received by the first fluorescence detector 311, and the excitation light passing through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light filtered by the first excitation light filter 212.

[0057] In some embodiments, the first fluorescence detection module 31 is arranged on the first side of the droplet detection flow channel 10, and the bubble detection module 40 is arranged on the second side of the droplet detection flow channel 10, where the first side and the second side are different.

[0058] Among them, the first excitation light emitter 211 emits excitation light to irradiate the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter some of the excitation light. The emitted fluorescence is received by the first fluorescence detector 311, and the reflected and / or scattered excitation light is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light filtered by the first excitation light filter 212.

[0059] In some embodiments, the droplet fluorescence detection device 1 includes two excitation light modules 20 and two fluorescence detection modules 30. Specifically, see Figures 5 - 6 :

[0060] The droplet fluorescence detection device 1 includes a first excitation light module 21, a first fluorescence detection module 31, a second excitation light module 22, a second fluorescence detection module 32, a droplet detection flow channel 10, and a bubble detection module 40. The first excitation light module 21 includes a first excitation light emitter 211 and a first excitation light filter 212, and the second excitation light module 22 includes a second excitation light emitter 221 and a second excitation light filter 222. See Figure 5. The first fluorescence detection module 31 includes a first fluorescence detector 311 and a first fluorescence filter 312, and the second fluorescence detection module 32 includes a second fluorescence detector 321 and a second fluorescence filter 322, as shown in Figure 6 .

[0061] Among them, the maximum wavelength of the passband of the first excitation light filter 212 is less than the maximum wavelength of the passband of the second excitation light filter 222, and the maximum wavelength of the passband of the first fluorescence filter 312 is less than the maximum wavelength of the passband of the second fluorescence filter 322.

[0062] Among them, the bubble detection module 40 and the first fluorescence detection module 31 are arranged on the same side of the microdroplet detection flow channel 10. The microdroplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively arranged on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is arranged on the same side of the first fluorescence detection module 31 relative to the microdroplet detection flow channel 10. The microdroplet fluorescence detection device 1 further includes a second dichroic mirror 72. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively arranged on the transmission optical path or the reflection optical path of the second dichroic mirror 72.

[0063] In some embodiments, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0064] In some embodiments, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0065] Among them, which optical path the bubble detection module 40 and the first fluorescence detection module 31 are arranged on is determined by the wavelength range detected by the detector, the cut-off wavelength of the first dichroic mirror 71, and the type of dichroic mirror. In some embodiments, the first dichroic mirror 71 is a long-pass dichroic mirror. The first fluorescence detection module 31 is arranged on the transmission optical path of the first dichroic mirror 71, and the bubble detection module 40 is arranged on the reflection optical path of the first dichroic mirror 71, as Figure 6 shown. In some other embodiments, the first dichroic mirror 71 is a short-pass dichroic mirror. The first fluorescence detection module 31 is arranged on the reflection optical path of the first dichroic mirror 71, and the bubble detection module 40 is arranged on the transmission optical path of the first dichroic mirror 71.

[0066] Among them, which optical path the first fluorescence detection module 31 and the second fluorescence detection module 32 are set on is determined by the wavelength range detected by the detector, the cut-off wavelength of the second dichroic mirror 72, and the type of dichroic mirror. In some embodiments, the second dichroic mirror 72 is a long-pass dichroic mirror, the first fluorescence detection module 31 is set on the reflection optical path of the first dichroic mirror 71, and the second fluorescence detection module 32 is set on the transmission optical path of the second dichroic mirror 72. In other embodiments, the second dichroic mirror 72 is a short-pass dichroic mirror, the first fluorescence detection module 31 is set on the transmission optical path of the second dichroic mirror 72, and the second fluorescence detection module 32 is set on the reflection optical path of the second dichroic mirror 72.

[0067] Specifically, the first excitation light emitter 211 and the second excitation light emitter 221 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter part of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The part of the fluorescence that passes through transmission or reflection is received by the first fluorescence detector 311, the other part of the fluorescence that passes through transmission or reflection is received by the second fluorescence detector 321, and the excitation light that passes through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain fluorescent dyes that can be excited by the excitation light filtered by the first excitation light filter 212 and / or the second excitation light filter 222.

[0068] In some embodiments, the bubble detection module 40 and the first fluorescence detection module 31 are set on the same side of the droplet detection flow channel 10. The droplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively set on the transmission optical path or the reflection optical path of the first dichroic mirror 71; the second fluorescence detection module 32 is set on the opposite side of the first fluorescence detection module 31 with respect to the droplet detection flow channel 10. Of course, the first dichroic mirror 71 can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting method of the droplet fluorescence detection device 1 can be set according to its optical principle, which will not be elaborated here.

[0069] In some embodiments, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0070] In the embodiment of the present application, the first excitation light emitter 211 and the second excitation light emitter 221 emit excitation light that irradiates the droplet detection channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection channel 10 correspondingly reflect and / or scatter some of the excitation light. Part of the emitted fluorescence is received by the second fluorescence detector 321, and the other part of the emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The part of the fluorescence that has passed through transmission or reflection is received by the first fluorescence detector 311, and the excitation light that has passed through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light filtered by the first excitation light filter 212 and / or the second excitation light filter 222.

[0071] In some embodiments, the bubble detection module 40 is disposed on the first side of the droplet detection channel 10, and the first fluorescence detection module 31 and the second fluorescence detection module 32 are disposed on the second side of the droplet detection channel 10, and the first side is different from the second side; a second dichroic mirror 72 is further included between the first fluorescence detection module 31 and the second fluorescence detection module 32, and the first fluorescence detection module 31 and the second fluorescence detection module 32 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror. Among them, the second dichroic mirror 72 can be a long-pass dichroic mirror or a short-pass dichroic mirror, and the specific setting manners of the first fluorescence detection module 31 and the second fluorescence detection module 32 are correspondingly set according to the dichroic mirror type of the second dichroic mirror 72, which will not be elaborated here.

[0072] Specifically, the first excitation light emitter 211 and the second excitation light emitter 221 emit excitation light that irradiates the droplet detection channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection channel 10 correspondingly reflect and / or scatter some of the excitation light. The emitted fluorescence is received by the first fluorescence detector 311 and the second fluorescence detector 321, and the reflected and / or scattered excitation light is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by the excitation light filtered by the first excitation light filter 212 and / or the second excitation light filter 222.

[0073] In some embodiments, the droplet fluorescence detection device 1 includes three excitation light modules 20 and three fluorescence detection modules 30. Specifically, see Figures 7 - 8 :

[0074] The droplet fluorescence detection device 1 includes a first excitation light module 21, a first fluorescence detection module 31, a second excitation light module 22, a second fluorescence detection module 32, a third excitation light module 23, a third fluorescence detection module 33, a droplet detection flow channel 10, and a bubble detection module 40. 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, see Figure 7 . 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, see Figure 8 .

[0075] Specifically, the maximum wavelength of the passband of the first excitation light filter 212 is less than the maximum wavelength of the passband of the second excitation light filter 222 which is less than the maximum wavelength of the passband of the third excitation light filter 232, and the maximum wavelength of the passband of the first fluorescence filter 312 is less than the maximum wavelength of the passband of the second fluorescence filter 322 which is less than the maximum wavelength of the passband of the third fluorescence filter 332.

[0076] In some embodiments, the bubble detection module 40 is disposed between the droplet detection channel 10 and the first fluorescence detection module 31. The droplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is disposed on the same side of the first fluorescence detection module 31 as the droplet detection channel 10. A second dichroic mirror 72 is further disposed between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror 72. The third fluorescence detection module 33 is disposed on the same side of the first fluorescence detection module 31 and the second fluorescence detection module 32 as the droplet detection channel 10. A third dichroic mirror is further disposed between the second fluorescence filter 322 and the third fluorescence filter 332. The second fluorescence detector 321 and the third fluorescence detector 331 are respectively disposed on the transmission optical path or the reflection optical path of the third dichroic mirror. Wherein, the first dichroic mirror 71, the second dichroic mirror 72 and the third dichroic mirror can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting manners of the bubble detection module 40, the first fluorescence detection module 31, the second fluorescence detection module 32 and the third fluorescence detection module 33 are correspondingly set according to the dichroic mirror types of the first dichroic mirror 71, the second dichroic mirror 72 and the third dichroic mirror, which will not be elaborated herein.

[0077] Specifically, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0078] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0079] Specifically, the cut-off wavelength of the third dichroic mirror is greater than the maximum wavelength of the passband of the third fluorescence filter 332 and less than the minimum wavelength of the passband of the second fluorescence filter 322.

[0080] Specifically, the first excitation light emitter 211, the second excitation light emitter 221, and the third excitation light emitter 231 emit excitation light that irradiates the droplet detection channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection channel 10 correspondingly reflect and / or scatter part of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The fluorescence passing through transmission or reflection is respectively received by the first fluorescence detector 311, the second fluorescence detector 321, and the third fluorescence detector 331, and the excitation light passing through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by at least one excitation light filtered by the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232.

[0081] In some embodiments, the bubble detection module 40 and the first fluorescence detection module 31 are disposed on the same side of the droplet detection channel 10. The droplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is disposed on the same side of the first fluorescence detection module 31 relative to the droplet detection channel 10. A second dichroic mirror 72 is further disposed between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror 72; the third fluorescence detection module 33 is disposed on the opposite side of the first fluorescence detection module 31 and the second fluorescence detection module 32 relative to the droplet detection channel 10. Among them, the first dichroic mirror 71 and the second dichroic mirror 72 can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting manners of the bubble detection module 40, the first fluorescence detection module 31, and the second fluorescence detection module 32 are correspondingly set according to the dichroic mirror types of the first dichroic mirror 71 and the second dichroic mirror 72, which will not be elaborated here. Exemplarily, see Figure 8 , Figure 8 in which the first fluorescence detection module 31 and the second fluorescence detection module 32 are disposed on the same side of the droplet detection channel 10, and the third fluorescence detection module 33 is disposed on the opposite side of the first fluorescence detection module 31 and the second fluorescence detection module 32 relative to the droplet detection channel 10. Exemplarily, the first dichroic mirror 71 and the second dichroic mirror 72 are long-pass dichroic mirrors.

[0082] Specifically, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0083] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0084] In the embodiment of the present application, the first excitation light emitter 211, the second excitation light emitter 221, and the third excitation light emitter 231 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 emit fluorescence correspondingly, or the bubbles flowing through the droplet detection flow channel 10 reflect and / or scatter some of the excitation light. The partial fluorescence emitted is received by the third fluorescence detector 331. The partial fluorescence emitted or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The fluorescence passing through transmission or reflection is respectively received by the first fluorescence detector 311 and the second fluorescence detector 321, and the excitation light passing through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by at least one of the excitation lights filtered by the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232.

[0085] In some embodiments, the bubble detection module 40 is disposed on the first side of the droplet detection flow channel 10, and the first fluorescence detection module 31, the second fluorescence detection module 32, and the third fluorescence detection module 33 are disposed on the second side of the droplet detection flow channel 10, where the first side is different from the second side. Specifically, a second dichroic mirror 72 is further included between the first fluorescence detection module 31 and the first fluorescence detection module 31. The first fluorescence detection module 31 and the second fluorescence detection module 32 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror; a third dichroic mirror is further included between the second fluorescence detection module 32 and the third fluorescence detection module 33. The second fluorescence detection module 32 and the third fluorescence detection module 33 are respectively disposed on the transmission optical path or the reflection optical path of the third dichroic mirror. Among them, the second dichroic mirror 72 and the third dichroic mirror can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting manners of the first fluorescence detection module 31, the second fluorescence detection module 32, and the third fluorescence detection module 33 are set corresponding to the dichroic mirror types of the second dichroic mirror 72 and the third dichroic mirror, which will not be elaborated here.

[0086] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0087] Specifically, the cut-off wavelength of the third dichroic mirror is greater than the maximum wavelength of the passband of the third fluorescence filter 332 and less than the minimum wavelength of the passband of the second fluorescence filter 322.

[0088] Specifically, the first excitation light emitter 211, the second excitation light emitter 221, and the third excitation light emitter 231 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter part of the excitation light. The emitted fluorescence is received by the first fluorescence detector 311, the second fluorescence detector 321, and the third fluorescence detector 331, and the reflected and / or scattered excitation light is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain fluorescent dyes that can be excited by the excitation light filtered by the first excitation light filter 212, the second excitation light filter 222, and the third excitation light filter 232.

[0089] In some embodiments, the droplet fluorescence detection device 1 includes four excitation light modules 20 and four fluorescence detection modules 30. Specifically, see Figures 9 - 10 :

[0090] The droplet fluorescence detection device 1 includes a first excitation light module 21, a first fluorescence detection module 31, a second excitation light module 22, a second fluorescence detection module 32, a third excitation light module 23, a third fluorescence detection module 33, a fourth excitation light module 24, a fourth fluorescence detection module 34, a droplet detection flow channel 10, and a bubble detection module 40. 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. See Figure 9 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. See Figure 10 .

[0091] Specifically, the maximum wavelength of the passband of the first excitation light filter 212 is less than the maximum wavelength of the passband of the second excitation light filter 222, which is less than the maximum wavelength of the passband of the third excitation light filter 232, which is less than the maximum wavelength of the passband of the fourth excitation light filter 242. The maximum wavelength of the passband of the first fluorescence filter 312 is less than the maximum wavelength of the passband of the second fluorescence filter 322, which is less than the maximum wavelength of the passband of the third fluorescence filter 332, which is less than the maximum wavelength of the passband of the fourth fluorescence filter 342.

[0092] In some embodiments, the bubble detection module 40 and the first fluorescence detection module 31 are disposed on the same side of the microdroplet detection flow channel 10. The microdroplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is disposed on the same side of the first fluorescence detection module 31 with respect to the microdroplet detection flow channel 10. A second dichroic mirror 72 is further disposed between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror 72. The third fluorescence detection module 33 is disposed on the same side of the first fluorescence detection module 31 and the second fluorescence detection module 32 with respect to the microdroplet detection flow channel 10. A third dichroic mirror is further disposed between the second fluorescence filter 322 and the third fluorescence filter 332. The second fluorescence detector 321 and the third fluorescence detector 331 are respectively disposed on the transmission optical path or the reflection optical path of the third dichroic mirror. The fourth fluorescence detection module 34 is disposed on the same side of the first fluorescence detection module 31, the second fluorescence detection module 32, and the third fluorescence detection module 33 with respect to the microdroplet detection flow channel 10. A fourth dichroic mirror 74 is further disposed between the third fluorescence filter 332 and the fourth fluorescence filter 342. The third fluorescence detector 331 and the fourth fluorescence detector 341 are respectively disposed on the transmission optical path or the reflection optical path of the third dichroic mirror. Among them, the first dichroic mirror 71, the second dichroic mirror 72, the third dichroic mirror, and the fourth dichroic mirror 74 can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting manners of the bubble detection module 40, 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 correspondingly set according to the dichroic mirror types of the first dichroic mirror 71, the second dichroic mirror 72, the third dichroic mirror, and the fourth dichroic mirror 74, which will not be elaborated herein.

[0093] Specifically, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0094] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0095] Specifically, the cut-off wavelength of the third dichroic mirror is greater than the maximum wavelength of the passband of the third fluorescence filter 332 and less than the minimum wavelength of the passband of the second fluorescence filter 322.

[0096] Specifically, the cut-off wavelength of the fourth dichroic mirror 74 is greater than the maximum wavelength of the passband of the fourth fluorescence filter 342 and less than the minimum wavelength of the passband of the third fluorescence filter 332.

[0097] In the embodiment of the present application, 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 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 emit fluorescence correspondingly, or the bubbles flowing through the droplet detection flow channel 10 reflect and / or scatter part of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The fluorescence passing through transmission or reflection is respectively received by the first fluorescence detector 311, the second fluorescence detector 321, the third fluorescence detector 331, and the fourth fluorescence detector 341. The excitation light passing through transmission or reflection is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by at least one of the excitation lights 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.

[0098] In some embodiments, the bubble detection module 40 and the first fluorescence detection module 31 are arranged on the same side of the microdroplet detection flow channel 10. The microdroplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively arranged on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is arranged on the same side of the first fluorescence detection module 31 relative to the microdroplet detection flow channel 10. A second dichroic mirror 72 is further arranged between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively arranged on the transmission optical path or the reflection optical path of the second dichroic mirror 72. The third fluorescence detection module 33 is arranged on the same side of the first fluorescence detection module 31 and the second fluorescence detection module 32 relative to the microdroplet detection flow channel 10. A third dichroic mirror is further arranged between the second fluorescence filter 322 and the third fluorescence filter 332. The second fluorescence detector 321 and the third fluorescence detector 331 are respectively arranged on the transmission optical path or the reflection optical path of the third dichroic mirror. The fourth fluorescence detection module 34 is arranged on the side of the microdroplet detection flow channel 10 different from that of the first fluorescence detection module 31, the second fluorescence detection module 32 and the second fluorescence detection module 32. Among them, the first dichroic mirror 71, the second dichroic mirror 72 and the third dichroic mirror can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific arrangement manners of the bubble detection module 40, the first fluorescence detection module 31, the second fluorescence detection module 32 and the third fluorescence detection module 33 are arranged corresponding to the dichroic mirror types of the first dichroic mirror 71, the second dichroic mirror 72 and the third dichroic mirror, which will not be elaborated here.

[0099] Specifically, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0100] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0101] Specifically, the cut-off wavelength of the third dichroic mirror is greater than the maximum wavelength of the passband of the third fluorescence filter 332 and less than the minimum wavelength of the passband of the second fluorescence filter 322.

[0102] In the embodiment of the present application, 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 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter some of the excitation light. A part of the emitted fluorescence is received by the fourth fluorescence detector 341, and another part of the emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The transmitted or reflected fluorescence is respectively received by the first fluorescence detector 311, the second fluorescence detector 321, and the third fluorescence detector 331, and the transmitted or reflected excitation light is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected herein contain a fluorescent dye that can be excited by at least one of the excitation lights 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.

[0103] In some embodiments, the bubble detection module 40 and the first fluorescence detection module 31 are disposed on the same side of the droplet detection flow channel 10. The droplet fluorescence detection device 1 further includes a first dichroic mirror 71. The bubble detector 41 and the first fluorescence detector 311 are respectively disposed on the transmission optical path or the reflection optical path of the first dichroic mirror 71. The second fluorescence detection module 32 is disposed on the same side of the first fluorescence detection module 31 with respect to the droplet detection flow channel 10. A second dichroic mirror 72 is further disposed between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror 72. The third fluorescence detection module 33 is disposed on the opposite side of the first fluorescence detection module 31 and the second fluorescence detection module 32 with respect to the droplet detection flow channel 10. The fourth fluorescence detection module 34 is disposed on the same side of the third fluorescence detection module 33 with respect to the droplet detection flow channel 10. A fourth dichroic mirror 74 is further included between the third fluorescence detection module 33 and the fourth fluorescence detection module 34. The third fluorescence detector 331 and the fourth fluorescence detector 341 are respectively disposed on the transmission optical path or the reflection optical path of the fourth dichroic mirror 74. Among them, the first dichroic mirror 71, the second dichroic mirror 72, and the fourth dichroic mirror 74 can be long-pass dichroic mirrors or short-pass dichroic mirrors. The specific setting manners of the bubble detection module 40, 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 set corresponding to the dichroic mirror types of the first dichroic mirror 71, the second dichroic mirror 72, and the fourth dichroic mirror 74, which will not be elaborated here. Exemplarily, see Figure 10 , Figure 10 in which the first fluorescence detection module 31 and the second fluorescence detection module 32 are disposed on the same side of the droplet detection flow channel 10, the third fluorescence detection module 33 and the fourth fluorescence detection module 34 are disposed on the opposite side of the first fluorescence detection module 31 and the second fluorescence detection module 32 with respect to the droplet detection flow channel 10, and the first dichroic mirror 71, the second dichroic mirror 72, and the fourth dichroic mirror 74 are long-pass dichroic mirrors.

[0104] Specifically, the cut-off wavelength of the first dichroic mirror 71 is greater than the minimum wavelength of the passband of the first excitation light filter 212 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0105] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0106] Specifically, the cut-off wavelength of the fourth dichroic mirror 74 is greater than the maximum wavelength of the passband of the fourth fluorescence filter 342 and less than the minimum wavelength of the passband of the third fluorescence filter 332.

[0107] In an embodiment of the present application, 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 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 emit fluorescence correspondingly, or the bubbles flowing through the droplet detection flow channel 10 reflect and / or scatter some of the excitation light. Some of the emitted fluorescence is received by the third fluorescence detector 331 and the fourth fluorescence detector 341. Another part of the emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror 71. The first dichroic mirror 71 transmits one of the fluorescence or the excitation light, and reflects the other of the fluorescence or the excitation light. The fluorescence passing through or reflected is received by the first fluorescence detector 311 and the second fluorescence detector 321 respectively, and the excitation light passing through or reflected is received by the bubble detector 41. Wherein, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by at least one of the excitation lights 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.

[0108] In some embodiments, the bubble detection module 40 is disposed on the first side of the droplet detection flow channel 10, and 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 disposed on the second side of the droplet detection flow channel 10. A second dichroic mirror 72 is further disposed between the first fluorescence detection module 31 and the second fluorescence detection module 32. The first fluorescence detector 311 and the second fluorescence detector 321 are respectively disposed on the transmission optical path or the reflection optical path of the second dichroic mirror 72. A third dichroic mirror is further disposed between the second fluorescence detection module 32 and the third fluorescence detection module 33. The second fluorescence detector 321 and the third fluorescence detector 331 are respectively disposed on the transmission optical path or the reflection optical path of the third dichroic mirror. A fourth dichroic mirror 74 is further included between the third fluorescence detection module 33 and the fourth fluorescence detection module 34. The third fluorescence detector 331 and the fourth fluorescence detector 341 are respectively disposed on the transmission optical path or the reflection optical path of the fourth dichroic mirror 74. Wherein, the second dichroic mirror 72 and the fourth dichroic mirror 74 can be a long-pass dichroic mirror or a short-pass dichroic mirror. The specific setting manners 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 correspondingly set according to the dichroic mirror types of the second dichroic mirror 72 and the fourth dichroic mirror 74, which will not be elaborated here.

[0109] Specifically, the cut-off wavelength of the second dichroic mirror 72 is greater than the maximum wavelength of the passband of the second fluorescence filter 322 and less than the minimum wavelength of the passband of the first fluorescence filter 312.

[0110] Specifically, the cut-off wavelength of the third dichroic mirror is greater than the maximum wavelength of the passband of the third fluorescence filter 332 and less than the minimum wavelength of the passband of the second fluorescence filter 322.

[0111] Specifically, the cut-off wavelength of the fourth dichroic mirror 74 is greater than the maximum wavelength of the passband of the fourth fluorescence filter 342 and less than the minimum wavelength of the passband of the third fluorescence filter 332.

[0112] In the embodiments of the present application, 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 emit excitation light that irradiates the droplet detection flow channel 10. Under the irradiation of the excitation light, at least some of the droplets 11 to be detected flowing through the droplet detection flow channel 10 correspondingly emit fluorescence, or the bubbles flowing through the droplet detection flow channel 10 correspondingly reflect and / or scatter part of the excitation light. The emitted fluorescence is received by the first fluorescence detector 311, the second fluorescence detector 321, the third fluorescence detector 331, and the fourth fluorescence detector 341, and the reflected and / or scattered excitation light is received by the bubble detector 41. Among them, at least some of the droplets 11 to be detected here contain a fluorescent dye that can be excited by at least one of the excitation lights 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.

[0113] Those skilled in the art can understand that, in addition to the above embodiments, the droplet fluorescence detection device 1 of the present application may further include more than 4 excitation light modules 20, and the setting manner of the bubble detection module 40 and the fluorescence detection module 30 can refer to the above embodiments.

[0114] In some embodiments, the droplet fluorescence detection device 1 further includes a processor 50, which is configured to receive the fluorescence signals collected by the fluorescence detectors and the bubble signals collected by the bubble detector 41, determine the fluorescence signals synchronized with the bubble signals as invalid fluorescence signals, and after removing the invalid fluorescence signals from the fluorescence signals, determine the detection result of the fluid to be detected according to the fluorescence signals after removal.

[0115] Specifically, Figure 11 A schematic diagram showing the changes in fluorescence signals and bubble signals when only the droplets 11 to be detected flow through the droplet detection flow channel 10 is shown. Figure 12Shows a schematic diagram of the changes in fluorescence signals and bubble signals when a series of bubbles flow through the droplet detection channel 10. The horizontal axis represents time, and the vertical axis represents the signal amplitudes of fluorescence and excitation light detected by the fluorescence detector and the bubble detector 41.

[0116] Specifically, when the droplet 11 to be detected flows through the droplet detection channel 10, the excitation light excites the fluorescent dye to generate fluorescence. After the fluorescence detector receives the fluorescence, a fluorescence signal is generated, as shown in, for example, Figure 11 the waveform curve of the fluorescence signal in. For each droplet 11 to be detected passing through, a corresponding waveform of the fluorescence signal is generated. Since the bubble detector 41 is only sensitive to the part of the excitation light reflected and / or scattered by the bubble, when only the droplet 11 to be detected flows through the droplet detection channel 10, there is no signal in the bubble signal of the bubble detector 41, as shown in, for example, Figure 11 the waveform curve of the bubble signal in. Further, the processor 50 can retain this part of the fluorescence signal for the next operation to finally determine the detection result.

[0117] Specifically, when the bubble flows through the droplet detection channel 10 alone or simultaneously with the droplet 11 to be detected, after the excitation light is emitted or scattered by the bubble, it is received by the bubble detector to form a bubble signal, as shown in, for example, Figure 12 the waveform curve of the bubble signal in. Further, the processor 50 can eliminate the fluorescence signal synchronized with the bubble signal.

[0118] In some embodiments, the signal acquisition of the fluorescence detector and the bubble detector is synchronized. In other words, when the bubble and the droplet 11 to be detected flow through the droplet detection channel 10 simultaneously, the fluorescence received by the fluorescence detector and the excitation light received by the bubble detector are synchronized, as shown in, for example, Figure 12 . Since the bubble signal and the fluorescence signal are generated synchronously, the processor 50 can determine the fluorescence signal generated synchronously with the bubble signal as an invalid fluorescence signal, eliminate the invalid fluorescence signal, and determine the detection result of the fluid to be detected based on the fluorescence signal after elimination.

[0119] In some embodiments, the signal acquisition of the fluorescence detector and the bubble detector is not synchronized, but there is a preset time difference; for example, when the bubble and the droplet 11 to be detected flow through the droplet detection channel 10 simultaneously, there is a preset time difference between the fluorescence received by the fluorescence detector and the excitation light received by the bubble detector. Then, the processor 50 identifies the fluorescence signal generated synchronously with the bubble signal through a time correction method, determines it as an invalid fluorescence signal, eliminates the invalid fluorescence signal, and determines the detection result of the fluid to be detected based on the fluorescence signal after elimination.

[0120] 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 mutually exclusive with other embodiments. Those skilled in the art 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 each embodiment 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.

[0121] 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 this application has been described in detail with reference to the above preferred implementation manners, those of ordinary skill in the art should understand that the technical solutions of this application can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of this application.

[0122] 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 mutually exclusive with other embodiments. Those skilled in the art 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 each embodiment 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.

[0123] 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 this application has been described in detail with reference to the above preferred implementation manners, those of ordinary skill in the art should understand that the technical solutions of this application can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of this application.

Claims

1. A droplet fluorescence detection device, characterized in that: include: A droplet detection flow channel, for allowing a fluid to be detected to flow through, wherein the fluid to be detected includes at least droplets to be detected and bubbles; A first excitation light module, comprising a first excitation light emitter; A first fluorescence detection module, comprising a first fluorescence detector; A bubble detection module, including a bubble detector; a first dichroic mirror, wherein the bubble detector and the first fluorescence detector are respectively arranged on a transmission optical path or a reflection optical path of the first dichroic mirror; The first excitation light emitter emits excitation light to illuminate the droplet detection channel. Under the illumination of the excitation light, at least a portion of the droplets to be detected flowing through the droplet detection channel correspondingly emit fluorescence, or the bubbles flowing through the droplet detection channel correspondingly reflect and / or scatter a portion of the excitation light. The emitted fluorescence or the reflected and / or scattered excitation light passes through the first dichroic mirror, which transmits one of the fluorescence or the excitation light and reflects the other of the fluorescence or the excitation light. The transmitted or reflected fluorescence is received by the first fluorescence detector, and the transmitted or reflected excitation light is received by the bubble detector.

2. The droplet fluorescence detection device according to claim 1, characterized in that: The first excitation light module further includes a first excitation light filter, wherein the first excitation light emitter emits excitation light, which is filtered by the first excitation light filter and then irradiated onto the droplet detection channel; the first fluorescence detection module further includes a first fluorescence filter, wherein the fluorescence transmitted or reflected by the first dichroic mirror is filtered by the first fluorescence filter and then received by the first fluorescence detector; The first fluorescence detector and the first fluorescence filter are both arranged on the transmission optical path of the first dichroic mirror, and the bubble detector is arranged on the reflection optical path of the first dichroic mirror; The cut-off wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband of the first excitation light filter and smaller than the minimum wavelength of the passband of the first fluorescence filter.

3. The droplet fluorescence detection device according to claim 1, characterized in that: The first excitation light module further includes a first excitation light filter, wherein the first excitation light emitter emits excitation light, which is filtered by the first excitation light filter and then irradiated onto the droplet detection channel; the first fluorescence detection module further includes a first fluorescence filter, wherein the fluorescence transmitted or reflected by the first dichroic mirror is filtered by the first fluorescence filter and then received by the first fluorescence detector; The first fluorescence detector and the first fluorescence filter are both arranged on the reflective optical path of the first dichroic mirror, and the bubble detector is arranged on the transmissive optical path of the first dichroic mirror; The cut-off wavelength of the first dichroic mirror is greater than the minimum wavelength of the passband of the first excitation light filter and smaller than the minimum wavelength of the passband of the first fluorescence filter.

4. The microdroplet fluorescence detection device according to claim 1, characterized in that: The bubble detector includes a photodiode PD, an avalanche photodiode APD, a multi-pixel photon counter MPPC or a photomultiplier tube PMT.

5. The microdroplet fluorescence detection device according to claim 1, characterized in that: The first fluorescence detector receives the fluorescence synchronously with the bubble detector receiving the excitation light.

6. The microdroplet fluorescence detection device according to claim 1, characterized in that: It also includes at least one of a second excitation light module, a third excitation light module and a fourth excitation light module, and correspondingly includes at least one of a second fluorescence detection module, a third fluorescence detection module and a fourth fluorescence detection module; The second excitation light module includes a second excitation light emitter and a second excitation light filter, and the second fluorescence detection module includes a second fluorescence detector and a second fluorescence filter, wherein the second excitation light emitter emits excitation light, which passes through the second excitation light filter and then irradiates the droplet detection channel. Under the irradiation of the excitation light, at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence in response, and the fluorescence is filtered by the second fluorescence filter and then received by the second fluorescence detector. The third excitation light module includes a third excitation light emitter and a third excitation light filter, and the third fluorescence detection module includes a third fluorescence detector and a third fluorescence filter, wherein the third excitation light emitter emits excitation light, which passes through the third excitation light filter and then irradiates the droplet detection channel. Under the irradiation of the excitation light, at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, and the fluorescence is filtered by the third fluorescence filter and then received by the third fluorescence detector. The fourth excitation light module also includes a fourth excitation light emitter and a fourth excitation light filter, and the fourth fluorescence detection module includes a fourth fluorescence detector and a fourth fluorescence filter, wherein the fourth excitation light emitter emits excitation light, which, after passing through the fourth excitation light filter, is irradiated onto the droplet detection channel, and under the irradiation of the excitation light, at least part of the droplets to be detected flowing through the droplet detection channel emit fluorescence accordingly, and the fluorescence is filtered by the fourth fluorescence filter and then received by the fourth fluorescence detector.

7. The droplet fluorescence detection device according to claim 6, characterized in that: comprising the second excitation light module and the second fluorescence detection module; Also comprising a second dichroic mirror, the second dichroic mirror being disposed on a transmission optical path or a reflection optical path of the first dichroic mirror; The first fluorescence detector and the first fluorescence filter are arranged on one of the transmission optical path or the reflection optical path of the second dichroic mirror, and the second fluorescence detector and the second fluorescence filter are arranged on the other of the transmission optical path or the reflection optical path of the second dichroic mirror.

8. The droplet fluorescence detection device according to claim 7, characterized in that: The maximum wavelength of the first excitation light filter passband is smaller than the maximum wavelength of the second excitation light filter passband, and the maximum wavelength of the first fluorescence filter passband is smaller than the maximum wavelength of the second fluorescence filter passband.

9. The microdroplet fluorescence detection device according to claim 6, characterized in that: It includes a third excitation light module, a fourth excitation light module, a third fluorescence detection module and the fourth fluorescence detection module; Also comprising a third dichroic mirror, the third dichroic mirror being disposed on the transmission optical path or the reflection optical path of the first dichroic mirror; The third fluorescence detector and the third fluorescence filter are arranged on one of the transmission optical path or the reflection optical path of the third dichroic mirror, and the fourth fluorescence detector and the fourth fluorescence filter are arranged on the other of the transmission optical path or the reflection optical path of the third dichroic mirror; The maximum wavelength of the passband of the third excitation light filter is smaller than the maximum wavelength of the passband of the fourth excitation light filter, and the maximum wavelength of the passband of the third fluorescence filter is smaller than the maximum wavelength of the passband of the fourth fluorescence filter.

10. The droplet fluorescence detection device according to any one of claims 1 to 9, characterized in that: It also includes a processor, which is configured to receive the fluorescence signal collected by the first fluorescence detector and the bubble signal collected by the bubble detector, determine that the fluorescence signal synchronized with the bubble signal is an invalid fluorescence signal, remove the invalid fluorescence signal from the fluorescence signal, and determine the detection result of the fluid to be detected based on the fluorescence signal after removal.

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