Light path adjusting device for micro-channel fluorescence detection and detection equipment

Abstract

The invention discloses a light path adjusting device for micro-channel fluorescence detection and detection equipment, and relates to the technical field of measurement, the light path adjusting device comprises a main body, a dichroscope, a first polaroid and a second polaroid, a first light path channel and a second light path channel which are vertically crossed are arranged in the main body, the first light path channel is provided with a first end and a second end which are opposite, and the second light path channel is provided with a second end which is opposite to the first end; the second light path channel is provided with a third end and a fourth end which are opposite; the dichroscope is arranged at the intersection of the first light path channel and the second light path channel; the first polarizing film is arranged in the first light path channel and located between the dichroscope and the first end. The second polarizing film is arranged in the second light path channel and located between the dichroscope and the fourth end, and the polarization direction of the second polarizing film is perpendicular to the polarization direction of the first polarizing film. According to the light path adjusting device for micro-channel fluorescence detection and the detection equipment, a compact detection light path with wide applicability can be provided, the light path is easy to adjust, and the quality of collected light signals is better.

Description

Technical Field

[0001] The present invention relates to the field of measurement technology, and in particular to an optical path adjustment device and a detection equipment for microfluidic channel fluorescence detection. Background Art

[0002] Laser induced fluorescence is a measurement method commonly used in microfluidic chip analysis. During the process, the microfluidic chip needs to be observed and measured. For a microfluidic chip with multiple microchannels, multiple microchannels need to be scanned and measured. The microfluidic chip has multiple microchannels, and each microchannel must be connected to a guide tube to pass the object to be detected into the microchannel of the microfluidic chip. In order to complete the connection between the above-mentioned guide tube and the microchannel, one side surface of the microfluidic chip is usually selected as the connection surface for connecting the guide tube corresponding to the number of microchannels. The conventional measuring device is with the help of a fluorescence microscope, but due to the obstruction and interference of the external connection pipeline of the microfluidic chip, such a measuring device is not convenient for adjusting the optical path. For example, the surface of one side where the guide tube is connected to the microfluidic chip is the upper surface, and the surface on the opposite side is the lower surface. The optical path of the laser needs to be injected from the lower surface of the microfluidic chip, and the fluorescence microscope is set on the upper surface of the microfluidic chip to observe and measure the fluorescence in the microchannel. The above optical path structure is also difficult to perform independent fluorescence measurements on multiple microchannels in the microfluidic chip. Summary of the invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes an optical path adjustment device for microfluidic fluorescence detection, which can provide a compact and widely applicable detection optical path, and the optical path is easy to adjust and the quality of the collected optical signal is better.

[0004] The present invention also provides a detection device for microfluidic channel fluorescence detection.

[0005] According to the first aspect of the present invention, the optical path adjustment device for microfluidic fluorescence detection comprises a main body, wherein a first optical path channel and a second optical path channel are arranged vertically intersecting each other, wherein the first optical path channel has a first end and a second end opposite to each other, and the second optical path channel has a third end and a fourth end opposite to each other; A dichroic mirror, the dichroic mirror is arranged at the intersection of the first optical path channel and the second optical path channel, and the dichroic mirror can reflect the light incident from the first end to the third end; A first polarizer, the first polarizer is disposed in the first optical path, and the first polarizer is located between the dichroic mirror and the first end; A second polarizer, wherein the second polarizer is disposed in the second optical path channel, the second polarizer is located between the dichroic mirror and the fourth end, and the polarization direction of the second polarizer is perpendicular to the polarization direction of the first polarizer.

[0006] According to the optical path adjustment device for microchannel fluorescence detection of the embodiment of the present invention, there are at least the following beneficial effects: laser light can be injected from the first end of the first optical path channel, reflected by the dichroic mirror to be emitted toward the third end of the second optical path channel; the microfluidic chip can be placed on the end face of the third end of the main body, and the laser light emitted from the third end irradiates the microchannel of the microfluidic chip to excite fluorescence, and the fluorescence enters from the third end along the second optical path channel and is emitted toward the fourth end through the dichroic mirror; a light detector can be arranged at the fourth end to receive the fluorescence; in the above optical path structure, the laser irradiates the microfluidic chip and the fluorescence reflection The radiation occurs on the same side of the microfluidic chip, which makes it easy to adjust the position and distance between the microfluidic chip and the third end of the main body, that is, it is easy to adjust the detection optical path; before the laser is irradiated to the microfluidic chip, the laser with a specific polarization direction is filtered out by the first polarizer in the first optical path channel, and the fluorescence excited by the microfluidic chip irradiated by the laser is filtered out by the second polarizer in the second optical path channel. The polarization direction of the first polarizer is perpendicular to the polarization direction of the second polarizer. Even if part of the laser enters the second optical path channel, it will be filtered out by the second polarizer, which can improve the quality of the collected optical signal.

[0007] According to some embodiments of the present invention, the first optical path channel is provided with a blocking member, the blocking member is located between the dichroic mirror and the second end, and the blocking member is provided with a conical structure on a side facing the dichroic mirror, and the conical surface of the conical structure can reflect light.

[0008] According to some embodiments of the present invention, the inner wall of the first optical path channel is provided with an internal thread, the internal thread is located at the second end, and the blocking member is provided with an external thread, the external thread cooperates with the internal thread.

[0009] According to some embodiments of the present invention, the second optical path channel is provided with a plurality of lenses, the plurality of lenses are arranged along the light propagation direction of the second optical path channel, and the lenses are capable of gathering light.

[0010] According to some embodiments of the present invention, the second optical path channel is provided with a filter, and the filter is located between the dichroic mirror and the fourth end.

[0011] According to some embodiments of the present invention, the main body is connected to a switching component, the switching component is located on an end surface of the fourth end, and the switching component is used to connect to a light detector.

[0012] According to some embodiments of the present invention, the adapter assembly includes a sleeve and an adapter block, the sleeve is inserted into the second optical path channel and extends to the outside of the fourth end, the end face of the adapter block is provided with a plurality of penetrating connecting holes, the end face of the adapter block is also provided with a penetrating step hole, one end of the sleeve located outside the fourth end is inserted into the step hole, the adapter block is provided with a fixing hole, the fixing hole is located on the side of the step hole, and the fixing hole is connected to the step hole.

[0013] According to the second aspect of the present invention, the detection device for microchannel fluorescence detection is used in conjunction with a microfluidic chip having multiple microchannels. The detection device for microchannel fluorescence detection also includes the above-mentioned optical path adjustment device for microchannel fluorescence detection.

[0014] The detection device for microfluidic fluorescence detection according to the embodiment of the present invention has at least the following beneficial effects: the microfluidic chip can be placed on the end face of the third end of the optical path adjustment device for microfluidic fluorescence detection, and the laser irradiation of the microfluidic chip and the fluorescence excitation both occur on the same side of the microfluidic chip, which makes it easy to adjust the position and distance between the microfluidic chip and the third end of the main body, that is, it is easy to adjust the detection optical path; before the laser is irradiated to the microfluidic chip, the laser with a specific polarization direction is filtered out by the first polarizer in the first optical path channel, and the fluorescence excited by the microfluidic chip irradiated by the laser is filtered out by the second polarizer in the second optical path channel. The polarization direction of the first polarizer is perpendicular to the polarization direction of the second polarizer. Even if part of the laser enters the second optical path channel, it will be filtered out by the second polarizer, which can improve the quality of the collected optical signal.

[0015] According to some embodiments of the present invention, a clamping device is further included, wherein the clamping device includes a driving component and a slide, the slide is transmission-connected to the driving component, the slide is provided with at least two clamping arms, the clamping arms clamp the microfluidic chip on the slide, the optical path adjustment device for microchannel fluorescence detection is arranged below the slide, and a detection platform is provided on the end face of the third end, the driving component can drive the slide to move so that the surface of the microfluidic chip fits with the detection platform.

[0016] According to some embodiments of the present invention, multiple groups of independent flow-pushing devices are further included, each group of the flow-pushing devices has a flow guide tube, each of the flow guide tubes is connected to one of the microchannels, and the flow-pushing devices can push liquid into the microchannel.

[0017] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 A cross-sectional view of an optical path adjustment device for microfluidic channel fluorescence detection according to an embodiment of the present invention; Figure 2 An exploded view of an optical path adjustment device for microfluidic channel fluorescence detection according to an embodiment of the present invention; Figure 3 It is a schematic diagram of a partially enlarged structure of an optical path adjustment device for microfluidic channel fluorescence detection according to an embodiment of the present invention; Figure 4 It is a schematic diagram of the structure of a detection device for microfluidic channel fluorescence detection according to an embodiment of the present invention; Figure 5 is a structural schematic diagram of a light source assembly according to an embodiment of the present invention; Figure 6 is a schematic structural diagram of a driving assembly according to an embodiment of the present invention; Figure 7 It is a structural schematic diagram of a flow-pushing device according to an embodiment of the present invention; Figure 8 Schematic diagram of the optical path of fluorescence in an embodiment of the present invention; Fig. 9 Schematic diagram of comparison between a microchannel coating enhanced substrate and a common substrate of a microfluidic chip in an embodiment of the present invention; Fig.10 A schematic diagram showing the comparison between enhanced substrate-excited fluorescence and normal / reference substrate-excited fluorescence in an embodiment of the present invention; Fig.11 This is a schematic diagram of a pulse signal obtained by detecting fluorescence in an embodiment of the present invention; Fig.12 This is a standard curve diagram drawn when detecting different concentrations of the cancer marker NSE in an embodiment of the present invention.

[0019] Figure Number: Microfluidic chip 001; Main body 100, first optical path channel 110, first end 111, second end 112, second optical path channel 120, third end 121, fourth end 122, dichroic mirror 200, first polarizer 300, second polarizer 400, blocking member 510, lens 520, filter 530, adapter assembly 600, sleeve 610, adapter block 620, connecting hole 621, step hole 622, fixing hole 623; The driving assembly 710, the first guide driving rail 711, the second guide driving rail 712, the slide 720, the clamping arm 721, the flow pushing device 800, the flow guiding tube 810, the horizontal push slide 820, the horizontal push slider 830, the holder 840, and the injection tube 850; Light source assembly 910 , excitation light source 911 , filter tube 912 , collimating light guide channel 913 , first binder 920 , second binder 930 , and nanoparticles 940 . DETAILED DESCRIPTION

[0020] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0021] In the description of the present invention, it should be understood that descriptions involving orientation, such as orientation or positional relationship indicated as up, down, etc., are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0022] In the description of the present invention, "a plurality" means more than two. If there is a description of "first" or "second", it is only used for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0023] In the description of the present invention, unless otherwise clearly defined, terms such as setting, installing, connecting, etc. should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in the present invention based on the specific content of the technical solution.

[0024] As described in the background technology, the microfluidic chip is generally also provided with a plurality of microchannels, and the plurality of microchannels are parallel to each other. The microfluidic chip needs to be connected with a guide tube, and each guide tube is connected with a microchannel, and the liquid to be detected is injected into the microchannel. The principle of the laser induced fluorescence detection method using the microfluidic chip is that the sample to be detected is pushed into the microfluidic chip, and if there is a target in the sample, the target will be combined with the first binding substance embedded in advance in the microchannel, and then the reagent is pushed to the microfluidic chip, and there is a second binding substance with a fluorescent molecule label in the reagent, and if there is a detection target in the sample, the target has been captured by the first binding substance and fixed in the microchannel, and when the second binding substance flows through the microchannel, the second binding substance will also be combined with the target, so that the target carries a fluorescent molecule, and the microfluidic chip can be excited by laser irradiation. The brightness of the fluorescence can be detected using optical equipment such as a fluorescence microscope, and the concentration of the target in the sample can be judged by the brightness.

[0025] In the case of using a fluorescence microscope, the horizontal microfluidic chip is used as a reference, the lens of the fluorescence microscope is set on the upper surface of the microfluidic chip, and the laser is injected from the lower surface of the microfluidic chip to stimulate fluorescence. At the same time, the upper surface of the microfluidic chip is also connected to devices or components such as guide tubes. When it is necessary to move and adjust the optical path of the fluorescence entering the fluorescence microscope, the range of the adjustable optical path is limited by the restriction of the guide tube, and part of the laser will enter the fluorescence microscope to interfere with the optical signal.

[0026] Reference Figure 1 and Figure 2 As shown, an optical path adjustment device for microfluidic fluorescence detection according to an embodiment of the present invention includes a main body 100 , a dichroic mirror 200 , a first polarizer 300 and a second polarizer 400 .

[0027] A first optical path channel 110 and a second optical path channel 120 that cross each other vertically are provided in the main body 100. The first optical path channel 110 has a first end 111 and a second end 112 that are opposite to each other, and the second optical path channel 120 has a third end 121 and a fourth end 122 that are opposite to each other. The dichroic mirror 200 is arranged at the intersection of the first optical path channel 110 and the second optical path channel 120, and the dichroic mirror 200 can reflect the light incident from the first end 111 to the third end 121.

[0028] The first optical path channel 110 is used for the injection of laser. The light source assembly 910 capable of generating laser light can be docked with the first end 111 to inject the laser light into the first optical path channel 110. The laser light is irradiated from the first end 111 toward the direction of the second end 112. When passing through the dichroic mirror 200, it can be reflected by the dichroic mirror 200 to the second optical path channel 120 and emitted toward the third end 121. Therefore, the microfluidic chip 001 can be placed on the end face of the third end 121 of the main body 100, and the laser light emitted from the third end 121 can be irradiated on the microfluidic chip 001 to excite fluorescence. The fluorescence excited by the microfluidic chip 001 will be injected into the second optical path channel 120 and emitted from the third end 121 toward the fourth end 122. Therefore, an optical device such as a light detector can be set at the fourth end 122 to receive the fluorescence and detect it. In the above optical path structure, the microfluidic chip 001 is placed at the third end 121, the laser is emitted from the third end 121, and irradiates from the lower surface of the microfluidic chip 001, and the fluorescence excited by the microfluidic chip 001 is also emitted from the lower surface, and irradiates from the third end 121 toward the fourth end 122. When the detection optical path needs to be adjusted, it is only necessary to change the distance between the lower surface of the microfluidic chip 001 and the third end 121, and the guide tube is connected to the upper surface of the microfluidic chip 001, so it will not interfere with it, and it is easy to adjust the detection optical path.

[0029] The dichroic mirror 200 can almost completely transmit light of a specific wavelength and almost completely reflect light of other wavelengths, so a dichroic mirror 200 that can almost completely transmit fluorescence can be selected, and a laser with a suitable wavelength can be selected, and the laser can be almost completely reflected by the dichroic mirror 200. By adjusting the reflection angle of the dichroic mirror 200, the laser emitted from the first end 111 can be reflected to the third end 121. It should be understood that since the dichroic mirror 200 almost completely reflects the laser, some of the laser can still pass through the dichroic mirror 200 and be emitted from the second end 112.

[0030] The first polarizer 300 is disposed in the first optical path channel 110, and the first polarizer 300 is located between the dichroic mirror 200 and the first end 111; the second polarizer 400 is disposed in the second optical path channel 120, and the second polarizer 400 is located between the dichroic mirror 200 and the fourth end 122, and the polarization direction of the second polarizer 400 is perpendicular to the polarization direction of the first polarizer 300.

[0031] The laser propagates in the first optical path channel 110, and passes through the first polarizer 300 before being reflected by the dichroic mirror 200. The first polarizer 300 is used to filter out laser light with a specific polarization direction. For example, in some embodiments, after the laser light passes through the first polarizer 300, a laser light polarized in the horizontal plane is obtained. After the laser light irradiates the microfluidic chip 001 to excite fluorescence, it irradiates and propagates from the third end 121 toward the fourth end 122. Before the fluorescence enters the light detector, it first passes through the second polarizer 400. The second polarizer 400 is used to filter out fluorescence with a specific polarization direction, and the polarization direction of the second polarizer 400 is perpendicular to the polarization direction of the first polarizer 300. For example, after the fluorescence passes through the second polarizer 400, fluorescence polarized in the vertical plane is obtained. At the same time, the second polarizer 400 can also filter out laser light polarized in the horizontal plane. Specifically, even if part of the laser light enters the second optical path channel 120 from the first optical path channel 110, this part of the laser light is also the horizontally polarized laser light filtered out by the first polarizer 300. When passing through the second polarizer 400, due to the vertical polarization direction, this part of the laser light can be filtered out by the second polarizer 400. Therefore, with the cooperation of the first polarizer 300 and the second polarizer 400, the quality of the optical signal collected by the optical detector can be improved.

[0032] It is important to understand that referring to Figure 5 As shown, the light source assembly 910 includes an excitation light source 911, a filter tube 912 and a collimated light guide channel 913, the filter tube 912 is connected to the excitation light source 911, one end of the collimated light guide channel 913 is connected to the filter tube 912, and the other end of the collimated light guide channel 913 is connected to the first end 111 of the main body 100.

[0033] The excitation light source 911 is used to generate laser light, which is first filtered out by the filter tube 912 to further filter out stray light of other wavelengths, and then adjusted by the collimated light guide channel 913 to adjust the laser light into parallel light and guide it into the first optical path channel 110 from the first end 111. The filter tube 912 filters the laser light, and can further control the wavelength of the laser light within the required range. When the laser light irradiates the dichroic mirror 200, the reflectivity of the dichroic mirror 200 to the laser light can be increased, and the laser light passing through the dichroic mirror 200 can be reduced.

[0034] Since the dichroic mirror 200 does not completely reflect light of a specific wavelength, most of the laser light can be reflected by the dichroic mirror 200 to the second optical path channel 120, and a small portion of the laser light still passes through the dichroic mirror 200 and propagates toward the second end 112. Generally, the second end 112 is not blocked, and the laser light propagating toward the second end 112 can be emitted to the external environment. However, if the ambient light is bright enough, the ambient light can also enter the first optical path channel 110 through the second end 112, and the ambient light may also propagate toward the second optical path channel 120 after reflection, affecting the collection of optical signals.

[0035] It is understandable that the first optical path channel 110 is provided with a blocking member 510, and the blocking member 510 is located between the dichroic mirror 200 and the second end 112. The blocking member 510 is provided with a conical structure on the side facing the dichroic mirror 200, and the conical surface of the conical structure can reflect light.

[0036] The blocking member 510, on the one hand, blocks ambient light from entering the first optical path channel 110, and on the other hand, reduces the interference of the laser light passing through the dichroic mirror 200 on the optical signal. The blocking member 510 is provided with a cone structure on the side facing the dichroic mirror 200, and the cone surface of the cone structure is not perpendicular to the axial direction of the first optical path channel 110. When the light is irradiated on the cone surface, it will not return to the original path, that is, the light parallel to the first optical path channel 110 will not propagate parallel to the first optical path channel 110 again after being reflected by the cone surface.

[0037] The laser light passing through the dichroic mirror 200 propagates toward the blocking member 510. The laser light is irradiated on the conical structure of the blocking member 510. After being reflected by the conical surface, the laser light deviates from the axial direction of the first optical path channel 110 and irradiates toward the side wall of the first optical path channel 110. The laser light is gradually absorbed during multiple reflections on the side wall of the first optical path channel 110. The residual laser light that was originally collimated and concentrated is diverged and weakened, greatly reducing the possibility of it entering the second optical path channel 120, thereby reducing the interference of the laser light on the optical signal.

[0038] It is understandable that the inner wall of the first optical path channel 110 is provided with an internal thread, the internal thread is located at the second end 112, and the blocking member 510 is provided with an external thread, and the external thread cooperates with the internal thread. By means of thread cooperation, the blocking member 510 is fixed at the second end 112 of the first optical path channel 110. Further, a hexagonal hole can be provided at one end of the blocking member 510 facing the external environment, and the blocking member 510 can be screwed with a hexagonal wrench to complete screwing the blocking member 510 into the first optical path channel 110.

[0039] It can be understood that the second optical path channel 120 is provided with a plurality of lenses 520 , and the plurality of lenses 520 are arranged along the light propagation direction of the second optical path channel 120 , and the lenses 520 can gather light.

[0040] The second optical path channel 120 is mainly used for the propagation of the fluorescence stimulated by the microfluidic chip 001. The lens 520 is used to gather light, which can gather the divergent fluorescence and guide the fluorescence to propagate toward the fourth end 122. Gathering the fluorescence can improve the signal-to-noise ratio of the optical signal to a certain extent. Furthermore, the number and position of the lens 520 can be set and adjusted according to the actual light propagation situation. The adjusted fluorescence propagation optical path is as follows: Figure 8 For example, a lens 520 may be disposed between the dichroic mirror 200 and the third end 121, and the lens 520 may be used to initially focus the fluorescence. At the same time, when the collimated laser is reflected from the dichroic mirror 200 to the third end 121, it will also pass through the lens 520, and the laser will also be focused when passing through the lens 520, so that the laser can be controlled to accurately enter a single microchannel.

[0041] It is understandable that the second optical path channel 120 is provided with a filter 530 , and the filter 530 is located between the dichroic mirror 200 and the fourth end 122 .

[0042] The filter 530 is used to filter the laser and other stray light entering the second optical path channel 120, so that the fluorescence can pass through the filter 530, further reducing the interference of stray light on the optical signal. It should be understood that multiple filters 530 can be provided, for example, two filters 530 are provided in the second optical path channel 120.

[0043] Reference Figure 2 As shown, it can be understood that the main body 100 is connected to a transfer assembly 600 , the transfer assembly 600 is located at the end surface of the fourth end 122 , and the transfer assembly 600 is used to connect the light detector.

[0044] The adapter assembly 600 is used to stably connect the light detector to the main body 100, specifically to connect the light detector to the end face of the fourth end 122, and the fluorescence propagates in the second optical path channel 120, propagates from the third end 121 toward the fourth end 122, and then enters the light detector.

[0045] Reference Figure 3 As shown, it can be understood that the adapter assembly 600 includes a sleeve 610 and an adapter block 620, the sleeve 610 is inserted into the second optical path channel 120, and the sleeve 610 extends to the outside of the fourth end 122, the end face of the adapter block 620 is provided with a plurality of penetrating connecting holes 621, the end face of the adapter block 620 is also provided with a penetrating step hole 622, the end of the sleeve 610 located outside the fourth end 122 is inserted into the step hole 622, the adapter block 620 is provided with a fixing hole 623, the fixing hole 623 is located on the side of the step hole 622, and the fixing hole 623 is connected to the step hole 622.

[0046] First, the adapter block 620 is separated from the sleeve 610. To connect the adapter block 620 to the photodetector, a bolt can be used to pass through the connecting hole 621 and screw into the connecting end of the photodetector. After the adapter block 620 is connected to the photodetector, the adapter block 620 is moved so that the sleeve 610 is inserted into the step hole 622. Finally, a bolt or a locating pin can be used to screw into or insert into the outer wall of the sleeve 610 from the fixing hole 623 to achieve the connection between the sleeve 610 and the adapter block 620. The central channel of the sleeve 610 constitutes a part of the second optical path channel 120. It should be understood that some optical devices can also be arranged in the sleeve 610, such as the second polarizer 400, the lens 520 and the filter 530.

[0047] Reference Figure 4 As shown, a detection device for microchannel fluorescence detection according to an embodiment of the present invention is used in conjunction with a microfluidic chip 001, and the microfluidic chip 001 has multiple microchannels. The detection device for microchannel fluorescence detection also includes the above-mentioned optical path adjustment device for microchannel fluorescence detection.

[0048] It can be understood that it also includes a clamping device, which includes a driving component 710 and a slide 720. The slide 720 is connected to the driving component 710 by transmission. The slide 720 is provided with at least two clamping arms 721. The clamping arms 721 clamp the microfluidic chip 001 on the slide 720. The optical path adjustment device for microchannel fluorescence detection is arranged below the slide 720, and the end face of the third end 121 is provided with a detection platform. The driving component 710 can drive the slide 720 to move so that the surface of the microfluidic chip 001 fits with the detection platform.

[0049] The microfluidic chip 001 has a plurality of microchannels, which are parallel to each other, and each microchannel can be tested in turn. For example, the driving component 710 drives the slide 720 to move so that the first microchannel on the microfluidic chip 001 is aligned with the laser emitted from the third end 121. After the detection of the first microchannel is completed, the driving component 710 continues to drive the slide 720 to move so that the second microchannel on the microfluidic chip 001 is aligned with the laser, and the detection of the second microchannel is completed. The above steps are repeated until all microchannel detections on the microfluidic chip 001 are completed.

[0050] It is important to understand that, further, reference Figure 6 As shown, the driving assembly 710 includes a first guide driving rail 711 and a second guide driving rail 712. The first guide driving rail 711 is horizontally arranged, and the second guide driving rail 712 is vertically arranged. The second guide driving rail 712 is transmission-connected to the first guide driving rail 711, and the slide 720 is transmission-connected to the second guide driving rail 712.

[0051] In the above structure, the first guide drive rail 711 can drive the slide 720 to move horizontally, that is, to adjust the alignment of a specific microchannel on the microfluidic chip 001 with the laser, and the second guide drive rail 712 can drive the slide 720 to move vertically, that is, to adjust the distance between the lower surface of the microfluidic chip 001 and the detection platform at the third end 121.

[0052] It is understandable that it also includes multiple groups of independent flow-pushing devices 800, each group of flow-pushing devices 800 has a flow guide tube 810, each flow guide tube 810 is connected to a microchannel, and the flow-pushing devices 800 can push liquid into the microchannel.

[0053] Further, see Figure 7 As shown, the flow pushing device 800 includes a horizontal push slide 820, a horizontal push slider 830, a holder 840 and an injection tube 850. The horizontal push slider 830 is movably arranged on the horizontal push slide 820, and can drive the horizontal push slider 830 to move on the horizontal push slide 820. The holder 840 is arranged at one end of the horizontal push slide 820. When the horizontal push slide 820 moves, the distance between the horizontal push slide 820 and the holder 840 can be changed. One end of the injection tube 850 is fixedly connected to the holder 840, and the movable end of the injection tube 850 is connected to the horizontal push slide 820, that is, when the horizontal push slide 820 moves, the liquid in the injection tube 850 can be squeezed out. The injection tube 850 is connected to the guide tube 810, that is, the liquid in the injection tube 850 can be squeezed into the guide tube 810 and transferred to the microchannel of the microfluidic chip 001.

[0054] It should be understood that, in order to further enhance the fluorescence, an enhancement substrate may be coated in the microchannel of the microfluidic chip 001, wherein the enhancement substrate includes nanoparticles 940 and the first binding substance 920, such as Fig. 9 The first binding substance 920 can bind to the target in the sample to be detected, and the second binding substance 930 is labeled with a fluorescent molecule and can also bind to the target in the sample. The binding of the second binding substance 930 to the target does not affect the binding of the first binding substance 920 to the target. The fluorescence generated by the enhanced substrate and the fluorescence generated by the ordinary substrate can be referred to Fig.10 The effect shown in Fig.10 In the present invention, the reference substrate is the common substrate.

[0055] When the nanoparticles 940 in the enhanced substrate are exposed to a laser with a matching vibration frequency, localized surface plasmon resonance (LSPR) will occur, which is manifested as the nanoparticles 940 having a strong absorption effect on light waves, so that the originally dispersed light energy is bounded in a local area around the particles. The electromagnetic field in this area is significantly enhanced, thereby more strongly exciting the surrounding fluorescent molecules and enhancing the fluorescence signal.

[0056] In some detection embodiments, the target present in the sample to be detected is the cancer marker CEA, the fluorescent molecule is FITC, the first conjugate 920 is a CEA antibody, and the second conjugate 930 is a FITC-labeled CEA antibody. The first conjugate 920 and the second conjugate 930 are a pair of paired antibodies that can be simultaneously bound to different points of the target CEA. The sample is pushed to the microfluidic chip 001 by the flow-pushing device 800, first combined with the CEA antibody (first conjugate 920), and then the cleaning solution is pushed to wash away the unbound components to reduce interference, and then the FITC-labeled CEA antibody (second conjugate 930) is pushed to the microfluidic chip 001, and the FITC-labeled CEA antibody is combined with the target, and finally the cleaning solution is pushed to the microfluidic chip 001 again to wash away the unbound parts. The microfluidic chip 001 is translated, and each microchannel sweeps across the end surface of the third end 121 of the main body 100. The fluorescent molecule FITC is excited by the laser, and the generated fluorescence is collected by the light detector to form independent pulse signals, which correspond to the concentration value of the fluorescent-labeled analyte in each microchannel.

[0057] In some embodiments, the detection target is the cancer marker CEA, the fluorescent molecule is FITC, and the number of microchannels is 4. When each microchannel scans the end surface of the third end 121, if there is a fluorescently labeled second binder 930, the system can detect 4 independent pulse signals obtained in each scanning cycle. If there is no second binder 930, the system will detect a flat baseline. The detection result is shown in the following example. Fig.11 shown.

[0058] In some embodiments, the detection target is the cancer marker NSE, and the concentrations of the target are 1 ng / ml, 3.3 ng / ml, 10 ng / ml, 33.3 ng / ml, 100 ng / ml and 1000 ng / ml, respectively. The target is placed in different microchannels, and the fluorescent marker molecule is FITC. The samples in each microchannel are detected in turn according to the above method, and the standard curve drawn from the obtained detection results is as follows: Fig.12 shown.

[0059] The embodiments of the present invention are described in detail above with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the purpose of the present invention.

Claims

1. An optical path adjustment device for microfluidic fluorescence detection, characterized in that: include: A main body (100), wherein a first light path channel (110) and a second light path channel (120) which are perpendicularly intersecting are provided in the main body (100), wherein the first light path channel (110) has an opposite first end (111) and a second end (112), and the second light path channel (120) has an opposite third end (121) and a fourth end (122); a dichroic mirror (200), the dichroic mirror (200) being arranged at the intersection of the first optical path channel (110) and the second optical path channel (120), the dichroic mirror (200) being capable of reflecting light incident from the first end (111) to the third end (121); a first polarizer (300), the first polarizer (300) being arranged in the first light path channel (110), the first polarizer (300) being located between the dichroic mirror (200) and the first end (111); A second polarizer (400), the second polarizer (400) is arranged in the second light path channel (120), the second polarizer (400) is located between the dichroic mirror (200) and the fourth end (122), and the polarization direction of the second polarizer (400) is perpendicular to the polarization direction of the first polarizer (300).

2. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 1, characterized in that: The first optical path channel (110) is provided with a blocking member (510), the blocking member (510) is located between the dichroic mirror (200) and the second end (112), and a conical structure is provided on a side of the blocking member (510) facing the dichroic mirror (200), the conical surface of the conical structure being capable of reflecting light.

3. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 2, characterized in that: The inner wall of the first optical path channel (110) is provided with an internal thread, the internal thread is located at the second end (112), and the blocking member (510) is provided with an external thread, the external thread cooperates with the internal thread.

4. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 1, characterized in that: The second optical path channel (120) is provided with a plurality of lenses (520), the plurality of lenses (520) being arranged along the light propagation direction of the second optical path channel (120), and the lenses (520) being capable of gathering light.

5. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 1, characterized in that: The second optical path channel (120) is provided with a filter (530), and the filter (530) is located between the dichroic mirror (200) and the fourth end (122).

6. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 1, characterized in that: The main body (100) is connected to a transfer assembly (600), the transfer assembly (600) is located on the end surface of the fourth end (122), and the transfer assembly (600) is used to connect to a light detector.

7. The optical path adjustment device for microfluidic channel fluorescence detection according to claim 6, characterized in that: The adapter assembly (600) comprises a sleeve (610) and an adapter block (620), wherein the sleeve (610) is inserted into the second optical path channel (120), and the sleeve (610) extends to the outside of the fourth end (122), and the end surface of the adapter block (620) is provided with a plurality of penetrating connection holes (621), and the end surface of the adapter block (620) is also provided with a penetrating step hole (622), and one end of the sleeve (610) located outside the fourth end (122) is inserted into the step hole (622), and the adapter block (620) is provided with a fixing hole (623), and the fixing hole (623) is located on the side of the step hole (622), and the fixing hole (623) is communicated with the step hole (622).

8. A detection device for microfluidic fluorescence detection, used in conjunction with a microfluidic chip (001), wherein the microfluidic chip (001) has a plurality of microfluidic channels, characterized in that: The invention comprises the optical path adjustment device for microfluidic channel fluorescence detection according to any one of claims 1 to 7.

9. The detection device for microfluidic channel fluorescence detection according to claim 8, characterized in that: The invention also comprises a clamping device, wherein the clamping device comprises a driving component (710) and a slide (720), wherein the slide (720) is connected to the driving component (710) by transmission, and the slide (720) is provided with at least two clamping arms (721), wherein the clamping arms (721) clamp the microfluidic chip (001) on the slide (720), and the optical path adjustment device for microchannel fluorescence detection is arranged below the slide (720), and a detection platform is arranged on the end surface of the third end (121), and the driving component (710) can drive the slide (720) to move so that the surface of the microfluidic chip (001) is in contact with the detection platform.

10. The detection device for microfluidic fluorescence detection according to claim 9, characterized in that: It also includes a plurality of independent flow-pushing devices (800), each group of the flow-pushing devices (800) having a flow guide tube (810), each of the flow guide tubes (810) being connected to one of the microchannels, and the flow-pushing devices (800) being capable of pushing liquid into the microchannel.

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