Multimodal protein sample analysis equipment integrating fluorescence imaging and transmitted spectrum detection

Abstract

The invention discloses a multi-modal protein sample analysis device integrating fluorescence imaging and transmitted spectrum detection, which is characterized in that a low-magnification detection light path assembly is used for performing low-magnification scanning imaging on the whole general picture of a protein sample; the high-magnification fluorescence detection light path assembly is used for performing high-magnification bright field scanning imaging and high-magnification fluorescence scanning imaging on a protein sample; the filtering light path assembly is used for cooperating with the high-magnification fluorescence detection light path assembly to perform high-magnification absorbance scanning imaging on a protein sample so as to realize transmission spectrum detection; the object carrying motion holder is used for carrying a protein sample, switching between the low-magnification detection light path assembly and the high-magnification fluorescence detection light path assembly and carrying out imaging focusing. Low-magnification scanning imaging, high-magnification fluorescence scanning imaging and absorbance quantitative detection can be completed in single-time protein sample loading, the problems of data splitting and tedious operation are solved, and the method is suitable for sample-change-free multi-mode detection of protein samples.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical detection equipment, and in particular to a multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection. Background Art

[0002] Fluorescence microscopy technology (such as confocal microscopy and total internal reflection fluorescence microscopy) is one of the core tools for protein research. Through fluorescent protein labeling or immunofluorescence probes, protein subcellular localization, dynamic tracking and interaction analysis can be achieved.

[0003] Spectrophotometric techniques (such as UV-visible spectrometer, microplate reader) can quantitatively analyze protein concentration, modification status and enzyme activity by measuring the sample absorbance (such as characteristic absorption of protein at 280nm, absorption of specific wavelength after reaction of colorimetric reagent).

[0004] Although existing fluorescence microscopy microscopes can locate protein distribution, they cannot synchronously detect absorbance changes, and samples need to be transferred to spectrophotometric equipment (such as an ELISA reader) for chemical colorimetric analysis, resulting in sample loss, contamination risks, and data mismatch in time and space. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, one purpose of the present invention is to propose a multimodal protein sample analysis device that integrates fluorescence imaging and transmission spectrum detection, which can achieve low-magnification rapid scanning imaging, high-magnification fluorescence imaging, and absorbance quantitative detection in a single sample loading, and is mainly suitable for protein detection scenarios, solving the technical bottleneck that traditional single-function equipment requires multiple sample changes and data is difficult to correlate in time and space.

[0006] A multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to an embodiment of the present invention comprises:

[0007] A low-magnification detection optical path component, which is used to perform low-magnification scanning imaging of the overall appearance of the protein sample;

[0008] A high-magnification fluorescence detection optical path component, wherein the high-magnification fluorescence detection optical path component is used to perform high-magnification bright field scanning imaging and high-magnification fluorescence scanning imaging on the protein sample;

[0009] A filter optical path component, wherein the filter optical path component is used to cooperate with the high-magnification fluorescence detection optical path component to perform high-magnification absorbance scanning imaging on the protein sample to achieve transmission spectrum detection;

[0010] The object-carrying motion platform is used to carry the protein sample, switch between the low-magnification detection optical path component and the high-magnification fluorescence detection optical path component, and is used for imaging focusing.

[0011] When it is necessary to perform low-magnification scanning imaging of the protein sample, the protein sample is switched to the low-magnification detection optical path component through the movement of the carrier motion platform itself, and the low-magnification detection optical path component is used to perform fast and efficient low-magnification scanning imaging of the overall overview of the protein sample. When it is necessary to perform high-magnification fluorescence scanning imaging or high-magnification absorbance scanning imaging of the protein sample, the protein sample is switched to the high-magnification fluorescence detection optical path component through the movement of the carrier motion platform itself. In this way, low-magnification scanning imaging, high-magnification fluorescence scanning imaging, and absorbance quantitative detection can be completed in a single protein sample loading. At the same time, the data of low-magnification scanning imaging, high-magnification bright field scanning imaging, high-magnification fluorescence scanning imaging, and absorbance quantitative detection can all be integrated in the host computer of the multimodal protein sample analysis device of the integrated fluorescence imaging and transmission spectrum detection of the embodiment of the present invention for processing, which solves the technical bottleneck that traditional single-function equipment needs to change samples multiple times and data is difficult to correlate in time and space.

[0012] According to the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection in an embodiment of the present invention, by integrating a low-magnification detection optical path component, a high-magnification fluorescence detection optical path component, a filtering optical path component and a carrier motion gimbal, low-magnification scanning imaging, high-magnification fluorescence scanning imaging, and absorbance quantitative detection can be completed in a single protein sample loading, thereby solving the problems of data segmentation and cumbersome operation, and is suitable for sample-free multimodal detection of protein samples.

[0013] In some embodiments, the low-magnification detection optical path assembly and the high-magnification added-measurement optical path assembly are located above the object motion pan-tilt platform and are spaced side by side in the horizontal direction. The low-magnification detection optical path assembly and the high-magnification detection optical path assembly are both arranged vertically and upright, and the filter optical path assembly is located below the object motion pan-tilt platform.

[0014] In some embodiments, the high-magnification fluorescence detection optical path assembly includes a high-precision industrial camera, a tube lens, a beam splitter module, a microscope objective lens, and a first parallel light source module located on one side of the beam splitter module, which are arranged in sequence from top to bottom; wherein the beam splitter module includes a first lens group without a filtering function and a second lens group with a filtering function, the first lens group is used to switch to the optical path of the high-magnification fluorescence detection optical path assembly during high-magnification bright field scanning imaging, and the second lens group is used to switch to the optical path of the high-magnification fluorescence detection optical path assembly during high-magnification fluorescence scanning imaging.

[0015] In some embodiments, the first parallel light source module includes an LED light source, a first achromatic doublet lens, and a second achromatic doublet lens arranged in sequence; the second achromatic doublet lens is located between the first achromatic doublet lens and the first lens group or the second lens group of the beam splitter module in the optical path switched to the high-magnification fluorescence detection optical path assembly.

[0016] In some embodiments, the first lens group includes a first dichroic mirror arranged in an inclined manner; when the first lens group is switched to the optical path of the high-magnification fluorescence detection optical path assembly, the first lens group is located directly below the tube lens and directly above the microscope objective lens, and the first dichroic mirror can reflect the parallel light emitted by the first parallel light source module and focus it through the microscope objective lens to the protein sample located below the microscope objective lens, and the reflected light emitted by the protein sample passes through the microscope objective lens, the first dichroic mirror, and the tube lens in sequence and is collected by the high-precision industrial camera, thereby achieving high-magnification bright field scanning imaging;

[0017] The second lens group includes a second dichroic mirror arranged in an inclined manner, and an excitation filter and an emission filter located on both sides of the second dichroic mirror; when the second lens group is switched to the light path of the high-magnification fluorescence detection light path assembly, the second lens group is located directly below the tube lens and directly above the microscope objective lens, the excitation filter is located between the first parallel light source module and the second dichroic mirror, the emission filter is located between the tube lens and the second dichroic mirror, and the excitation filter can filter the parallel light emitted by the first parallel light source module to obtain a specific The second dichroic mirror can reflect the excitation light of the specific wavelength emitted by the excitation filter and focus it to the protein sample below the microscope objective lens through the microscope objective lens, so as to excite the fluorescent protein in the protein sample to emit fluorescence. The fluorescence emitted by the protein sample passes through the microscope objective lens and the second dichroic mirror in sequence to obtain transmitted light. The transmitted light emitted by the second dichroic mirror is filtered out by the emission filter to remove the excitation light of the specific wavelength, and then passes through the tube lens and is collected by the high-precision industrial camera, thereby realizing high-magnification fluorescence scanning imaging.

[0018] In some embodiments, there are multiple groups of the second lens group, and the wavelengths of the specific wavelength excitation light obtained by the excitation filters of different groups of the second lens groups are different, and the wavelengths of the specific wavelength excitation light filtered out by the emission filters of different groups of the second lens groups are different.

[0019] In some embodiments, the spectrometer module includes a mounting portion and a driving device; the mounting portion is installed with the first lens group and the second lens group, and the driving device is connected to the mounting portion for driving the mounting portion to move so as to switch the first lens group or the second lens group to the optical path of the high-magnification fluorescence detection optical path assembly.

[0020] In some embodiments, the filtering optical path component includes a second parallel light source module, a plano-convex lens, a plano-concave lens and a bandpass filter arranged in sequence; wherein, the parallel light emitted by the second parallel light source module is converged by the plano-convex lens and the plano-concave lens, and then obtains light of a specific wavelength through the bandpass filter, the light of the specific wavelength is incident on the protein sample from below the protein sample to the upper side and is absorbed by the protein sample to obtain protein sample transmitted light, the protein sample transmitted light passes through the microscope objective lens and the tube lens and is collected by the high-precision industrial camera, thereby completing high-magnification absorbance scanning imaging.

[0021] In some embodiments, the filter optical path assembly further includes a reflector, which is arranged obliquely directly below the microscope objective lens so that the light of the specific wavelength emitted by the bandpass filter is incident on the protein sample from below the protein sample upward.

[0022] In some embodiments, the bandpass filter is replaceable.

[0023] In some embodiments, the low-magnification detection optical path component includes an industrial camera, a FA lens, and a ring light source arranged in sequence from top to bottom.

[0024] In some embodiments, the object motion platform includes an X-axis electric slide, a Y-axis electric slide, a Z-axis electric lifting platform and a microfluidic chip fixing platform; the X-axis electric slide is arranged on the Y-axis electric slide, the Z-axis electric lifting platform is arranged on the X-axis electric slide, and the microfluidic chip fixing platform is arranged on the Z-axis electric lifting platform.

[0025] In some embodiments, the microfluidic chip fixing platform includes a base and a mounting plate; the base is disposed on the Z-axis electric lifting platform, and the mounting plate is slidably disposed on the upper side of the base.

[0026] 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

[0027] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0028] Figure 1a A schematic diagram of a multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to the present invention;

[0029] Figure 1b It is a schematic diagram of a 3D effect of a multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to the present invention;

[0030] Figure 1c Another perspective stereoscopic effect schematic diagram of the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention;

[0031] Figure 2 A schematic diagram of a low-magnification detection optical path component of a multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to the present invention;

[0032] Figure 3a A schematic diagram of a high-magnification fluorescence detection optical path component in one position of the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention;

[0033] Figure 3b A schematic diagram of another aspect of a high-magnification fluorescence detection optical path component of the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention;

[0034] Figure 3c for Figure 3b A is an enlarged schematic diagram;

[0035] Figure 4a A schematic diagram of a filter optical path component in one position of a multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention;

[0036] Figure 4b A schematic diagram of another orientation of the filter optical path component of the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention;

[0037] Figure 5 Schematic diagram of the object-carrying motion platform of the multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection of the present invention.

[0038] Reference numerals:

[0039] A multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection; a low-magnification detection optical path component 1; an industrial camera 101; an FA lens 102; a ring light source 103; a high-magnification fluorescence detection optical path component 2; a high-precision industrial camera 201; a tube lens 202; a beam splitter module 203; a second lens group 2031; a second dichroic mirror 20311; an excitation filter 20312; an emission filter 20313; a second frame 20314; a mounting portion 2032; a microscope objective 204; a first parallel light Source module 205; LED light source 2051; first achromatic doublet lens 2052; second achromatic doublet lens 2053; filter optical path component 3; second parallel light source module 301; plano-convex lens 302; plano-concave lens 303; bandpass filter 304; reflector 305; object motion platform 4; X-axis electric slide 401; Y-axis electric slide 402; Z-axis electric lift 403; microfluidic chip fixing platform 404; base 4041; mounting plate 4042; locking piece 4043; equipment rack 5. DETAILED DESCRIPTION

[0040] 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.

[0041] Combine the following Figures 1a to 5 The multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection according to an embodiment of the present invention is described.

[0042] like Figures 1a to 5 As shown, the multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection according to an embodiment of the present invention comprises a low-magnification detection optical path component 1, a high-magnification fluorescence detection optical path component 2, a filter optical path component 3 and a carrier motion platform 4. The low-magnification detection optical path component 1, the high-magnification fluorescence detection optical path component 2, the filter optical path component 3 and the carrier motion platform 4 are all integrated and installed on the equipment frame 5.

[0043] The low-magnification detection optical path component 1 is used to perform low-magnification scanning imaging of the overall profile of the protein sample. The purpose of low-magnification scanning imaging is to be able to better understand the overall profile of the protein sample before the subsequent high-magnification fluorescence scanning imaging or high-magnification absorbance scanning imaging of the protein sample, so as to reasonably select the local detection points of the protein sample for high-magnification fluorescence scanning imaging or high-magnification absorbance scanning imaging. By setting the low-magnification detection optical path component 1, the overall profile of the protein sample can be quickly and efficiently scanned at low magnification.

[0044] The high-magnification fluorescence detection optical path component 2 is used to perform high-magnification bright field scanning imaging and high-magnification fluorescence scanning imaging on the protein sample. Before using the high-magnification fluorescence detection optical path component 2 for high-magnification fluorescence scanning imaging, it is necessary to first use the high-magnification fluorescence detection optical path component 2 for high-magnification bright field scanning imaging. During this process, imaging focus is required to make the high-magnification bright field scanning imaging clear, in order to ensure that the high-magnification fluorescence scanning imaging in the next step is clear, thereby improving the efficiency and quality of the high-magnification fluorescence detection optical path component 2 for high-magnification fluorescence scanning imaging of protein samples.

[0045] The filter optical path component 3 is used to cooperate with the high-magnification fluorescence detection optical path component 2 to perform high-magnification absorbance scanning imaging on the protein sample to achieve transmission spectrum detection. Among them, the filter optical path component 3 is used to obtain light of a specific wavelength, and the specific wavelength light is absorbed by the protein sample to obtain the protein sample transmitted light, and the high-magnification fluorescence detection optical path component 2 collects the protein sample transmitted light to complete the high-magnification absorbance scanning imaging.

[0046] The object motion platform 4 is used to carry protein samples, switch between low-magnification detection optical path components and high-magnification fluorescence detection optical path components, and is used for imaging focusing.

[0047] When it is necessary to perform low-magnification scanning imaging on the protein sample, the protein sample is switched to the low-magnification detection optical path component 1 by the self-movement of the carrier motion platform 4, and the overall overview of the protein sample is quickly and efficiently scanned at low magnification using the low-magnification detection optical path component 1. When it is necessary to perform high-magnification fluorescence scanning imaging or high-magnification absorbance scanning imaging on the protein sample, the protein sample is switched to the high-magnification fluorescence detection optical path component 2 by the self-movement of the carrier motion platform. In this way, low-magnification scanning imaging, high-magnification fluorescence scanning imaging, and absorbance quantitative detection can be completed in a single protein sample loading. At the same time, the data of low-magnification scanning imaging, high-magnification bright field scanning imaging, high-magnification fluorescence scanning imaging, and absorbance quantitative detection can all be integrated in the host computer of the multimodal protein sample analysis device 1000 for integrated fluorescence imaging and transmission spectrum detection in the embodiment of the present invention for processing, which solves the technical bottleneck that traditional single-function equipment needs to change samples multiple times and data is difficult to correlate in time and space.

[0048] According to the multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection in an embodiment of the present invention, by integrating a low-magnification detection optical path component 1, a high-magnification fluorescence detection optical path component 2, a filtering optical path component 3 and a carrier motion platform 4, low-magnification scanning imaging, high-magnification fluorescence scanning imaging and absorbance quantitative detection can be completed in a single protein sample loading, thus solving the problems of data fragmentation and cumbersome operation, and being suitable for sample-free multimodal detection of protein samples.

[0049] In some embodiments, Figure 1a to Figure 1c As shown, the low-magnification detection optical path component 1 and the high-magnification additional measurement optical path component are arranged side by side and spaced apart in the horizontal direction above the carrier motion platform 4, the low-magnification detection optical path component 1 and the high-magnification detection optical path component 2 are both arranged vertically and upright, and the filter optical path component 3 is located below the carrier motion platform 4. Therefore, the low-magnification detection optical path component 1, the high-magnification detection optical path component 2, the filter optical path component 3 and the carrier motion platform 4 are arranged reasonably, which is conducive to the carrier motion platform 4 switching between the low-magnification detection optical path component 1 and the high-magnification additional measurement optical path component, and is conducive to the multimodal protein sample analysis device 1000 of the embodiment of integrated fluorescence imaging and transmission spectrum detection to realize multimodal detection without sample replacement, complete low-magnification scanning imaging, high-magnification fluorescence scanning imaging, and high-magnification absorbance scanning imaging, and solve the problems of data segmentation and cumbersome operation in the prior art.

[0050] In some embodiments, Figure 1a to Figure 1c as well as Figure 3a to Figure 3c As shown, the high-magnification fluorescence detection optical path component 2 includes a high-precision industrial camera 201, a tube lens 202, a beam splitter module 203, a microscope objective lens 204, and a first parallel light source module 205 located on one side of the beam splitter module 203, which are arranged in sequence from top to bottom; wherein the beam splitter module 203 includes a first lens group without a filtering function (the structure of the first lens group can refer to the structure of the second lens group 2031) and a second lens group 2031 with a filtering function, the first lens group is used to switch to the optical path of the high-magnification fluorescence detection optical path component 2 during high-magnification bright field scanning imaging, and the second lens group 2031 is used to switch to the optical path of the high-magnification fluorescence detection optical path component 2 during high-magnification fluorescence scanning imaging.

[0051] When high-magnification bright field scanning imaging is required, the first lens group of the beam splitter module 203 is switched to directly below the tube lens 202, that is, directly above the microscope objective lens 204, and the protein sample is switched to directly below the microscope objective lens 204 through the movement of the object moving platform 4; the first parallel light source module 205 is turned on, and the parallel light emitted by the first parallel light source module 205 is reflected downward through the first lens group and reaches the protein sample through the microscope objective lens 204. The height position of the protein sample is adjusted by the object moving platform 4 so that the microscope objective lens 204 is focused on the sample, and the reflected light emitted by the protein sample passes through the microscope objective lens 204, the first lens group and the tube lens 202 and is collected by the high-precision industrial camera 201, thereby achieving high-magnification bright field scanning imaging.

[0052] After completing high-magnification bright field scanning imaging, the second lens group 2031 is switched to the position directly below the tube lens 202 , that is, directly above the microscope objective lens 204 , and the first parallel light source module 205 is turned on to achieve high-magnification fluorescence scanning imaging.

[0053] In some embodiments, the first parallel light source module 205 includes an LED light source 2051, a first achromatic doublet lens 2052, and a second achromatic doublet lens 2053, which are arranged in sequence; the second achromatic doublet lens 2053 is located between the first achromatic doublet lens 2052 and the first lens group or the second lens group 2031 of the beam splitter module 203 in the optical path switched to the high-magnification fluorescence detection optical path component 2. The first achromatic doublet lens 2052 and the second achromatic doublet lens 2053 are used to convert the light emitted by the LED light source 2051 into parallel light, and the achromatic design ensures that the parallelism of light of different wavelengths is consistent after collimation.

[0054] In some embodiments, the first lens group includes a first dichroic mirror arranged in an inclined manner (such as at an inclined angle of 45 degrees); when the first lens group is switched to the light path of the high-magnification fluorescence detection optical path assembly 2, the first lens group is located directly below the tube lens 202 and directly above the microscope objective lens 204, and the first dichroic mirror can reflect the parallel light emitted by the first parallel light source module 205 and focus the emitted light through the microscope objective lens 204 to the protein sample located below the microscope objective lens 204. The reflected light emitted by the protein sample passes through the microscope objective lens 204, the first dichroic mirror, and the tube lens 202 in sequence and is collected by the high-precision industrial camera 201, thereby achieving high-magnification bright field scanning imaging.

[0055] Specifically, the first lens group (refer to the structure of the second lens group 2031) includes a first frame and a first dichroic mirror, a first non-filtering mirror and a second non-filtering mirror fixed on the first frame, wherein the first non-filtering mirror and the second non-filtering mirror are ordinary lenses without filtering function, and the first non-filtering mirror is located between the first parallel light source module 205 and the first dichroic mirror when the first lens group is switched to the light path of the high-magnification fluorescence detection optical path component 2, and the second non-filtering mirror is located between the tube lens 202 and the first dichroic mirror when the first lens group is switched to the light path of the high-magnification fluorescence detection optical path component 2. It should be noted that the first non-filtering mirror and the second non-filtering mirror may not be provided. The second lens group 2031 includes a second dichroic mirror 20311 arranged in an inclined manner (e.g., with an inclined angle of 45 degrees), an excitation filter 20312 and an emission filter 20313 located on both sides of the second dichroic mirror 20311; when the second lens group 2031 is switched to the light path of the high-magnification fluorescence detection optical path assembly 2, the second lens group 2031 is located directly below the tube lens 202 and directly above the microscope objective lens 204, the excitation filter 20312 is located between the first parallel light source module 205 and the second dichroic mirror 20311, the emission filter 20313 is located between the tube lens 202 and the second dichroic mirror 20311, and the excitation filter 20312 can transmit the first The parallel light emitted by the parallel light source module 205 is filtered to obtain an excitation light of a specific wavelength. The second dichroic mirror 20311 can reflect the excitation light of a specific wavelength emitted by the excitation filter 20312 and focus it through the microscope objective 204 to the protein sample located below the microscope objective 204, so as to excite the fluorescent protein in the protein sample to emit fluorescence. The fluorescence emitted by the protein sample passes through the microscope objective 204 and the second dichroic mirror 20311 in sequence to obtain transmitted light. The transmitted light emitted by the second dichroic mirror 20311 is filtered out by the emission filter 20313 to remove the excitation light of a specific wavelength, and then passes through the tube lens 202 and is collected by the high-precision industrial camera 201, thereby realizing high-magnification fluorescence scanning imaging.

[0056] Specifically, the second lens group 2031 includes a second frame 20314 and a second dichroic mirror 20311, an excitation filter 20312 and an emission filter 20313 fixed on the second frame 20314; when the second lens group 2031 is switched to the optical path of the high-magnification fluorescence detection optical path assembly 2, the second lens group 2031 is located directly below the tube lens 202 and directly above the microscope objective 204, the excitation filter 20312 is located between the first parallel light source module 205 and the second dichroic mirror 20311, and the emission filter 20313 is located between the tube lens 202 and the second dichroic mirror 20311.

[0057] In some embodiments, there is one first lens group, and there are multiple second lens groups 2031. For example, there are five second lens groups 2031. The excitation filters 20312 of different second lens groups 2031 obtain different wavelengths of specific wavelength excitation light, and the emission filters 20313 of different second lens groups 2031 filter out different wavelengths of specific wavelength excitation light. In this way, the required second lens group 2031 can be selected according to actual needs for high-magnification fluorescence scanning imaging, and different fluorescent proteins can be detected. It should be noted that in a high-magnification fluorescence measurement, usually only one first lens group and one second lens group 2031 are used.

[0058] In some embodiments, the beam splitter module 203 includes a mounting portion 2032 and a driving device (not shown in the figure); the mounting portion 2032 is mounted with a first lens group and a second lens group 2031, and the driving device is connected to the mounting portion 2032, and is used to drive the mounting portion 2032 to move, so as to switch the first lens group or the second lens group 2031 to the optical path of the high-magnification fluorescence detection optical path assembly 2. Specifically, the first lens group and multiple different second lens groups 2031 are circumferentially spaced, and the driving device such as a motor drives the mounting portion 2032 to rotate, thereby realizing the switching of the first lens group and the second lens group 2031 in the optical path of the high-magnification fluorescence detection optical path assembly 2.

[0059] In some embodiments, Figure 1a to Figure 1c as well as Figure 4a to Figure 4b As shown, the filtering optical path component 3 includes a second parallel light source module 301, a plano-convex lens 302, a plano-concave lens 303 and a bandpass filter 304 arranged in sequence; wherein the second parallel light source module 301 can be an LED parallel light source, and the parallel light emitted by the second parallel light source module 301 is converged by the plano-convex lens 302 and the plano-concave lens 303, and then obtains a specific wavelength of light through the bandpass filter 304, and the specific wavelength of light is incident on the protein sample from the bottom of the protein sample to the top and is absorbed by the protein sample to obtain the protein sample transmitted light, and the protein sample transmitted light passes through the microscope objective 204 and the tube lens 202 and is collected by the high-precision industrial camera 201, thereby completing high-magnification absorbance scanning imaging.

[0060] In some embodiments, when the second parallel light source module 301, the plano-convex lens 302, the plano-concave lens 303 and the bandpass filter 304 are arranged in sequence in the horizontal direction, the filter optical path component 3 further includes a reflector 305, and the reflector 305 is arranged obliquely (such as 45 degrees obliquely) directly below the microscope objective 204, so that the specific wavelength light emitted by the bandpass filter 304 is incident on the protein sample from the bottom of the protein sample. In this embodiment, the layout of the filter optical path component 3 is reasonable.

[0061] In some embodiments, the bandpass filter 304 is replaceable. In this way, the bandpass filter 304 can be replaced with a bandpass filter 304 with a different bandpass to obtain different specific wavelengths of light.

[0062] In some embodiments, Figure 1a to Figure 2 As shown, the low-magnification detection optical path component 1 includes an industrial camera 101, a FA lens 102 (i.e., an industrial lens) and a ring light source 103 arranged in sequence from top to bottom; wherein, the FA lens 102 has a larger aperture and has lower requirements on the brightness of the light source, and is suitable for imaging the overall profile of the protein sample, and can quickly provide good image quality within the entire field of view; the ring light source 103 can be an LED ring light source 103 to ensure that the light shining on the protein sample is uniform.

[0063] When low-magnification scanning imaging is required, the protein sample is switched to the position directly below the annular light source 103 through the self-movement of the object motion platform 4; the annular light source 103 is turned on, and the FA lens 102 and the industrial camera 101 are used for scanning imaging, thereby achieving low-magnification scanning imaging.

[0064] In some embodiments, Figure 1a to Figure 1c and Figure 5 As shown, the object motion platform 4 includes an X-axis electric slide 401, a Y-axis electric slide 402, a Z-axis electric lift 403 and a microfluidic chip fixing platform 404; the Y-axis electric slide 402 is arranged on the equipment rack 5, the X-axis electric slide 401 is arranged on the Y-axis electric slide 402, the Z-axis electric lift 403 is arranged on the X-axis electric slide 401, and the microfluidic chip fixing platform 404 is arranged on the Z-axis electric lift 403. Among them, the microfluidic chip fixing platform 404 is used to fix the microfluidic chip, the microfluidic chip is used to inject protein samples, the X-axis electric slide 401 and the Y-axis electric slide 402 are used to switch the carried protein samples between the low-magnification detection optical path component 1 and the high-magnification fluorescence detection optical path component 2, and the Z-axis slide is used to drive the protein sample to rise and fall, so as to realize automatic imaging focus.

[0065] In some embodiments, the microfluidic chip fixing platform 404 includes a base 4041 and a mounting plate 4042; the base 4041 is disposed on the Z-axis electric lifting platform 403, and the mounting plate 4042 is slidably disposed on the upper side of the base 4041. Specifically, a slide groove is provided on the base 4041, and the mounting plate 4042 can slidably cooperate with the slide groove to facilitate the replacement of protein samples.

[0066] In some embodiments, the microfluidic chip fixing platform 404 further includes a locking member 4043, which is disposed on the locking base 4041 and is used to lock and release the mounting plate 4042. When the mounting plate 4042 is locked, the protein sample can be prevented from shifting, and when the mounting plate 4042 is locked, the protein sample can be easily replaced.

[0067] The following is a specific example to describe the workflow of the multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection according to an embodiment of the present invention, as follows:

[0068] Step 1: Load the protein sample into the microfluidic chip, fix the microfluidic chip to the mounting plate 4042, and mount the mounting plate 4042 on the base 4041 and fix the position. Turn on the power, connect the host computer to the multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection according to the embodiment of the present invention, and control the X-axis electric slide 401 and the Y-axis electric slide 402 to place the protein sample in the low-magnification detection optical path component 1.

[0069] Step 2: Turn on the annular light source 103, control the vertical movement of the Z-axis electric lifting platform 403 to achieve focusing, use the industrial camera 101 to perform low-magnification scanning imaging of the protein sample, and turn off the annular light source 103 after imaging is completed.

[0070] Step 3: The host computer controls the X-axis electric slide 401 and the Y-axis electric slide 402 to place the protein sample at the high-magnification detection optical path component, turns on the LED light source 2051, controls the rotation of the beam splitter module 20311, so that the first lens group is in the optical path of the high-magnification fluorescence detection optical path component 2, uses the high-precision industrial camera 201 for imaging, controls the vertical movement of the Z-axis electric lift 403 to achieve focusing, and completes high-magnification bright field scanning imaging.

[0071] Step 4: Control the rotation of the beam splitter module 203, and switch the second lens group 2031 corresponding to the fluorescent protein to be detected to the optical path of the high-magnification fluorescence detection optical path component 2; the fluorescent protein in the protein sample emits fluorescence which is collected by the microscope objective 204, transmitted by the second dichroic mirror 20311, and the excitation light of a specific wavelength band is filtered by the emission filter 20313, and focused to the high-precision industrial camera 201 for collection through the tube lens 202. After the scanning imaging is completed, turn off the LED light source 2051 to complete the high-magnification fluorescence scanning imaging.

[0072] Step 5: Control the rotation of the beam splitter module 203, control the absence of filters in the light path, turn on the LED parallel light source, control the vertical movement of the Z-axis electric lifting platform 403 to achieve focusing, and the protein sample transmitted light passes through the microscope objective 204 and the tube lens 202 and is collected by the high-precision industrial camera 201 to complete high-magnification absorbance scanning imaging, and turn off the LED parallel light source.

[0073] Step 6: After the multimodal detection is completed, remove the microfluidic chip and turn off the power.

[0074] It should be noted that in the multimodal protein sample analysis device 1000 integrating fluorescence imaging and transmission spectrum detection in the embodiment of the present invention, optionally, the microscope objective 204 is a flat-field achromatic dry objective with a magnification of 10x and a working distance of 10.6 mm; optionally, the excitation filter 20312 has a diameter of 25 mm, a thickness of 3.5 mm, and a substrate material of ultraviolet fused quartz; optionally, the first dichroic mirror and the second dichroic mirror 20311 both have a diameter of 25.4 mm, a thickness of 3.2 mm, and a substrate material of ultraviolet fused quartz; optionally, the emission filter 20313 has a diameter of 25 mm, a thickness of 3.5 mm, and a substrate material of ultraviolet fused quartz; optionally, the tube lens 202 has a diameter of 23 mm, a thickness of 6 mm, and a substrate material of UV fused quartz; Optionally, the first achromatic doublet lens 2052 has a diameter of 25.4 mm, a center thickness of 15 mm, and the optical element material is BAFN11 / N-SF6; Optionally, the second achromatic doublet lens 2053 has a diameter of 50.8 mm, a center thickness of 13.5 mm, and the optical element material is H-LAK10 / H-ZF7LA; Optionally, the plano-convex lens 302 has a diameter of 25.4 mm, a center thickness of 3.6 mm, and the optical element material is N-BK7; Optionally, the plano-concave lens 303 has a diameter of 12.7 mm, a center thickness of 3 mm, and the optical element material is N-BK7; Optionally, the bandpass filter 304 has a diameter of 25 mm, a thickness of 3.5 mm, and the substrate material is UV fused quartz.

[0075] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0076] Although the embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the claims and their equivalents.

Claims

1. A multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection, characterized in that: include: A low-magnification detection optical path component, which is used to perform low-magnification scanning imaging of the overall appearance of the protein sample; A high-magnification fluorescence detection optical path component, wherein the high-magnification fluorescence detection optical path component is used to perform high-magnification bright field scanning imaging and high-magnification fluorescence scanning imaging on the protein sample; A filter optical path component, wherein the filter optical path component is used to cooperate with the high-magnification fluorescence detection optical path component to perform high-magnification absorbance scanning imaging on the protein sample to achieve transmission spectrum detection; The object-carrying motion platform is used to carry the protein sample, switch between the low-magnification detection optical path component and the high-magnification fluorescence detection optical path component, and is used for imaging focusing.

2. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 1, characterized in that: The low-magnification detection optical path component and the high-magnification additional measurement optical path component are located above the object moving pan-tilt platform in a horizontal direction and are spaced side by side. The low-magnification detection optical path component and the high-magnification detection optical path component are both arranged vertically and upright, and the filter optical path component is located below the object moving pan-tilt platform.

3. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 2, characterized in that: The high-magnification fluorescence detection optical path component includes a high-precision industrial camera, a tube lens, a spectrometer module, a microscope objective lens, and a first parallel light source module located on one side of the spectrometer module, which are arranged in sequence from top to bottom; wherein the spectrometer module includes a first lens group without a filtering function and a second lens group with a filtering function, the first lens group is used to switch to the optical path of the high-magnification fluorescence detection optical path component during high-magnification bright field scanning imaging, and the second lens group is used to switch to the optical path of the high-magnification fluorescence detection optical path component during high-magnification fluorescence scanning imaging.

4. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 3, characterized in that: The first parallel light source module includes an LED light source, a first achromatic doublet lens and a second achromatic doublet lens arranged in sequence; the second achromatic doublet lens is located between the first achromatic doublet lens and the first lens group or the second lens group of the beam splitter module in the optical path switched to the high-magnification fluorescence detection optical path assembly.

5. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 3, characterized in that: The first lens group includes a first dichroic mirror arranged in an inclined manner; when the first lens group is switched to the light path of the high-magnification fluorescence detection light path assembly, the first lens group is located directly below the tube lens and directly above the microscope objective lens, the first dichroic mirror can reflect the parallel light emitted by the first parallel light source module and focus it through the microscope objective lens to the protein sample located below the microscope objective lens, and the reflected light emitted by the protein sample passes through the microscope objective lens, the first dichroic mirror, the tube lens in sequence and is collected by the high-precision industrial camera, thereby realizing high-magnification bright field scanning imaging; The second lens group includes a second dichroic mirror arranged in an inclined manner, and an excitation filter and an emission filter located on both sides of the second dichroic mirror; when the second lens group is switched to the light path of the high-magnification fluorescence detection light path assembly, the second lens group is located directly below the tube lens and directly above the microscope objective lens, the excitation filter is located between the first parallel light source module and the second dichroic mirror, the emission filter is located between the tube lens and the second dichroic mirror, and the excitation filter can filter the parallel light emitted by the first parallel light source module to obtain a specific The second dichroic mirror can reflect the excitation light of the specific wavelength emitted by the excitation filter and focus it to the protein sample below the microscope objective lens through the microscope objective lens, so as to excite the fluorescent protein in the protein sample to emit fluorescence. The fluorescence emitted by the protein sample passes through the microscope objective lens and the second dichroic mirror in sequence to obtain transmitted light. The transmitted light emitted by the second dichroic mirror is filtered out by the emission filter to remove the excitation light of the specific wavelength, and then passes through the tube lens and is collected by the high-precision industrial camera, thereby realizing high-magnification fluorescence scanning imaging.

6. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 5, characterized in that: There are multiple groups of the second lens groups, and the wavelengths of the specific wavelength excitation light obtained by the excitation filters of different groups of the second lens groups are different, and the wavelengths of the specific wavelength excitation light filtered out by the emission filters of different groups of the second lens groups are different.

7. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 3, characterized in that: The spectrometer module includes a mounting portion and a driving device; the mounting portion is mounted with the first lens group and the second lens group, and the driving device is connected to the mounting portion and is used to drive the mounting portion to move so as to switch the first lens group or the second lens group to the optical path of the high-magnification fluorescence detection optical path assembly.

8. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 3, characterized in that: The filtering optical path component includes a second parallel light source module, a plano-convex lens, a plano-concave lens and a bandpass filter arranged in sequence; wherein, the parallel light emitted by the second parallel light source module is converged by the plano-convex lens and the plano-concave lens, and then obtains light of a specific wavelength through the bandpass filter, the light of the specific wavelength is incident on the protein sample from below the protein sample to the upper side and is absorbed by the protein sample to obtain protein sample transmitted light, the protein sample transmitted light passes through the microscope objective lens and the tube lens and is collected by the high-precision industrial camera, thereby completing high-magnification absorbance scanning imaging.

9. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 8, characterized in that: The filter optical path component also includes a reflector, which is arranged obliquely just below the microscope objective lens so that the light of the specific wavelength emitted by the bandpass filter is incident on the protein sample from below the protein sample upward.

10. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 8, characterized in that: The bandpass filter is replaceable.

11. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to any one of claims 1 to 10, characterized in that: The low-magnification detection optical path component comprises an industrial camera, a FA lens and a ring light source which are arranged in sequence from top to bottom.

12. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to any one of claims 1 to 10, characterized in that: The object motion platform includes an X-axis electric slide, a Y-axis electric slide, a Z-axis electric lift and a microfluidic chip fixing platform; the X-axis electric slide is arranged on the Y-axis electric slide, the Z-axis electric lift is arranged on the X-axis electric slide, and the microfluidic chip fixing platform is arranged on the Z-axis electric lift.

13. The multimodal protein sample analysis device integrating fluorescence imaging and transmission spectrum detection according to claim 12, characterized in that: The microfluidic chip fixing platform includes a base and a mounting plate; the base is arranged on the Z-axis electric lifting platform, and the mounting plate is slidably arranged on the upper side of the base.

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