Bioparticle Analysis Methods

Abstract

To provide a biological particle analysis method.SOLUTION: A biological particle analysis method includes the steps of: fluorescence staining a liquid specimen through a staining process so as to give a fluorescent color to at least one target biological particle; accommodating the liquid specimen into a minute liquid droplet generator and obtaining a real-time image of the liquid specimen using a camera device; dropping a target minute liquid droplet containing the target biological particle onto a biochip using the minute liquid droplet generator according to the real-time image; removing the fluorescent color of the target biological particle in the target minute liquid droplet through a washing process; and fluorescence staining the target biological particle on the biochip a plurality of times through the staining process and the washing process so as to obtain a plurality of fluorescence images corresponding to a plurality of kinds of biological characteristics.SELECTED DRAWING: Figure 8

Description

[Technical field]

[0001] The present invention relates to a method for analyzing biological particles, and more particularly to a method for analyzing biological particles using multiple fluorescent staining. [Background technology]

[0002] Conventional methods for analyzing bioparticles require multiple processes that cannot be smoothly linked together, starting from a liquid sample containing bioparticles to obtaining the final biocharacteristics. More specifically, at each stage in the conventional analytical methods, there are multiple types of processes, making it even more difficult to smoothly link different processes.

[0003] Therefore, in view of the fact that the above-mentioned problems can be improved, the inventors conducted intensive research and applied scientific theory, and as a result, arrived at the present invention as a method that is rational in design and can effectively improve the above-mentioned problems. Summary of the Invention [Problem to be solved by the invention]

[0004] The present invention provides a method for analyzing biological particles that effectively improves upon the problems in conventional analytical methods. [Means for solving the problem]

[0005] In an embodiment of the present invention, a bioparticle analysis method is provided, which includes a staining step of fluorescently staining a liquid specimen containing a plurality of bioparticles in a fluorescent staining process to give a fluorescent color to at least one of the plurality of bioparticles to make it a target bioparticle, an analysis step of storing the fluorescently stained liquid specimen in a microdroplet generating device and obtaining a real-time image of the liquid specimen stored in the microdroplet generating device with a camera device, a capture step of dropping a target microdroplet containing at least one of the target bioparticles on a biochip using the real-time image, and capturing at least one of the target bioparticles contained in the target microdroplet on the biochip, a washing step of removing the fluorescent color of the at least one of the target bioparticles captured on the biochip, and a fluorescent dyeing step of fluorescently staining the target bioparticles captured on the biochip N times by the fluorescent staining process and the washing process, and recording N types of biological characterizations in a recording device. and a characterization expressing step of obtaining a plurality of fluorescent images corresponding to a characteristic expression, N being a positive integer between 2 and 50. Effect of the Invention

[0006] As described above, the bioparticle analysis method according to the embodiment of the present invention can achieve a concentration effect when the capture step is performed by the staining step and the analysis step. Furthermore, the bioparticle analysis method can effectively connect the capture step, the washing step, and the characteristic display step by the biochip that captures the target bioparticle, thereby obtaining multiple fluorescent images corresponding to multiple biological characteristic expressions from the liquid sample through the bioparticle analysis method, which is advantageous for evaluating and judging the target bioparticles. [Brief description of the drawings]

[0007] [Figure 1]1 is a flowchart of a biological particle analysis method according to a first embodiment of the present invention. [Diagram 2] 1 is a schematic three-dimensional view showing an identification system according to a first embodiment of the present invention. [Diagram 3] FIG. 2 is a schematic side view of the identification system during a staining process. [Figure 4] FIG. 4 is a schematic diagram of the fluorescent staining process shown in FIG. [Diagram 5] FIG. 2 is a schematic side view of the identification system during an analysis process. [Figure 6] FIG. 6 is a partially enlarged view of FIG. [Figure 7] 1 is a schematic diagram of a real-time image of the inside of a microdroplet generating device captured by a camera device of the identification system; [Figure 8] FIG. 2 is a schematic side view of the identification system during a capture process. [Figure 9] FIG. 9 is a partially enlarged view of FIG. 8. [Figure 10] FIG. 2 is a schematic diagram of capturing a target bioparticle with a biochip of the identification system. [Figure 11] The biochip of the identification system is a schematic representation of a target bioparticle in another configuration. [Figure 12] FIG. 2 is a schematic diagram of the micro-droplet generating device of the identification system generating waste micro-droplets; [Figure 13] FIG. 4 is a schematic side view of the identification system during a cleaning process. [Figure 14] FIG. 4 is a schematic side view of the identification system during a characteristic display process. [Figure 15] FIG. 2 is a partial cross-sectional view of the camera device and the microdroplet generator of the identification system. [Figure 16] FIG. 4 is a schematic side view of an identification system according to a second embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] In order to better understand the characteristics and technical contents of the present invention, reference is made to the following detailed description and drawings of the present invention, but the description and drawings provided are for reference and explanation only and do not limit the present invention.

[0009] The present invention will be described below in accordance with certain specific embodiments, and those skilled in the art will be able to understand the advantages and effects of the present invention based on the contents disclosed herein. The present invention can be implemented or applied in other different specific embodiments, and various modifications and changes can be made to each detail in the present specification based on different perspectives and applications without departing from the concept of the present invention. In addition, as described in advance, the accompanying drawings of the present invention are simple schematic illustrations and are not drawn based on actual size. The technical contents of the present invention will be described in more detail based on the following embodiments, but the disclosed contents do not limit the scope of protection of the present invention.

[0010] It should be understood that, although the present specification may use the terms "first", "second", "third" and the like to describe various elements or signals, these elements or signals are not limited by these terms. These terms are mainly used to distinguish one element from another element or one signal from another signal. In addition, the term "or" used in the present specification may include any one or more combinations of the associated listed items according to actual circumstances.

[0011] [First embodiment] 1 to 15 show a first embodiment of the present invention. As shown in Fig. 1 and Fig. 2, in this embodiment, a bioparticle analysis method S100 and an identification system 100 are provided. The identification system 100 is applied to the bioparticle analysis method S100, and the specific arrangement of the identification system 100 can be adjusted according to design requirements, so the present invention is not limited thereto. For example, in other embodiments not shown in the drawings of the present invention, the bioparticle analysis method S100 can also be implemented via a structure different from that of the identification system 100.

[0012] In this embodiment, the identification system 100 includes a multiple fluorescent staining device 1, a microdroplet generating device 21, a camera device 23 corresponding to the microdroplet generating device 21, and a biochip 22 for connecting the multiple fluorescent staining device 1 and the microdroplet generating device 21. Among them, the multiple fluorescent staining device 1 may include a staining device 11, a cleaning device 12 adjacent to the staining device 11, and a recording device 13 (e.g., a camera) located corresponding to the staining device 11, but the present invention is not limited thereto.

[0013] 1 and 3 to 14, in this embodiment, the bioparticle analysis method S100 includes a staining step S110, an analyzing step S120, a capturing step S130, a washing step S140, and a characteristic display step S150 in this order, thereby effectively realizing enrichment of target bioparticles 201a and various biological characterization expressions in a liquid specimen 200 including a plurality of bioparticles 201. That is, any method that cannot perform the plurality of steps in order is different from the bioparticle analysis method S100 according to this embodiment.

[0014] 1, 3 and 4, in the staining step S110, the liquid specimen 200 is fluorescently stained in a fluorescent staining process to give a fluorescent color corresponding to a biological characteristic to at least one of the plurality of bioparticles 201, thereby forming the target bioparticle 201a. That is, the plurality of bioparticles 201 are divided into the target bioparticle 201a and other non-target bioparticles 201b.

[0015] In this embodiment, the liquid sample 200 is contained in a sample container 26. Furthermore, the fluorescent staining process is performed on the liquid sample 200 in the sample container 26 by the staining device 11 of the multiple fluorescent staining device 1, but the present invention is not limited to this. For example, in another embodiment of the present invention not shown in the drawings, the liquid sample 200 in the sample container 26 may be stained by a method other than the staining device 11. In this manner, the fluorescently stained liquid sample 200 is contained in the sample container 26.

[0016] In addition, the liquid sample 200 may be a body fluid sample (e.g., blood, lymph, saliva, ascites, or urine) derived from an animal, and the target bioparticles 201a may be a specific type of cell, such as circulating tumor cells (CTC), fetal nucleated red blood cells (FNRBC), exosomes, viruses, or bacteria, but the present invention is not limited thereto. For example, in another embodiment of the present invention not shown in the drawings, the liquid sample 200 may be a liquid sample derived from a plant.

[0017] 1, 5 and 7, in the analysis step S120, the fluorescently stained liquid specimen 200 is contained in a microdroplet generating device 21, and an instantaneous image of the liquid specimen 200 in the microdroplet generating device 21 is obtained by the camera device 23. In this embodiment, the microdroplet generating device 21 acquires the liquid specimen 200 from the specimen container 26, and the instantaneous image is obtained by the camera device 23, thereby obtaining the instantaneous position of the fluorescent target bioparticle 201a.

[0018] In the capture step S130, as shown in Fig. 1, Fig. 8 to Fig. 12, the microdroplet generating device 21 drops a target microdroplet 202a including at least one of the target bioparticles 201a onto the biochip 22 based on the real-time image. That is, in this embodiment, the microdroplet 202 (pico-droplet) dropped from the liquid specimen 200 by the microdroplet generating device 21 is divided into the target microdroplet 202a and waste microdroplet 202b not including the target bioparticle 201a. It should be noted that in this embodiment, the droplet 202 including the non-target bioparticle 201b but not including the target microdroplet 202a is defined as the waste microdroplet 202b.

[0019] More specifically, it can be confirmed whether the microdroplets 202 dropped by the microdroplet generating device 21 contain at least one of the target bioparticles 201a in the real-time image. Thus, each of the target microdroplets 202a is accurately dropped onto the biochip 22 so as to capture at least one of the target bioparticles 201a in the target microdroplets 202a in the biochip 22, and the waste microdroplets 202b are dropped into a waste container 27, thereby effectively concentrating the target bioparticles 201a.

[0020] It should be noted that in this embodiment, the biochip 22 is manufactured by a semiconductor process, and the specific structure of the biochip 22 can be adjusted according to design needs. For ease of understanding, the following provides a structure of the biochip 22 that can achieve a good capture effect, but the present invention is not limited thereto.

[0021] 10 and 11, the biochip 22 is used to support the target microdroplets 202a, and the biochip 22 can capture at least one of the target bioparticles 201a contained in the target microdroplets 202a. The biochip 22 includes a bottom layer 221, a plurality of capture arms 222 connected to the bottom layer 221 and spaced apart from each other, and a surface modification layer 223 formed at the ends of the plurality of capture arms 222. In the capture step S130, the biochip 22 can capture at least one of the target bioparticles 201a with the plurality of capture arms 222 and the surface modification layer 223.

[0022] More specifically, preferably, the surface modification layer 223 is further limited to an arginylglycylaspartic acid (RGD) peptide layer, thereby capturing the target bioparticle 201a by chemical bonding, but the present invention is not limited thereto. As shown in Fig. 11, the biochip 22 has at least two capture arms 222 that swing elastically relative to the bottom layer 221 to sandwich at least one target bioparticle 201a, which is advantageous for capturing the target bioparticle 201a by a physical method.

[0023] 1, 2, and 13, in the washing step S140, the fluorescent color of at least one of the target bioparticles 201a captured on the biochip 22 is removed in a washing process. In this embodiment, the washing process is performed on at least one of the target bioparticles 201a captured on the biochip 22 by the washing device 12 of the multiple fluorescent staining device 1.

[0024] 1, 2 and 14, the characteristic display step S150 involves fluorescently dyeing at least one of the target bioparticles 201a captured on the biochip 22 N times through the fluorescent dyeing process and the washing process, and obtaining a plurality of fluorescent images corresponding to N types of biological characterization expressions in the recording device 13. That is, the fluorescent dyeing of at least one of the target bioparticles 201a each time can correspond to one type of biological characterization expression. In this embodiment, N is a positive integer from 2 to 50, and is preferably from 4 to 28, but the present invention is not limited thereto.

[0025] It should be noted that the biochip 22 can be moved between different devices (e.g., the staining device 11, the washing device 12 and the stage 25) by manual or automatic means (e.g., a mechanical arm), but the present invention is not limited thereto.

[0026] As described above, the bioparticle analysis method S100 according to the embodiment of the present invention can achieve a concentration effect when performing the capture step S130 by the staining step S110 and the analysis step S120. In addition, the bioparticle analysis method S100 can effectively connect the capture step S130, the washing step S140, and the characteristic display step S150 by the biochip 22 that captures the target bioparticle 201a, so that the bioparticle analysis method S100 can obtain a plurality of fluorescent images corresponding to a plurality of biological characteristic expressions from the liquid sample 200, which is advantageous for evaluation and judgment of the target bioparticle 201a. In addition, the bioparticle analysis method S100 can further superimpose a plurality of fluorescent images to obtain the biological characteristics of the target bioparticle 201a.

[0027] As shown in Figures 1, 2 and 15, the above is a description of the steps in this embodiment of the bioparticle analysis method S100. For further understanding, the bioparticle concentration device 2 will be described below. In this embodiment, the bioparticle concentration device 2 is combined with a multiple fluorescent staining device 1 and used as equipment for carrying out the bioparticle analysis method S100, but the present invention is not limited thereto. For example, in other embodiments of the present invention not shown in the drawings, the bioparticle concentration device 2 may be used alone (e.g., for sale) or in combination with other devices.

[0028] In this embodiment, the bioparticle concentration device 2 includes the microdroplet generation device 21, the biochip 22 and the camera device 23 whose positions correspond to the microdroplet generation device 21, a control device 24 electrically connected to the microdroplet generation device 21 and the camera device 23, a loading platform 25 capable of supporting the biochip 22 and installed relative to the microdroplet generation device 21, the specimen container 26 and the waste container 27 installed on the loading platform 25, and a pressure balance mechanism 28 connected to the microdroplet generation device 21, but the present invention is not limited to this.

[0029] For example, in other embodiments not shown in the drawings of the present invention, the type and number of internal mechanisms of the bioparticle concentration device 2 may be adjusted according to design needs, and the microdroplet generation device 21 may be used alone for application (e.g., for sale) or in combination with other mechanisms (e.g., the biochip 22).

[0030] The mechanisms and their connections of the bioparticle concentrating device 2 will be described in this order below. Some of the mechanisms of the bioparticle concentrating device 2 (e.g., the biochip 22) have already been described above, so they will not be described here again.

[0031] The microdroplet generating device 21 includes a container 211, a hollow needle 212 communicating with the container 211, a first piezoelectric (PZT) member 213 disposed in the container 211, and a second piezoelectric member 214 disposed in the hollow needle 212. The container 211 is used to contain the liquid sample 200. The container 211 transports the liquid sample 200 to the hollow needle 212, and the liquid sample 200 is contained in the hollow needle 212.

[0032] More specifically, the container 211 includes a bottom surface 2111 and an enclosing side wall 2112 connected to the bottom surface 2111. The hollow needle 212 includes a connection end 2121 and a free end 2122 which are opposite ends, and the hollow needle 212 communicates (vertically) with the bottom surface 2111 of the container 211 via the connection end 2121. It is preferable that the inner diameter D211 of the container 211 is 5 to 30 times the inner diameter D212 of the hollow needle 212, and the inner diameter D212 of the hollow needle 212 refers to a portion other than the free end 2122 and is 300 μm to 700 μm. It should be noted that in this embodiment, the inner diameter D2122 of the free end 2122 of the hollow needle 212 is 30 μm to 150 μm, but the present invention is not limited thereto.

[0033] The first piezoelectric member 213 is arranged in a ring shape on the surrounding sidewall 2112 of the container 211, and in this embodiment, the first piezoelectric member 213 may be called an upper piezoelectric member 213. The first piezoelectric member 213 vibrates the container 211 to move the plurality of bioparticles 201 in the container 211 away from the surrounding sidewall 2112, or the first piezoelectric member 213 vibrates the container 211 to align the plurality of bioparticles 201 in the container 211 along a predetermined path.

[0034] More specifically, the predetermined path is disposed along the central axis L (and the direction of gravity) of the hollow needle 212, and the first piezoelectric member 213 has a distance D213 of 0.2 cm to 2 cm between it and the connection end 2121 along a direction parallel to the direction of the central axis L. Note that 20% to 85% of the area of ​​the surrounding side wall 2112 of the container 211 is covered with the first piezoelectric member 213. Also, the first piezoelectric member 213 may use a single one-piece structure having an annular shape, or may use a multi-piece structure arranged in an annular shape, depending on design needs.

[0035] The second piezoelectric member 214 is disposed on the outer surface of the hollow needle 212, and in this embodiment, the second piezoelectric member 214 may be referred to as a bottom piezoelectric member 214. The second piezoelectric member 214 squeezes the hollow needle 212 to cause the liquid sample 200 to flow outward and pass through the free end 2122 to form the microdroplets 202. The microdroplets 202 formed by the microdroplet generating device 21 are defined as a target microdroplet 202a containing at least one of the target bioparticles 201a and a waste microdroplet 202b containing no target bioparticle 201a.

[0036] More specifically, the second piezoelectric member 214 has a distance D214 of 0.2 cm to 2 cm between it and the free end 2122 along a direction parallel to the direction of the central axis L. 20% to 85% of the area of ​​the outer surface of the hollow needle 212 is covered with the second piezoelectric member 214. Also, the second piezoelectric member 214 may use a single one-piece structure having an annular shape, or may use a multi-piece structure arranged in an annular shape, depending on design needs.

[0037] It should be noted that the pressure balance mechanism 28 communicates with the container 211 so as to maintain the liquid sample 200 in the container 211 and the hollow needle 212 at a predetermined pressure. In this embodiment, the pressure balance mechanism 28 is described based on the following structure, but the present invention is not limited thereto. The pressure balance mechanism 28 includes an air pump 281, a switch 282 connected to the air pump 281, a pressure balance bottle 283 connected to the air pump 281 and the switch 282, and a liquid injection bottle 284 connected to the switch 282 and the container 211.

[0038] The biochip 22, the specimen container 26 and the waste container 27 are all placed on the platform 25. The platform 25 and the microdroplet generating device 21 can move relative to each other (for example, a multi-axis moving mechanism is installed on the platform 25). As a result, the microdroplet generating device 21 (in the analysis step S120) sucks the liquid specimen 200 from the specimen container 26 at the free end 2122 of the hollow needle 212 by the relative movement with the platform 25 (for example, the liquid specimen 200 is sucked into the container 211 and the hollow needle 212). In addition, the microdroplet generating device 21 can drop the target microdroplet 202a onto the biochip 22 and drop the waste microdroplet 202b into the waste container 27 by the relative movement with the platform 25.

[0039] The position of the camera device 23 corresponds to the hollow needle 212, and the camera device 23 (in the analysis step S120) obtains an instantaneous image of the liquid sample 200 at the free end 2122. The control device 24 is electrically connected to the second piezoelectric member 214 and the camera device 23, and the control device 24 (in the capture step S130) drives the second piezoelectric member 214 when at least one of the target bioparticles 201a is at the free end 2122 according to the instantaneous image. Thereby, the second piezoelectric member 214 squeezes the hollow needle 212, causing the liquid sample 200 to flow outward and pass through the free end 2122 of the hollow needle 212 to form the target microdroplet 202a.

[0040] As described above, the bioparticle concentrating device 2 according to an embodiment of the present invention can prevent the multiple bioparticles 201 from adhering to the inner wall of the container 211 by vibration of the first piezoelectric member 213 (or the upper piezoelectric member 213), and also moves and concentrates the multiple bioparticles 201 to the center of the container 211, which is advantageous for the concentrating process of the target bioparticles 201a.

[0041] To further explain, the bioparticle analysis method S100 or bioparticle concentration device 2 according to the embodiment of the present invention employs a combination of the camera device 23 and the second piezoelectric member 214 (or bottom piezoelectric member 214) to drive the second piezoelectric member 214 to form the target microdroplet 202a when at least one target bioparticle 201a is at the free end 2122 by the real-time image of the liquid sample 200 at the free end 2122. In this way, the concentration process of the target bioparticle 201a is effectively realized.

[0042] [Second embodiment] Figure 16 shows a second embodiment of the present invention. This embodiment is similar to the first embodiment, so the similar parts of the two embodiments will not be described repeatedly. The differences between this embodiment and the first embodiment will be described below.

[0043] In this embodiment, a single piezoelectric member may be adopted in the microdroplet generation device 21. That is, in this embodiment, the microdroplet generation device 21 omits the container 211 and the first piezoelectric member 213 (or the upper piezoelectric member 213) of the first embodiment shown in Fig. 3. Thus, in this embodiment, the microdroplet generation device 21 includes the hollow needle 212 and the second piezoelectric member 214 (or the bottom piezoelectric member 214) disposed on the outer surface of the hollow needle 212.

[0044] To further explain, the connection end 2121 of the hollow needle 212 is connected to the pressure balance mechanism 28 (e.g., the liquid injection bottle 284) and accommodates the liquid sample 200 in the hollow needle 212. Thereby, in the capture step S130, the microdroplet generating device 21 squeezes the hollow needle 212 with the second piezoelectric member 214 (or the bottom piezoelectric member 214) to make the liquid sample (not shown) flow out and pass through the free end 2122 of the hollow needle 212 to form the target microdroplet 202a, but the present invention is not limited thereto. For example, in other embodiments not shown in the drawings of the present invention, the microdroplet generating device 21 may adopt a structure other than a piezoelectric structure according to design needs.

[0045] [Advantageous Effects of the Embodiments] As described above, the bioparticle analysis method according to the embodiment of the present invention can achieve a concentration effect when the capture step is performed by the staining step and the analysis step. Furthermore, the bioparticle analysis method can effectively connect the capture step, the washing step, and the characteristic display step by the biochip that captures the target bioparticle, thereby obtaining multiple fluorescent images corresponding to multiple biological characteristic expressions from the liquid sample through the bioparticle analysis method, which is advantageous for evaluating and judging the target bioparticles.

[0046] Furthermore, in the bioparticle concentrating device according to an embodiment of the present invention, the vibration of the first piezoelectric member (or the upper piezoelectric member) can prevent a plurality of the bioparticles from adhering to the inner wall of the container, and also moves and concentrates the bioparticles to the center of the container, which is advantageous for the concentrating process of the target bioparticles.

[0047] To further explain, the bioparticle analysis method or the bioparticle concentration device according to the embodiment of the present invention employs a combination of the camera device and the second piezoelectric member (or bottom piezoelectric member) to drive the second piezoelectric member to form the target microdroplet when at least one of the target bioparticles is at the free end by the real-time image of the liquid sample at the free end, thus effectively realizing the concentration process of the target bioparticles.

[0048] The above disclosure is merely a preferred embodiment of the present invention, and the scope of the claims of the present invention is not limited thereto. Therefore, all equivalent technical modifications made by utilizing the contents of the specification and drawings of the present invention are included in the scope of the claims of the present invention. [Explanation of symbols]

[0049] S100…Bioparticle Analysis Method S110...Dyeing process S120…Analysis process S130…Capturing process S140…Cleaning process S150…Characteristics display process 100…Identification system 1. Multiple fluorescent staining device 11...Dyeing equipment 12...Cleaning equipment 13...Recording device 2. Biological particle concentration device 21...Micro droplet generation device 211…Container 2111…Bottom 2112…Enclosure side wall 212…Hollow needle 2121…Connection end 2122...Free end 213...first piezoelectric member (or upper piezoelectric member) 214...second piezoelectric member (or bottom piezoelectric member) 22. Biochip 221…Bottom layer 222…Capture arm 223...Surface modification layer 23...Camera equipment 24...Control device 25…Loading platform 26...Sample container 27...Waste container 28...Pressure balance mechanism 281…Air pump 282…Switch 283…Pressure balance bottle 284...Liquid injection bottle 200…Liquid sample 201...Bioparticles 201a…Target bioparticle 201b…Non-targeted bioparticles 202...Minute droplet 202a…Target microdroplet 202b…Waste microdroplets L…Center axis D213, D214…distance D211, D212, D2122…Inner diameter

Claims

1. A dyeing process includes dyeing a liquid specimen containing a plurality of bioparticles with a fluorescent dyeing process, thereby giving a fluorescent color to at least one of the plurality of bioparticles, thereby making the bioparticle a target bioparticle; an analysis step of storing the fluorescently stained liquid specimen in a microdroplet generating device and obtaining an image of the liquid specimen stored in the microdroplet generating device in real time using a camera device; A step of dropping a target microdroplet containing at least one of the target bioparticles onto a biochip from the microdroplet generating device according to the real-time image, and capturing the at least one of the target bioparticles contained in the target microdroplet on the biochip; a washing step for removing the fluorescent color of at least one of the target bioparticles captured on the biochip in a washing process; a characterization expressing step, in which at least one of the target bioparticles captured on the biochip is fluorescently stained N times by the fluorescent staining process and the washing process, and a plurality of fluorescent images corresponding to N types of biological characterization expressions are obtained by a recording device, where N is a positive integer between 2 and 50; The microdroplet generating device comprises a hollow needle, a container for containing the liquid sample, and an upper piezoelectric member disposed in the container; The hollow needles are connected to a bottom surface of the container so as to be in communication with each other; A method for analyzing biological particles, characterized in that the upper piezoelectric member vibrates the container, thereby arranging a plurality of the biological particles in the container along a predetermined path passing through the hollow needle.

2. The micro-droplet generator further includes a bottom piezoelectric member disposed on an outer surface of the hollow needle; The bioparticle analysis method of claim 1, wherein in the capture step, the microdroplet generating device squeezes the hollow needle with the bottom piezoelectric member, causing the liquid sample to flow out and pass through the free end of the hollow needle to form the microdroplets.

3. and during said analyzing step, said camera device includes obtaining a real-time image of the liquid specimen at said free end; The biological particle analysis method of claim 2, wherein in the capture process, a control device electrically connected to the bottom piezoelectric member and the camera device drives the bottom piezoelectric member when at least one of the target biological particles is at the free end by the instantaneous image.

4. In the staining step, the fluorescently stained liquid sample is contained in a sample container placed on a stand, In the analyzing step, the microdroplet generating device sucks the liquid sample from the sample container at the free end by moving relative to the platform; the biochip and the waste container are placed on the platform; The bioparticle analysis method according to claim 2 , wherein the microdroplet generating device drops the target microdroplets onto the biochip and drops waste microdroplets not containing the target bioparticles into the waste container by moving relative to the mounting table.

5. The biochip comprises a bottom layer, a plurality of capture arms connected to the bottom layer and spaced apart from each other, and a surface modification layer formed on the ends of the plurality of capture arms; The bioparticle analysis method according to claim 1 , wherein in the capturing step, the biochip captures at least one of the target bioparticles with the plurality of capture arms and the surface modification layer.

6. The method for analyzing bioparticles according to claim 5, wherein the surface modification layer is further limited to an arginylglycylaspartic acid (RGD) peptide layer.

7. The bioparticle analysis method according to claim 5 , wherein in the capturing step, the biochip holds at least one of the target bioparticles by elastically swinging at least two of the capture arms relative to the bottom layer.

Images