Cell activity detection system and method

Abstract

The invention discloses a cell viability detection system and method.The cell viability detection system comprises a micro-fluidic chip module, an optical detection module, an electrochemical detection module, a data processing and analyzing module and a control module, and the micro-fluidic chip module is used for monitoring physical and chemical signal changes related to cell viability in real time; compared with the prior art, the method has the following advantages that optical and electrochemical detection technologies are combined, cell activity information is obtained from multiple dimensions, the accuracy and reliability of cell activity detection are remarkably improved through comprehensive analysis, the real cell activity state can be reflected more accurately, and the accuracy and reliability of cell activity detection are improved. The micro-fluidic chip technology realizes rapid mixing and reaction of cells and reagents, and meanwhile, the automatic detection and data processing flow greatly shortens the detection time, improves the detection efficiency and meets the high-throughput detection requirement.

Description

Technical Field

[0001] The present invention relates to the technical field of cell detection, and particularly relates to a cell viability detection system and method. Background Art

[0002] Cell viability detection plays a crucial role in fields such as biomedical research, drug development, and cell therapy. Accurately evaluating cell viability is essential for understanding the physiological state of cells, screening the efficacy of drugs, and ensuring the safety and effectiveness of cell therapy.

[0003] Traditional cell viability detection methods have many drawbacks. For example, although the trypan blue staining method is simple to operate, it can only distinguish dead cells from live cells, cannot accurately reflect the degree of cell viability, and is highly subjective, with results easily affected by human factors. The MTT colorimetric method indirectly reflects cell viability by detecting the activity of succinate dehydrogenase in cell mitochondria. However, this method is easily affected by factors such as the cell metabolic state and drug interference, resulting in inaccurate detection results; at the same time, the detection process is cumbersome, and organic solvents are required to dissolve the formazan product, which not only pollutes the environment but also increases the experimental cost and time. In addition, although flow cytometry can quickly analyze a large number of cells, the instrument is expensive, sample preparation is complex, professional operators are required, and the detection throughput is limited, making it difficult to meet the high-throughput detection requirements. With the in-depth study of cells and the development of related industries, there is an urgent need for a more accurate, efficient, convenient, and high-throughput cell viability detection system and method. Therefore, a cell viability detection system and method are provided to solve the above problems. Summary of the Invention

[0004] To solve the problems in the above background art, the technical solution adopted by the present invention to solve the technical problems is: a cell viability detection system, which includes: a microfluidic chip module, an optical detection module, an electrochemical detection module, a data processing and analysis module, and a control module. The microfluidic chip module is used to monitor the changes in physical and chemical signals related to cell viability in real time. The optical detection module is used to detect the fluorescence signals emitted by cells after being stained with viability detection reagents, and different fluorescence markers are used to distinguish different viability states of cells. The electrochemical detection module is used to detect the electrochemical signals generated during the cell metabolism process in real time, directly reflecting the viability state of cells. The data processing module is used to receive the data from the optical detection module and the electrochemical detection module, and use advanced algorithms to process and analyze the image data and electrochemical signals.

[0005] As a preferred technical solution of the present invention, the microfluidic chip module includes a cell injection channel, a reagent injection channel, a reaction microchamber, and a detection microchannel. The cell injection channel and the reagent injection channel are controlled by microvalves to ensure that cells and reagents are mixed in precise proportions.

[0006] As a preferred technical solution of the present invention, the reaction microchamber is a place for cells to react with active reagents, and the surface of the detection microchannel is modified with a sensing material for real-time monitoring of physical and chemical signal changes related to cell activity.

[0007] As a preferred technical solution of the present invention, the optical detection module includes a fluorescence microscope and a phase contrast microscope. The fluorescence microscope is used to detect the fluorescence signals emitted by cells after being stained with active detection reagents, and different fluorescence markers are used to distinguish different activity states of cells. The phase contrast microscope is used to observe the morphological changes of cells, and combined with the fluorescence signals, the cell activity can be evaluated more comprehensively.

[0008] As a preferred technical solution of the present invention, the fluorescence microscope is of high sensitivity type, the phase contrast microscope is of high resolution type, and the optical detection module includes an autofocus image acquisition module, which can quickly and accurately acquire cell images.

[0009] As a preferred technical solution of the present invention, the electrochemical detection module uses a microelectrode array, and a plurality of microelectrodes are integrated at the bottom of the detection microchannel of the microfluidic chip. The electrochemical signals generated during the cell metabolism process can be detected in real time through the microelectrodes, so as to directly reflect the activity state of cells.

[0010] As a preferred technical solution of the present invention, the data processing and analysis module establishes a cell activity evaluation model, comprehensively integrates multi-dimensional data such as fluorescence intensity, cell morphological parameters, and electrochemical signals, accurately calculates the cell activity index, and generates a detailed detection report.

[0011] As a preferred technical solution of the present invention, the data processing and analysis module has data storage and comparative analysis functions, which is convenient for users to compare and study the detection results of different batches.

[0012] As a preferred technical solution of the present invention, the control module is used to control the opening and closing of microvalves in the microfluidic chip module, the flow rate control of reagents and cells, and the parameter setting of the optical detection module and the electrochemical detection module.

[0013] A detection method for a cell activity detection system includes the following steps:

[0014] Step 1: First, collect the cell samples to be detected, suspend them in a suitable cell culture medium, and at the same time, prepare corresponding cell activity detection reagents according to the detection requirements, such as fluorescent dyes and metabolic substrates;

[0015] Step 2: The cell sample and the detection reagent are respectively injected into the cell injection channel and the reagent injection channel of the microfluidic chip module through the control module. The micro-valve is opened to allow the cells and the reagent to flow into the reaction microchamber at a preset ratio and be fully mixed under the action of the micro-stirring structure;

[0016] Step 3: The cells and the reagent react in the reaction microchamber. As time goes by, the physical and chemical signals related to cell activity change. The optical detection module real-time collects the fluorescence images and phase contrast images of the cells, and the electrochemical detection module synchronously monitors the electrochemical signals generated by cell metabolism;

[0017] Step 4: The data collected by the optical detection module and the electrochemical detection module are transmitted to the data processing and analysis module. The data is preprocessed to remove noise and interference signals. Then, image recognition algorithms are used to analyze the cell images to extract cell morphological features and fluorescence intensity information;

[0018] Step 5: Electrochemical signal analysis algorithms are used to analyze the electrochemical data. The multi-dimensional data is input into the cell activity evaluation model to calculate the cell activity index and generate a test report.

[0019] The present invention has the following advantages: The present invention combines optical and electrochemical detection technologies to obtain cell activity information from multiple dimensions. Through comprehensive analysis, the accuracy and reliability of cell activity detection are significantly improved, and it can more accurately reflect the true activity state of cells;

[0020] The microfluidic chip technology realizes the rapid mixing and reaction of cells and reagents. At the same time, the automated detection and data processing processes greatly shorten the detection time, improve the detection efficiency, meet the high-throughput detection requirements. Users can complete complex detection processes with simple operations, reducing the requirements for the professional skills of operators. Compared with traditional detection technologies such as flow cytometry, the detection system of the present invention does not require expensive large-scale instrument equipment and uses less reagents, effectively reducing the detection cost.

[0021] It can not only be used for conventional cell activity detection, but also be applied to fields such as drug screening and cytotoxicity testing, and has broad application prospects. Description of the Drawings

[0022] Figure 1 is a schematic diagram of the cell activity detection system of the preferred embodiment of the present invention;

[0023] Figure 2 is a schematic diagram of the detection method of the cell activity detection system of the preferred embodiment of the present invention. Detailed Embodiments

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention. In the following description, many specific details are set forth to facilitate a thorough understanding of the present invention. However, the present invention may be implemented in other ways different from those described herein. Therefore, the protection scope of the present invention is not limited by the specific embodiments disclosed below.

[0025] In order to more clearly understand the above objects, features, and advantages of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0026] Please refer to Figure 1 - Figure 2 , a cell viability detection system of the present invention includes: a microfluidic chip module, an optical detection module, an electrochemical detection module, a data processing and analysis module, and a control module. The microfluidic chip module is used to monitor the changes in physical and chemical signals related to cell viability in real time. The optical detection module is used to detect the fluorescence signals emitted by cells after being stained with viability detection reagents, and different fluorescence markers are used to distinguish different viability states of cells. The electrochemical detection module is used to detect the electrochemical signals generated during the cell metabolism process in real time, directly reflecting the viability state of cells. The data processing module is used to receive the data from the optical detection module and the electrochemical detection module, and uses advanced algorithms to process and analyze the image data and electrochemical signals. Multilayer microfluidic chips are fabricated on materials such as polydimethylsiloxane (PDMS) using microfabrication processes such as photolithography and etching. The dimensions of the microchannels and reaction microcavities are precisely controlled to ensure the flow and reaction effects of cells and reagents in the chip. When modifying the sensing material on the chip surface, methods such as chemical vapor deposition are used to ensure that the sensing material is uniformly and firmly attached to the chip surface. The prepared multilayer microfluidic chips are assembled, and the cell injection channel, reagent injection channel, and detection microchannel are connected, and microvalves and microstirring structures are installed. The opening and closing of the microvalves and the flow rate are adjusted through the control system to ensure that cells and reagents can accurately and stably flow into the reaction microcavity.

[0027] Among them, the microfluidic chip module includes a cell injection channel, a reagent injection channel, a reaction microchamber, and a detection microchannel. The cell injection channel and the reagent injection channel are controlled by microvalves to ensure the precise mixing of cells and reagents. The reaction microchamber is used as the reaction site for cells and active reagents. The surface of the detection microchannel is modified with a sensing material to monitor the changes in physical and chemical signals related to cell activity in real time. The optical detection module includes a fluorescence microscope and a phase-contrast microscope. The fluorescence microscope is used to detect the fluorescence signals emitted by cells after being stained with active detection reagents, and different fluorescence markers are used to distinguish different activity states of cells. The phase-contrast microscope is used to observe the morphological changes of cells. Combining with the fluorescence signals, the cell activity can be evaluated more comprehensively. The fluorescence microscope is of high sensitivity type, and the phase-contrast microscope is of high resolution type. The optical detection module includes an autofocus image acquisition module, which can quickly and accurately obtain cell images. The high-sensitivity fluorescence microscope and the high-resolution phase-contrast microscope are installed on a stable optical platform for optical path calibration and lens debugging to ensure clear and stable imaging quality. Set the excitation light wavelength and fluorescence filter combination of the fluorescence microscope to match the fluorescence characteristics of the cell activity detection reagent. Image acquisition system settings: Install an autofocus and image acquisition system, and set image acquisition parameters such as exposure time and acquisition frequency. Through software programming, automatic acquisition and storage of images are realized to ensure that cell images can be obtained in real time and accurately during the detection process.

[0028] The electrochemical detection module uses a microelectrode array. Multiple microelectrodes are integrated at the bottom of the detection microchannel of the microfluidic chip. Through the microelectrodes, the electrochemical signals generated during the cell metabolism process can be detected in real time, thereby directly reflecting the activity state of cells. The data processing and analysis module establishes a cell activity evaluation model, comprehensively integrates multi-dimensional data such as fluorescence intensity, cell morphological parameters, and electrochemical signals, accurately calculates the cell activity index, and generates a detailed detection report. The data processing and analysis module has data storage and comparative analysis functions, which is convenient for users to compare and study the detection results of different batches. The control module is used to control the opening and closing of the microvalves in the microfluidic chip module, the flow rate control of reagents and cells, and the parameter settings of the optical detection module and the electrochemical detection module. The control module adopts a human-computer interaction interface, and users can easily input detection parameters and instructions through the interface to realize the automated operation of the detection process. Connect the microelectrode array to an electrochemical detection instrument for system calibration and parameter settings. Through standard solution testing, verify the performance of the microelectrode array to ensure that the electrochemical signals generated by cell metabolism can be accurately detected.

[0029] A detection method for a cell activity detection system includes the following steps:

[0030] Step 1: First, collect the cell sample to be tested and suspend it in a suitable cell culture medium. At the same time, prepare the corresponding cell activity detection reagents, such as fluorescent dyes and metabolic substrates, according to the detection requirements;

[0031] Step 2: inject the cell sample and the detection reagent into the cell injection channel and the reagent injection channel of the microfluidic chip module respectively through the control module, open the microvalve, and allow the cells and the reagent to flow into the reaction microchamber according to the preset ratio, and be fully mixed under the action of the micro-stirring structure;

[0032] Step 3: The cells and reagents react in the reaction microcavity. Over time, the physical and chemical signals related to cell activity change. The optical detection module collects fluorescence images and phase contrast images of the cells in real time, and the electrochemical detection module synchronously monitors the electrochemical signals generated by cell metabolism.

[0033] Step 4: The data collected by the optical detection module and the electrochemical detection module are transmitted to the data processing and analysis module, and the data are preprocessed to remove noise and interference signals. Then, the cell image is analyzed using an image recognition algorithm to extract cell morphological characteristics and fluorescence intensity information;

[0034] Step 5: Use electrochemical signal analysis algorithms to analyze electrochemical data, input multi-dimensional data into the cell activity evaluation model, calculate the cell activity index, and generate a test report.

[0035] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the attached claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0036] Other parts of the present invention not described in detail belong to the prior art and will not be described in detail here.

[0037] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A cell viability detection system, characterized in that, include: Microfluidic chip module, optical detection module, electrochemical detection module, data processing and analysis module, control module, the microfluidic chip module is used to monitor the changes in physical and chemical signals related to cell activity in real time, the optical detection module is used to detect the fluorescent signals emitted by the cells after being stained with activity detection reagents, and to distinguish the different activity states of the cells by different fluorescent markers, the electrochemical detection module is used to detect the electrochemical signals generated during the cell metabolism process in real time, and directly reflect the activity state of the cells, and the data processing module is used to receive data from the optical detection module and the electrochemical detection module, and to process and analyze the image data and electrochemical signals using advanced algorithms.

2. The cell activity detection system according to claim 1, wherein The microfluidic chip module includes a cell injection channel, a reagent injection channel, a reaction microchamber and a detection microchannel. The cell injection channel and the reagent injection channel are controlled by microvalves to ensure that cells and reagents are mixed in a precise proportion.

3. A cell viability detection system according to claim 2, characterized in that, The reaction microcavity is used as a place for cells to react with active reagents, and the surface of the detection microchannel is modified with sensing materials to monitor the physical and chemical signal changes related to cell activity in real time.

4. The cell activity detection system according to claim 1, wherein The optical detection module includes a fluorescence microscope and a phase contrast microscope. The fluorescence microscope is used to detect the fluorescence signal emitted by the cells after being stained with an activity detection reagent, and to distinguish the different activity states of the cells through different fluorescent markers. The phase contrast microscope is used to observe the morphological changes of the cells and, combined with the fluorescence signal, to more comprehensively evaluate the cell activity.

5. The cell activity detection system according to claim 4, wherein The fluorescence microscope is of high sensitivity, the phase contrast microscope is of high resolution, and the optical detection module includes an autofocus image acquisition module, which can quickly and accurately acquire cell images.

6. The cell activity detection system according to claim 1, wherein The electrochemical detection module utilizes a microelectrode array to integrate multiple microelectrodes at the bottom of the detection microchannel of the microfluidic chip. The microelectrodes can detect the electrochemical signals generated during cell metabolism in real time, thereby directly reflecting the activity state of the cells.

7. The cell activity detection system according to claim 1, wherein The data processing and analysis module establishes a cell activity evaluation model, integrates fluorescence intensity, cell morphology parameters, and electrochemical signal multi-dimensional data, accurately calculates the cell activity index, and generates a detailed detection report.

8. The cell viability detection system according to claim 1, wherein The data processing and analysis module has data storage and comparative analysis functions, which is convenient for users to compare and study the test results of different batches.

9. The cell activity detection system according to claim 1, characterized in that, The control module is used to control the opening and closing of the microvalve in the microfluidic chip module, the flow rate control of reagents and cells, and the parameter setting of the optical detection module and the electrochemical detection module.

10. The detection method of a cell activity detection system according to any one of claims 1-9, characterized in that, The following steps are involved: Step 1: First, collect the cell sample to be tested and suspend it in a suitable cell culture medium. At the same time, prepare the corresponding cell activity detection reagents, such as fluorescent dyes and metabolic substrates, according to the detection requirements; Step 2: inject the cell sample and the detection reagent into the cell injection channel and the reagent injection channel of the microfluidic chip module respectively through the control module, open the microvalve, and allow the cells and the reagent to flow into the reaction microchamber according to the preset ratio, and be fully mixed under the action of the micro-stirring structure; Step 3: The cells react with the reagents in the reaction microchamber. As time goes by, the physical and chemical signals related to cell viability change. The optical detection module collects the fluorescence images and phase-contrast images of the cells in real time, and the electrochemical detection module synchronously monitors the electrochemical signals generated by cell metabolism; Step 4: Transmit the data collected by the optical detection module and the electrochemical detection module to the data processing and analysis module, preprocess the data to remove noise and interference signals, and then use image recognition algorithms to analyze the cell images and extract cell morphological features and fluorescence intensity information; Step 5: Use electrochemical signal analysis algorithms to analyze the electrochemical data, input the multi-dimensional data into the cell viability assessment model, calculate the cell viability index, and generate a detection report.

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