Cell flow cell, cell detection device and method

Abstract

The present invention provides a cell flow cell, a cell detection device and a method, wherein the cell flow cell comprises: a body having an inlet and an outlet, a plurality of flow channels suitable for cells to pass through and grooves suitable for accommodating cells distributed in an array, the flow channels are used to connect adjacent grooves; the inlet and the outlet are respectively connected to the grooves, the depth of the grooves is greater than the depth of the flow channels; a plurality of openings are distributed in an array at the upper end of the body and are located on the upper side of the grooves; the grooves are connected to the outside world through the openings. The present invention has the advantages of accurate detection and automation.

Description

Technical Field

[0001] The present invention relates to cell analysis, and in particular to a cell flow cell, a cell detection device and a method. Background Art

[0002] Single-cell technology has developed rapidly in recent years. With the continuous development of technologies such as single-cell sequencing and single-cell transcriptomics, humans have gained a deep understanding of the heterogeneity of cell populations at the DNA and RNA levels. At the protein level, flow cytometry has always been the most commonly used single-cell analysis method, but due to problems such as cross-linking, flow cytometry can usually only detect 6-10 proteins at the same time.

[0003] Mass cytometry is a flow technology that uses the principle of mass spectrometry to perform multi-parameter detection on single cells. It inherits the high-speed analysis characteristics of traditional flow cytometers and has the high resolution capability of mass spectrometry detection. This fusion technology can simultaneously analyze more than 40 cell parameters at the single-cell level, greatly increasing the ability of flow cytometry analysis to evaluate complex cell systems and processes, and is a new development direction for flow cytometry. It uses metal element markers (antibodies) to mark or identify signal molecules on the cell surface and inside, then uses flow cytometry to separate single cells, and then uses ICP-MS to observe the atomic mass spectrum of single cells. Finally, the atomic mass spectrum data is converted into signal molecule data on the cell surface and inside, and the obtained data is analyzed by dimensionality reduction through computer analysis software, thereby achieving detailed observation of cell phenotypes and signal networks.

[0004] However, in mass spectrometry flow cytometry, after the cell samples are separated into single cells by the flow chamber and liquid flow system, they must all enter the mass spectrometry analysis in sequence, which is not conducive to quickly obtaining the analysis results of the target cells and has low efficiency. In the flow mass spectrometry based on the laser ablation method, there is a phenomenon of excessive sample desorption, which produces too many ions per unit time, thus causing a saturation effect and hindering mass spectrometry detection. Summary of the invention

[0005] In order to solve the deficiencies in the above-mentioned prior art solutions, the present invention provides a cell flow pool for dispersing and aggregating cells.

[0006] The objective of the present invention is achieved through the following technical solutions:

[0007] A cell flow cell, comprising:

[0008] A body, wherein the body has an inlet and an outlet, and has a plurality of flow channels suitable for cells to pass through and grooves suitable for accommodating cells distributed in an array, wherein the flow channels are used to connect adjacent grooves; the inlet and the outlet are respectively connected to the grooves, and the depth of the grooves is greater than the depth of the flow channels;

[0009] A plurality of openings are distributed in an array at the upper end of the body and are located on the upper side of the groove; the groove is connected to the outside through the openings.

[0010] Another object of the present invention is to provide a cell detection device based on the above-mentioned cell flow cell with accurate detection and high efficiency. This object of the invention is achieved through the following technical solutions:

[0011] A cell detection device, the cell detection device comprising a mass spectrometer; the cell detection device also comprises:

[0012] A sampling unit, the cell flow cell mentioned above in the sampling unit;

[0013] A detection unit, the detection unit is used to obtain the position of the groove of abnormal cells on the sampling unit;

[0014] A plurality of electrodes, which are arranged outside the groove and connected to a power source;

[0015] A transmission tube, one end of which is connected to the inlet of the mass spectrometer, and the other end of which receives ions emitted from the groove;

[0016] A driving unit drives the transmission tube or the sampling unit so that the transmission tube receives the ionized ions in any groove.

[0017] Another object of the present invention is to provide a cell detection method based on the above cell detection device, and this object of the invention is achieved through the following technical solutions:

[0018] According to the cell detection method of the above detection device, the cell detection method comprises the following steps:

[0019] (A1) The cell suspension enters the sampling unit; the cells stay in each groove, and the liquid flows through the flow channel and the groove and is discharged from the outlet;

[0020] (A2) the detection unit obtains the position of the groove where the abnormal cell is located on the sampling unit;

[0021] (A3) the electrode outside the groove where the abnormal cell is located discharges electricity, the cell in the groove is ionized, and the ions pass upward through the opening into the transmission tube;

[0022] (A4) The transfer tube delivers the ions to the mass spectrometer.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1. Avoid saturation effect;

[0025] The cell flow pool is specially designed. The cell suspension enters the interior, the cells are dispersed in each groove, and the excess liquid flows through the flow channel and the groove, and finally discharged from the outlet, which effectively prevents the cells from being over-concentrated in the groove and avoids the occurrence of saturation effect.

[0026] The design of the groove parameters (diameter and depth) prevents too many cells from gathering in a single groove, so that the cells are dispersed in each groove;

[0027] 2. High detection efficiency;

[0028] The detection unit detects abnormal cells, and then detects cells in the groove where the abnormal cells are located, without having to detect all cells, which significantly improves the detection efficiency;

[0029] 3. High detection accuracy;

[0030] The combination of the needle discharge electrode and the ring electrode forms a Taylor cone, which makes the discharge area more concentrated, increases the ion transmission efficiency, and improves the detection accuracy.

[0031] The design of the parameters (distance and angle) between the transmission tube and the cell flow cell improves the receiving efficiency of the ions emitted from the opening, and correspondingly improves the detection accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The disclosure of the present invention will become easier to understand with reference to the accompanying drawings. It is easy for those skilled in the art to understand that these drawings are only used to illustrate the technical solution of the present invention and are not intended to limit the scope of protection of the present invention. In the drawings:

[0033] Figure 1 is a flow chart of a cell detection method according to an embodiment of the present invention. DETAILED DESCRIPTION

[0034] Figure 1 The following description describes the optional embodiments of the present invention to teach those skilled in the art how to implement and reproduce the present invention. In order to teach the technical solution of the present invention, some conventional aspects have been simplified or omitted. It should be understood by those skilled in the art that variations or substitutions derived from these embodiments will be within the scope of the present invention. It should be understood by those skilled in the art that the following features can be combined in various ways to form multiple variations of the present invention. Thus, the present invention is not limited to the following optional embodiments, but is only limited by the claims and their equivalents.

[0035] Embodiment 1:

[0036] A cell detection device, comprising:

[0037] a mass spectrometer having an inlet;

[0038] A sampling unit, wherein the sampling unit adopts a cell flow cell, and the cell flow cell comprises: a body, wherein the body has an inlet and an outlet, and has a plurality of flow channels suitable for cells to pass through and grooves suitable for accommodating cells distributed in an array, wherein the flow channels are used to connect adjacent grooves; the inlet and the outlet are respectively connected to the grooves, and the depth of the grooves is greater than the depth of the flow channels; a plurality of openings, wherein the plurality of openings are distributed in an array at the upper end of the body and are located on the upper side of the grooves; and the grooves are connected to the outside through the openings;

[0039] A detection unit, the detection unit is used to obtain the position of the groove of abnormal cells on the sampling unit;

[0040] A plurality of electrodes, such as electrodes with pointed tips, are disposed outside the groove and connected to a power source;

[0041] A transmission tube, one end of which is connected to the inlet of the mass spectrometer, and the other end of which receives ions ionized by the cells in the groove after being discharged by the electrode;

[0042] A driving unit drives the transmission tube or the sampling unit so that the transmission tube receives the ionized ions in any groove.

[0043] In order to prevent the cells accumulated in the groove from escaping from the opening, further, the diameter of the opening is smaller than the diameter of the groove.

[0044] In order to accumulate cells appropriately and prevent excessive accumulation, the groove further has a diameter R∈[10 μm, 100 μm] and a depth H∈[5 μm, 50 μm].

[0045] In order to make the cells accumulate in the groove rather than in the flow channel, further, the inner wall of the flow channel has a hydrophobic layer, and the inner wall of the groove is a non-smooth surface.

[0046] In order to accurately and efficiently obtain the position of the groove where the abnormal cells are located, the detection unit further includes a light source, a detector, an analysis module and a light reflecting layer; the light reflecting layer is arranged on the bottom wall or the lower side of the groove of the sampling unit, and the detection light emitted by the light source illuminates the cells in the groove; the detector receives the reflected light of the detection light on the light reflecting layer, and outputs an electrical signal to the analysis module; the analysis module processes the electrical signal, determines whether the cells in each groove on the sampling unit are abnormal, and obtains the position of the groove where the abnormal cells are located.

[0047] In order to form a Taylor cone, further, one end of the transmission tube adjacent to the opening is provided with a grounding electrode; preferably, the grounding electrode is annular, and the diameter of the area enclosed by the annular electrode is smaller than the diameter of the groove.

[0048] In order to improve the efficiency of the transmission tube in receiving ions, further, the bottom end of the transmission tube is located on the upper side of the opening, and the distance between the bottom end and the opening is L∈[2mm, 10mm]; the angle between the central axis of the transmission tube and the top plane of the main body is

[0049] Figure 1 A flow chart of the cell detection method according to an embodiment of the present invention is given, that is, the working method of the detection device according to this embodiment, such as Figure 1 As shown, the cell detection method comprises the following steps:

[0050] (A1) The cell suspension enters the sampling unit from the inlet; the cells stay in each groove, the liquid flows through the flow channel and the groove, and is discharged from the outlet;

[0051] (A2) the detection unit obtains the position of the groove where the abnormal cell is located on the sampling unit;

[0052] (A3) the electrode outside the groove where the abnormal cell is located discharges electricity, the cell in the groove is ionized, and the ions pass upward through the opening and enter the transmission tube;

[0053] (A4) The transfer tube delivers the ions to the mass spectrometer.

[0054] Embodiment 2:

[0055] An example of application of the cell detection device and method according to Example 1 of the present invention in red blood cell detection.

[0056] In this application example, the sampling unit (cell flow cell) includes a first layer, a second layer, a third layer and a fourth layer stacked from top to bottom, and adjacent layers are connected by glue; the first layer has openings distributed in an array, and the diameter of the cylindrical opening is smaller than the diameter of the groove; the second layer has an inlet and an outlet on the side, and an array-distributed groove on the upper end, and the inner wall of the groove is a non-smooth surface suitable for accommodating one or more cells, and the diameter of the groove R∈[10μm, 100μm], such as 10μm, 30μm, 50μm, 80μm, 100μm, and the depth H∈ [5μm, 50μm], such as 5μm, 12μm, 24μm, 35μm, 50μm, adjacent grooves are connected by flow channels, the depth of the flow channels is less than that of the grooves and is suitable for the passage of cells, and the inner wall of the flow channel has a hydrophobic layer; the third layer is a light reflection layer; the fourth layer has pointed electrodes distributed in an array (the electrodes do not contact the cells inside the grooves), with the tips facing upward; the above four layers are used to form the following structure: the openings, grooves, reflection layers and pointed electrodes are arranged from top to bottom, so that the upper side of each groove is an opening, and the lower side is a reflection layer and a pointed electrode;

[0057] The detection unit includes a light source, a detector and an analysis module; the detection light emitted by the light source illuminates the cells in the groove, and the first layer and the second layer are suitable for transmitting the detection light; the detector receives the reflected light of the detection light on the light reflection layer, and outputs an electrical signal to the analysis module; the analysis module uses the electrical signal to obtain an image of the cell flow pool, and uses image recognition technology to determine whether the cells in each groove on the sampling unit are abnormal, and obtains the position of the groove where the abnormal cell is located;

[0058] A transmission tube, one end of which is connected to the inlet of the mass spectrometer, and the vacuum degree of the mass spectrometer is used to make the transmission tube negative pressure, so that the other end receives the ions of the cells in the groove after being ionized by the electrode discharge; the bottom end of the transmission tube is located on the upper side of the opening, and the distance between the bottom end and the opening is L∈[2mm, 10mm], such as 2mm, 5mm, 7mm or 10mm; the angle between the central axis of the transmission tube and the top plane of the main body is Such as 30 degrees, 45 degrees, 60 degrees or 90 degrees;

[0059] A grounding electrode is arranged at one end of the transmission tube adjacent to the opening, the grounding electrode is annular, and the diameter of the area surrounded by the annular electrode is smaller than the diameter of the groove;

[0060] The driving unit adopts a three-dimensional moving platform to drive the transmission tube so that the transmission tube receives the ionized ions in any groove.

[0061] The cell detection method according to the embodiment of the present invention, that is, the working method of the detection device according to the present embodiment, comprises the following steps:

[0062] (A1) The cell suspension enters the sampling unit through the inlet; the cells and the liquid flow through the flow channel and the groove, one or more cells stay in each groove, and the liquid flows through the flow channel and the groove and is discharged from the outlet;

[0063] (A2) the detection light emitted by the light source illuminates the cells in the groove, and the first layer and the second layer are suitable for transmitting the detection light;

[0064] The detector receives the reflected light of the detection light on the light reflection layer, and outputs an electrical signal to the analysis module;

[0065] The analysis module uses the electrical signal to obtain an image of the cell flow pool, and uses image recognition technology to determine whether the cell morphology in each groove is abnormal, and obtains the position of the groove where the abnormal cell is located;

[0066] (A3) the tip electrode at the lower side of the groove where the abnormal cell is located discharges, forming a Taylor cone between the tip electrode and the ring electrode, the cells in the groove are ionized, and the ions pass upward through the opening into the transmission tube;

[0067] (A4) Under the effect of negative pressure in the mass spectrometer, the transmission tube is under negative pressure, thereby transporting ions to the mass spectrometer, thereby obtaining the composition and content of abnormal cells.

[0068] Embodiment 3:

[0069] The application example of the cell detection device and method according to Example 1 of the present invention is different from that of Example 2 in that:

[0070] 1. The opening of the first layer is in the shape of a frustum, and the diameter of the opening increases from top to bottom, wherein the minimum diameter is smaller than the diameter of the groove, and the maximum diameter is not larger than the diameter of the groove.

[0071] 2. The angle between the transmission tube and the upper end surface of the first layer is 90 degrees.

[0072] 3. Instead of providing a separate light reflection layer, a light reflection film formed on the bottom wall of the groove is used.

[0073] Embodiment 4:

[0074] The application example of the cell detection device and method according to Example 1 of the present invention is different from that of Example 2 in that:

[0075] 1. Eliminate the ring electrode.

[0076] 2. The angle between the transmission tube and the upper end surface of the first layer is 90 degrees.

Claims

1. A cell detection device, comprising a mass spectrometer; characterized in that: The cell detection device also includes: A sampling unit, wherein the sampling unit adopts a cell flow cell; the cell flow cell comprises a body and a plurality of openings, the body having an inlet and an outlet, and having a plurality of flow channels suitable for cells to pass through and grooves suitable for accommodating cells distributed in an array, the flow channels being used to connect adjacent grooves; the inlet and the outlet are respectively connected to the grooves, the depth of the grooves being greater than the depth of the flow channels; the plurality of openings are distributed in an array at the upper end of the body and are located on the upper side of the grooves; the grooves are connected to the outside through the openings; A detection unit, the detection unit is used to obtain the position of the groove of abnormal cells on the sampling unit; A plurality of electrodes, wherein the electrodes are pointed electrodes, are arranged at the lower side of the groove and are connected to a power source; A transmission tube, one end of which is connected to the inlet of the mass spectrometer, and the other end of which receives ions from the cells in the groove after being ionized by the electrode; one end of the transmission tube facing the opening is provided with a grounding electrode; A driving unit, wherein the driving unit drives the transmission tube or the sampling unit so that the transmission tube receives the ionized ions in any groove; The detection unit includes a light source, a detector, an analysis module and a light reflecting layer, wherein the light reflecting layer is arranged on the bottom wall or the lower side of the groove of the sampling unit; the detection light emitted by the light source illuminates the cells in the groove; the detector receives the reflected light of the detection light on the light reflecting layer, and outputs an electrical signal to the analysis module; the analysis module processes the electrical signal to determine whether the cells in each groove on the sampling unit are abnormal, and obtains the position of the groove where the abnormal cells are located.

2. The cell detection device according to claim 1, characterized in that: The grounding electrode is ring-shaped, and the diameter of the area enclosed by the ring-shaped electrode is smaller than the diameter of the groove.

3. The cell detection device according to claim 1, characterized in that: The bottom end of the transmission tube is located above the opening, and the distance between the bottom end of the transmission tube and the opening is The angle between the central axis of the transmission tube and the top plane of the main body .

4. The cell detection device according to claim 1, characterized in that: The diameter of the opening is smaller than the diameter of the groove.

5. The cell detection device according to claim 1, characterized in that: The diameter of the groove ,depth .

6. The cell detection device according to claim 1, characterized in that: The inner wall of the flow channel has a hydrophobic layer, and the inner wall of the groove is a non-smooth surface.

7. The cell detection method according to any one of claims 1 to 6, wherein the cell detection method comprises the following steps: (A1) The cell suspension enters the sampling unit; the cells stay in each groove, and the liquid flows through the flow channel and grooves and is discharged from the outlet; (A2) The detection unit obtains the position of the groove where the abnormal cell is located on the sampling unit; (A3) The electrode outside the groove where the abnormal cell is located discharges electricity, the cell in the groove is ionized, and the ions pass upward through the opening into the transmission tube; (A4) The transfer tube transports the ions to the mass spectrometer.

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