Defective cell detection apparatus and method

Abstract

The application provides a device and method for detecting defective cells, which comprises a cell detector, a cell screening unit for screening cells of different diameters, a first switching module for selectively sending the screened cells of different diameters to a first channel, a first identification unit for identifying whether a single cell in the first channel is defective, a second switching module for switching the channel according to the result of the first identification unit, a positioning module for determining the position of a defective cell in a second channel, and a transfer unit for transferring the defective cell or its extract at the determined position to the cell detector. The application has the advantages of accurate detection and automation.

Description

Technical Field

[0001] This invention relates to cell analysis, and more particularly to a device and method for detecting defective cells. Background Technology

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

[0003] Mass cytometry is a flow cytometry technique that uses mass spectrometry principles to perform multi-parameter detection on single cells. It inherits the high-speed analysis capabilities of traditional flow cytometers while possessing the high-resolution detection capabilities of mass spectrometry. This fusion technique can simultaneously analyze more than 40 cellular parameters at the single-cell level, greatly increasing the ability of flow cytometry to analyze and evaluate complex cellular systems and processes, representing a new direction in the development of flow cytometry. It uses metal element markers (antibodies) to label or recognize signaling molecules on the cell surface and inside the cell. Then, flow cytometry is used to separate single cells, followed by ICP-MS observation of the atomic mass spectra of individual cells. Finally, the atomic mass spectra data are converted into data on cell surface and internal signaling molecules, and computer analysis software is used to perform dimensionality reduction analysis on the obtained data, thereby enabling detailed observation of cell phenotypes and signaling networks.

[0004] In mass cytometry, after cell samples are separated into individual cells by a flow chamber and liquid flow system, all of them must be sequentially analyzed by mass spectrometer. This process is not conducive to rapidly obtaining analytical results for the target cells, resulting in low efficiency. Furthermore, since each analysis targets only a single cell, the signal for analyzing a specific component within the cell is often weak. Summary of the Invention

[0005] To address the shortcomings of the existing technical solutions, the present invention provides a detection device for detecting defective cells that is accurate and automated.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] A device for detecting defective cells, comprising a cell detector; the device for detecting defective cells further comprising:

[0008] A cell screening unit, which filters out cells of different diameters;

[0009] The first switching module is used to selectively send cells of different diameters to the first channel.

[0010] The first identification unit is used to identify whether a single cell in the first channel has defects;

[0011] The second switching module switches channels according to the result of the first identification unit; if the identification result is a defective cell, the first channel is connected to the second channel; if the identification result is a non-defective cell, the first channel is connected to the third channel.

[0012] The positioning module is used to determine the position of the defective cell within the second channel;

[0013] A transfer unit for transferring defective cells or their extracts at a defined location to the cell detector.

[0014] Another object of the present invention is to provide a method for detecting defective cells, which is achieved through the following technical solution:

[0015] A method for detecting defective cells, comprising the following steps:

[0016] (A1) The cell suspension enters the cell screening unit, screening out cells with different diameters R. i Cells, i = 1, 2…n, n ≥ 2, and n ∈ N;

[0017] (A2) Select diameter R by switching. m Cells enter the first channel, 1≤m≤n, and m∈N;

[0018] (A3) Identify whether the cells in the first channel are defective;

[0019] If the cell is defective, the control switching module is activated to connect the first channel to the second channel, and the process proceeds to step (A4).

[0020] If the cell is defect-free, the control switching module is activated to connect the first channel to the third channel;

[0021] (A4) Use the positioning module to obtain the location of the defective cells in the second channel;

[0022] (A5) The extraction needle moves to the upper side of the location and then moves down; the extraction needle aspirates a single defective cell at the location;

[0023] (A6) Rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0025] 1. Accurate detection;

[0026] Cells of different diameters are screened out by the cell screening unit, which improves the accuracy of cell detection.

[0027] The recognition unit accurately identifies defective cells, improving the accuracy of defective cell detection.

[0028] During the detection process, multiple extraction needles pick up multiple defective cells (one extraction needle picks up only a single defective cell), and multiple defective cells are detected simultaneously using techniques such as mass spectrometry. If they are ionized at the same time, the detection signal intensity is increased, and the detection accuracy is improved accordingly.

[0029] 2. Automation;

[0030] Cell screening, identification, aspiration, transfer, and detection can all be automated, improving detection efficiency. Attached Figure Description

[0031] The disclosure of this invention will become more readily understood with reference to the accompanying drawings. It will be readily understood by those skilled in the art that these drawings are merely illustrative of the technical solutions of this invention and are not intended to limit the scope of protection of this invention. In the drawings:

[0032] Figure 1 This is a flowchart of a method for detecting defective cells according to an embodiment of the present invention. Detailed Implementation

[0033] Figure 1 The following description illustrates optional embodiments of the invention to teach those skilled in the art how to implement and reproduce the invention. Some conventional aspects have been simplified or omitted for the purpose of teaching the invention. Those skilled in the art should understand that variations or substitutions derived from these embodiments will be within the scope of the invention. Those skilled in the art should understand that the following features can be combined in various ways to form multiple variations of the invention. Therefore, the invention is not limited to the following optional embodiments, but is defined only by the claims and their equivalents.

[0034] Example 1:

[0035] A device for detecting defective cells, the device comprising:

[0036] Cell detection instruments, such as mass spectrometers;

[0037] A cell screening unit, which filters out cells of different diameters;

[0038] The first switching module is used to selectively send cells of different diameters to the first channel.

[0039] The first identification unit is used to identify whether a single cell in the first channel has defects;

[0040] The second switching module switches channels according to the result of the first identification unit; if the identification result is a defective cell, the first channel is connected to the second channel so that the defective cell enters the second channel; if the identification result is a non-defective cell, the first channel is connected to the third channel.

[0041] The positioning module is used to determine the position of the defective cell within the second channel;

[0042] A transfer unit for transferring defective cells or their extracts at a defined location to the cell detector.

[0043] To accurately screen cells of different diameters, the cell screening unit further includes:

[0044] At least two interception modules are stacked one on top of the other, and each interception module includes a groove; from top to bottom, the width of the groove of the interception module decreases and they are connected; the input end of the first switching module is connected to the groove outlet of each interception module.

[0045] To improve the screening effect of cells with different diameters and to drive cell forward movement, the cell screening unit further includes:

[0046] A vibrator connected to the at least two interception modules.

[0047] To accurately identify defective cells, the first identification unit further includes:

[0048] A light source, wherein the light source is disposed on the side of the first channel;

[0049] A detector that receives light emitted by the light source and passing through the first channel to obtain an image of the cell;

[0050] The identification module identifies whether cells have defects based on the image.

[0051] To accurately locate the position of defective cells within the second channel, the positioning module is further positioned below the second channel to obtain an image of the defective cell and identify its location; or,

[0052] The positioning module employs a groove suitable for accommodating a single defective cell.

[0053] To transfer defective cells within the second channel, the transfer unit further includes:

[0054] One or more extraction needles, said extraction needles being used to aspirate single cells or extracts thereof;

[0055] A drive module, which is used to drive the extraction needle to move up and down;

[0056] A rotating module is used to rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector.

[0057] To further confirm that the cells in the second channel are defective, a light source is further positioned between the extraction needles. The positioning module receives the light emitted by the light source that passes through the first channel, obtains an image of the cell, and identifies whether the cell is defective.

[0058] To temporarily store defective cells, the defective cell detection device further includes:

[0059] A valve is provided on the second channel between the second switching module and the positioning module.

[0060] Figure 1 A flowchart of the defective cell detection method according to an embodiment of the present invention is provided, that is, the working method of the detection device according to this embodiment, such as... Figure 1 As shown, the method for detecting defective cells includes the following steps:

[0061] (A1) The cell suspension enters the cell screening unit, screening out cells with different diameters R. i Cells, i = 1, 2…n, n ≥ 2, and n ∈ N;

[0062] (A2) Select diameter R by switching. m Cells enter the first channel, 1≤m≤n, and m∈N;

[0063] (A3) Use the first recognition unit to identify whether the cells in the first channel have defects, such as through cell imaging, image analysis (machine vision), etc.

[0064] If the cell is defective, the control switching module is activated to connect the first channel to the second channel, and the process proceeds to step (A4).

[0065] If the cell is defect-free, the control switching module is activated to connect the first channel to the third channel;

[0066] (A4) Use the positioning module to obtain the location of the defective cells in the second channel;

[0067] (A5) The extraction needle moves to the upper side of the said position and then moves down; the extraction needle aspirates a single defective cell at the said position: such as a single extraction needle aspirating a single defective cell, or multiple extraction needles aspirating multiple defective cells;

[0068] (A6) Rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector, thereby analyzing the substances in the defective cells.

[0069] To facilitate accurate aspiration of defective cells by the extraction needle, the defective cells are further positioned as follows:

[0070] When the valve on the second channel is closed, defective cells entering the second channel are blocked by the valve, and the defective cells accumulate and are temporarily stored.

[0071] After the single defective cell in the aspiration position is taken away by the extraction needle, the valve is opened, and one or more temporarily stored defective cells pass through the valve and enter the aspiration position;

[0072] Repeat the above process so that the temporarily stored defective cells enter the aspiration position in sequence;

[0073] Alternatively, defective cells entering the second channel are blocked and temporarily stored in sequence; one or more defective cells at the front of the line are drawn up by the extraction needle, and the other temporarily stored defective cells move forward.

[0074] Example 2:

[0075] Application examples of the defective cell detection device and method according to Embodiment 1 of the present invention.

[0076] In this application example, the cell screening unit uses a three-layer stacked interception module. The interception module has straight grooves. From top to bottom, the width of each groove decreases and they are connected vertically, so that the diameter of the intercepted cells becomes smaller and smaller. An ultrasonic vibrator is connected to the interception module. The first switching module uses a multi-way valve, so that the first channel selectively connects to any straight groove, that is, it receives cells of various diameters.

[0077] The first identification unit uses a combination of a light source, a detector, and an identification module. The light source and the detector are located on both sides of the first channel. The identification module uses machine vision to identify defective cells. If the image of a cell is compared with the image of a normal cell, and the deviation is large, it is identified as a defective cell.

[0078] The second switching module uses a three-way valve to selectively connect the second channel and the third channel. The selection principle is: if the result of the first identification unit is that the cell is defective, then the first channel and the second channel are connected by switching; otherwise, the first channel and the third channel are connected.

[0079] The positioning module uses a groove to accommodate a single defective cell and is located within the second channel;

[0080] Multiple extraction needles are mounted on a first rotating module; the first rotating module is mounted on a second rotating module, with the axis of rotation of the second rotating module being horizontal; the axis of rotation of the first rotating module is vertically aligned with the axis of rotation of the second rotating module.

[0081] The cell detector uses a mass spectrometer and has an ion inlet;

[0082] The first, second, and third channels mentioned above are all transparent tubes. The upper end of the second channel has an opening to facilitate the extraction needle to enter the second channel and aspirate defective cells.

[0083] The method for detecting defective cells in this embodiment includes the following steps:

[0084] (A1) The cell suspension enters the cell screening unit. The ultrasonic vibrator drives the retention module to vibrate. The suspension flows from top to bottom, screening out three different diameter cells. The largest diameter cell is in the groove of the uppermost retention module, and the smallest diameter cell is in the groove of the lowermost retention module.

[0085] (A2) Through the automatic switching of the multi-way valve, the cell with the largest diameter (that is, the cell in the groove of the uppermost interception module) is selected to enter the first channel;

[0086] (A3) The first recognition unit is used to identify whether the cells in the first channel have defects, and the defective cells are automatically identified through cell imaging and image analysis technology;

[0087] If the cell has defects, then in time The three-way valve is then automatically controlled to connect the first channel to the second channel. The defective cell enters the second channel through the three-way valve, and the three-way valve is switched after time t0+t1, closing the connection between the first and second channels. L is the distance between the first identification unit and the three-way valve in the first channel, v is the speed of cell movement, and t1 is the time it takes for the defective cell to pass through the three-way valve. That is, the three-way valve maintains the connection between the first and third channels during the normal time. Only after the defective cell is identified is the three-way valve used to connect the first and second channels; and then proceed to step (A4).

[0088] If the cells are defect-free, the three-way valve is switched so that the first channel connects to the third channel, allowing defect-free cells to enter the third channel.

[0089] (A4) The location of the defective cells in the second channel is obtained using the groove;

[0090] (A5) The extraction needle moves to the upper side of the position (at this time the axis of the first rotating module is vertical) and moves down. The extraction needle picks up a single defective cell at the position and then moves up. Defective cells continuously pass through the three-way valve and enter the groove of the second channel. The first rotating module rotates so that different extraction needles pick up the defective cells in the groove.

[0091] (A6) After all the cells with the largest diameter have passed through the first recognition unit, the second rotating module rotates around the horizontal axis so that the outlet of the extraction needle corresponds to the inlet of the cell detector.

[0092] The electrodes on each extraction needle discharge, and multiple defective cells are simultaneously ionized and enter the inlet of the mass spectrometer for analysis, thereby analyzing the substances inside the defective cells.

[0093] Example 3:

[0094] An application example of the defective cell detection device and method according to Embodiment 1 of the present invention differs from Embodiment 2 in that:

[0095] A valve is installed in the second channel, which is located between the three-way valve and the positioning groove. Defective cells passing through the three-way valve are temporarily stored in the second channel between the three-way valve and the valve. The cells are arranged in sequence, and the position of the cell at the front adjacent to the valve is determined, so that the valve indirectly has the function of a positioning module. As the first rotating module rotates, different extraction needles continuously pick up the defective cells at the front (when the foremost defective cell is picked up, the cells behind it move forward).

[0096] Example 4:

[0097] An application example of the defective cell detection device and method according to Embodiment 1 of the present invention differs from Embodiment 2 in that:

[0098] A light source is placed between the extraction needles, and a detector is placed on the lower side of the second channel to obtain an image of the defective cells in the second channel. The image is processed by the positioning module to obtain the position of the defective cells in the second channel. The extraction needles are then used to extract the defective cells at that position.

[0099] Example 5:

[0100] An application example of the defective cell detection device and method according to Embodiment 1 of the present invention differs from Embodiment 2 in that:

[0101] 1. A valve is installed on the second channel, which is located between the three-way valve and the positioning groove. Defective cells passing through the three-way valve are temporarily stored in the second channel between the three-way valve and the valve. By opening and closing the valve, the temporarily stored defective cells pass through the valve one by one (i.e., only one cell passes through the valve at a time) and enter the positioning groove one by one. As the first rotating module rotates, different extraction needles pick up the individual defective cells in the groove at different times.

[0102] 2. Set up a second identification unit with the same structure as the first identification unit, wherein the light source is set between the extraction needles and the detector is set on the lower side of the second channel; using the second identification unit, confirm again whether the defective cells in the positioning groove in the second channel are defective. If the defect is verified again, control the extraction needle to draw them in. If the defect is verified, discharge them from the second channel and no longer detect them.

Claims

1. A device for detecting defective cells, the device comprising a cell detector; characterized in that, The defective cell detection device also includes: A cell screening unit, wherein the cell screening unit filters out cells of different diameters; The first switching module is used to selectively send cells of different diameters to the first channel. The first identification unit is used to identify whether a single cell in the first channel has defects; The second switching module uses a three-way valve. The second switching module switches channels according to the result of the first identification unit. If the identification result is a defective cell, the first channel is connected to the second channel. If the identification result is a non-defective cell, the first channel is connected to the third channel. The second channel is a transparent pipe, and part of its upper end has an opening to facilitate the transfer unit to enter the second channel to absorb defective cells. A positioning module is used to determine the position of defective cells in the second channel; a valve is set on the second channel so that the defective cells passing through the second switching module are arranged sequentially between the valve and the second switching module, and the position of the cell at the front adjacent to the valve is determined, and the valve indirectly has the function of the positioning module; A transfer unit is used to transfer defective cells located at the foremost point of the valve in the second channel to the cell detector; the transfer unit includes: Multiple extraction needles are mounted on a first rotating module; the first rotating module is mounted on a second rotating module, the second rotating module having a horizontal axis of rotation; the axis of rotation of the first rotating module is perpendicular to the axis of rotation of the second rotating module. When the first rotating module rotates, different extraction needles continuously pick up the foremost defective cells. After the foremost defective cell is picked up, the cells behind it move forward. When the second rotating module rotates around the axis of rotation, the outlet of the extraction needle corresponds to the ion inlet of the cell detector, and multiple defective cells picked up by multiple extraction needles are simultaneously ionized. The cell detector is a mass spectrometer.

2. The defective cell detection device according to claim 1, characterized in that, The cell screening unit includes: At least two interception modules are stacked one on top of the other, and each interception module includes a groove; from top to bottom, the width of the groove of the interception module decreases and they are connected; the input end of the first switching module is connected to the groove outlet of each interception module.

3. The defective cell detection device according to claim 2, characterized in that, The cell screening unit further includes: A vibrator connected to the at least two interception modules.

4. The defective cell detection device according to claim 1, characterized in that, The first identification unit includes: A light source, wherein the light source is disposed on the side of the first channel; A detector that receives light emitted by the light source and passing through the first channel to obtain an image of the cell; The identification module identifies whether cells have defects based on the image.

5. The method for detecting defective cells implemented by the detection device according to claim 1, wherein the method for detecting defective cells includes the following steps: (A1) The cell suspension enters the cell screening unit to screen out cells with different diameters R. i Cells, i = 1, 2…n, n ≥ 2, and n ∈ N; (A2) By switching through the first switching unit, the diameter R is selected. m Cells enter the first channel, 1≤m≤n, and m∈N; (A3) Identify whether the cells in the first channel are defective; If the cell is defective, the second switching module is controlled to connect the first channel to the second channel, and the process proceeds to step (A4). If the cell is defect-free, the second switching module is controlled to connect the first channel to the third channel; (A4) The defective cells entering the second channel are arranged sequentially between the valve and the second switching module, and the position of the cell at the front adjacent to the valve is determined; (A5) The extraction needle moves to the top of the foremost defective cell and then moves down; the extraction needle aspirates a single defective cell; (A6) Rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector.

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