CN112179831A - Device and method for detecting defective cells - Google Patents

Abstract

The invention provides a detection device and method for defective cells. The detection device for defective cells includes a cell detector; a cell screening unit screens cells with different diameters; and a first switching module is used for screening the cells with different diameters. selectively sent to the first channel; the first identification unit is used to identify whether the single cell in the first channel is defective; the second switching module switches the channel according to the result of the first identification unit; the identification result is a defective cell, so the The first channel is connected to the second channel, the identification result is a non-defective cell, and the first channel is connected to the third channel; the positioning module is used to determine the position of the defective cell in the second channel; the transfer unit is used to determine the position of the cell. Defective cells or extracts thereof are transferred to the cell analyzer. The invention has the advantages of accurate detection, automation and the like.

Description

Device and method for detecting defective cells

technical field

The present invention relates to cell analysis, in particular to a detection device and method for defective cells.

Background technique

Single-cell technology has developed rapidly in recent years. With the continuous development of single-cell sequencing, single-cell transcriptome and other technologies, human beings have an in-depth understanding of cell population heterogeneity from the level of DNA and RNA. At the protein level, flow cytometry has always been the most commonly used single-cell analysis method, but due to problems such as cross color, flow cytometry can usually only detect 6-10 proteins at the same time.

Mass cytometry (Mass Cytometry) is a flow technology that uses the principle of mass spectrometry to perform multi-parameter detection of single cells. It inherits the characteristics of high-speed analysis of traditional flow cytometer, and has the high-resolution ability of mass spectrometry detection. This fusion technology can analyze more than 40 kinds of cell parameters at the same time at the single-cell level, which greatly increases the flow cytometry analysis. The ability to assess complex cellular systems and processes is a new development in flow cytometry. It uses metal element labels (antibodies) to label or recognize signal molecules on the surface and interior of cells, then separate single cells by flow cytometry, and then observe the atomic mass spectrum of single cells by ICP-MS, and finally combine the atomic mass spectrum data. It is converted into signal molecular data on the cell surface and inside, and the obtained data is subjected to dimensionality reduction analysis by computer analysis software, so as to realize the fine observation of cell phenotype and signal network.

In mass flow cytometry, after the cell samples are separated into single cells by the flow chamber and the liquid flow system, they all need to be analyzed by mass spectrometry in sequence, which is not conducive to the rapid acquisition of the analysis results of the target cells, and the efficiency is low. Since an analysis is only for a single cell, there is a problem of weak signal for the analysis of a specific component in the cell.

SUMMARY OF THE INVENTION

In order to solve the deficiencies in the above-mentioned prior art solutions, the present invention provides a detection device for detecting accurate and automated defective cells.

The purpose of this invention is to realize through the following technical solutions:

A detection device for defective cells, the detection device for defective cells includes a cell detector; the detection device for defective cells further includes:

a cell screening unit, the cell screening unit screens cells of different diameters;

a first switching module, the first switching module is used to selectively send the screened cells of different diameters to the first channel;

a first identification unit, the first identification unit is used to identify whether the single cell in the first channel has a defect;

The second switching module, the second switching module switches channels according to the result of the first identification unit; the identification result is a defective cell, the first channel is connected to the second channel, and the identification result is a non-defective cell, the first channel Connect the third channel;

a positioning module, the positioning module is used to determine the position of the defective cell in the second channel;

a transfer unit for transferring defective cells or extracts thereof at the determined position to the cell detector.

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

A detection method for defective cells, the detection method for defective cells comprises the following steps:

(A1) The cell suspension enters the cell screening unit, and cells with different diameters R _i are screened, i=1, 2...n, n≥2, and n∈N;

(A2) By switching, select cells with diameter R _m to enter the first channel, 1≤m≤n, and m∈N;

(A3) identifying whether cells in the first channel are defective;

If the cell is defective, the switching module is controlled so that the first channel is connected to the second channel, and the step (A4) is entered;

If the cell is not defective, the switching module is controlled so that the first channel is connected to the third channel;

(A4) Obtain the position of the defective cell in the second channel by using the positioning module;

(A5) The extraction needle moves to the upper side of the position and moves down; the extraction needle sucks a single defective cell at the position;

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

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

1. Accurate detection;

Cells with different diameters are screened out by the cell screening unit, which improves the accuracy of cell detection;

Defective cells are accurately identified by the identification unit, which improves the detection accuracy of defective cells;

During the detection process, multiple extraction needles absorb multiple defective cells (one extraction needle only absorbs a single defective cell), and multiple defective cells are simultaneously detected by techniques such as mass spectrometry, such as simultaneous ionization, which improves the detection signal intensity, correspondingly Improve detection accuracy;

2. Automation;

Cell screening, identification, aspiration, transfer, and detection can be automated, improving detection efficiency.

Description of drawings

The disclosure of the present invention will become more easily understood with reference to the accompanying drawings. Those skilled in the art can easily understand that these drawings are only used to illustrate the technical solutions of the present invention, and are not intended to limit the protection scope of the present invention. In the picture:

FIG. 1 is a flowchart of a method for detecting defective cells according to an embodiment of the present invention.

Detailed ways

Figure 1 and the following description describe alternative embodiments of the invention to teach those skilled in the art how to implement and reproduce the invention. In order to teach the technical solutions of the present invention, some conventional aspects have been simplified or omitted. Those skilled in the art will appreciate that variations or substitutions from these embodiments will fall within the scope of the present invention. Those skilled in the art will appreciate that the following features can be combined in various ways to form various variations of the invention. Thus, the present invention is not limited to the alternative embodiments described below, but only by the claims and their equivalents.

Example 1:

A detection device for defective cells, the detection device for defective cells includes:

Cell detectors, such as mass spectrometers;

a cell screening unit, the cell screening unit screens cells of different diameters;

a first switching module, the first switching module is used to selectively send the screened cells of different diameters to the first channel;

a first identification unit, the first identification unit is used to identify whether the single cell in the first channel has a defect;

The second switching module, the second switching module switches channels according to the result of the first identification unit; 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, and the identification result is a non-defective cell Defective cells, the first channel communicates with the third channel;

a positioning module, the positioning module is used to determine the position of the defective cell in the second channel;

a transfer unit for transferring defective cells or extracts thereof at the determined position to the cell detector.

In order to accurately screen cells with different diameters, further, the cell screening unit includes:

At least two interception modules, at least two interception modules are arranged on top of each other, and the interception modules include grooves; from top to bottom, the width of the grooves of the interception modules becomes smaller and communicated; the input end of the first switching module Connect to the groove outlet of each entrapment module.

In order to improve the screening effect of cells with different diameters and drive the cells to move forward, further, the cell screening unit further includes:

A vibrator, the vibrator is connected to the at least two trapping modules.

In order to accurately identify the defective cells, further, the first identification unit includes:

a light source, the light source is arranged on the side of the first channel;

a detector, the detector receiving the light emitted by the light source and passing through the first channel to obtain an image of the cell;

an identification module, the identification module identifies whether the cell is defective according to the image.

In order to accurately locate the position of the defective cells in the second channel, further, the positioning module is arranged on the lower side of the second channel, obtains an image of the defective cells in the second channel, and identifies the defective cells location; or,

The positioning module employs a recess adapted to accommodate a single defective cell.

In order to transfer the defective cells in the second channel, further, the transfer unit includes:

one or more extraction needles for aspirating single cells or extracts thereof;

a driving module, which is used to drive the up and down movement of the extraction needle;

The rotation module is used for rotating the extraction needle, so that the outlet of the extraction needle corresponds to the inlet of the cell detector.

In order to further confirm that the cells in the second channel have defects, further, a light source is disposed between the extraction needles, the positioning module receives the light emitted by the light source and passes through the first channel, obtains an image of the cells, and Identify whether cells are defective.

In order to temporarily store the defective cells, further, the detection device for the defective cells further includes:

and a valve, which is arranged on the second channel between the second switching module and the positioning module.

FIG. 1 shows a flowchart of a method for detecting defective cells according to an embodiment of the present invention, that is, a working method of the detection device according to this embodiment. As shown in FIG. 1 , the method for detecting defective cells includes the following steps:

(A1) The cell suspension enters the cell screening unit, and cells with different diameters R _i are screened, i=1, 2...n, n≥2, and n∈N;

(A2) By switching, select cells with diameter R _m to enter the first channel, 1≤m≤n, and m∈N;

(A3) using the first identification unit to identify whether the cells in the first channel have defects, such as through cell imaging, image analysis (machine vision), etc.;

If the cell is defective, the switching module is controlled so that the first channel is connected to the second channel, and the step (A4) is entered;

If the cell is not defective, the switching module is controlled so that the first channel is connected to the third channel;

(A4) Obtain the position of the defective cell in the second channel by using the positioning module;

(A5) The extraction needle moves to the upper side of the position and moves down; the extraction needle sucks a single defective cell at the position: for example, a single extraction needle sucks a single defective cell, or multiple extraction needles suck a plurality of defective cells ;

(A6) Rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector, so as to analyze the substances in the defective cells.

In order to facilitate the extraction needle to accurately absorb the defective cells, further, the positioning method of the defective cells is as follows:

Close the valve on the second channel, the defective cells entering the second channel are blocked by the valve, and the defective cells accumulate and temporarily store;

After the single-defective cells in the suction position are taken away by the extraction needle, the valve is opened, and one or more defective cells temporarily stored pass through the valve and enter the suction position;

Repeat the above method, so that the temporarily stored defective cells enter the suction position in sequence;

Alternatively, the defective cells entering the second channel are blocked and temporarily stored in sequence; one or more defective cells in the front row are sucked by the extraction needle, and other temporarily stored defective cells move forward.

Example 2:

An application example of the apparatus and method for detecting defective cells according to Embodiment 1 of the present invention.

In this application example, the cell screening unit adopts three layers of interception modules stacked up and down. The interception module has linear grooves. From top to bottom, the width of each layer of grooves becomes smaller, and the top and bottom are connected, so that the diameter of the trapped cells is reduced. getting smaller and smaller; the ultrasonic vibrator is connected to the retention module; the first switching module adopts a multi-port valve, so that the first channel can selectively connect with any linear groove, that is, receive cells of various diameters;

The first identification unit adopts 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 methods to identify defective cells, such as cell imaging and normal cell image comparison, deviations. Larger ones are identified as defective cells;

The second switching module adopts a three-way valve, so that the second channel selectively connects the second channel and the third channel. Connect with the second channel, otherwise connect the first channel and the third channel;

The positioning module adopts a groove for accommodating a single defective cell, and is arranged in the second channel;

A plurality of extraction needles are arranged on the first rotating module; the first rotating module is arranged on the second rotating module, and the rotating shaft of the second rotating module is horizontal; the rotating shaft of the first rotating module is vertical and the second rotating module the shaft of the module;

The cell detector adopts mass spectrometer with ion inlet;

The first channel, the second channel and the third channel are all transparent pipes, wherein the upper end of the part of the second channel has an opening, which is convenient for the extraction needle to enter the second channel to absorb the defective cells.

The detection method for defective cells of the present embodiment includes the following steps:

(A1) The cell suspension enters the cell screening unit, the ultrasonic vibrator drives the trapping module to vibrate, the suspension flows from top to bottom, and three types of cells with different diameters are screened. The cell with the largest diameter is located in the groove of the top trapping module. , the cell with the smallest diameter is in the groove of the lowermost retention module;

(A2) Select the cell with the largest diameter (that is, the cell in the uppermost trapping module groove) to enter the first channel through automatic switching of the multi-way valve;

(A3) using the first identification unit to identify whether the cells in the first channel have defects, and automatically identify the defective cells through cell imaging and image analysis technology;

后自动控制三通阀，使得第一通道连通第二通道，缺陷细胞通过三通阀进入第二通道，并在时间t₀+t₁后切换三通阀，使得第一通道和第二通道间关闭，L为第一识别单元和三通阀间第一通道的距离，v为细胞移动的速度，t₁为缺陷细胞通过三通阀的时间，也即在常规时间内三通阀保持了第一通道和第三通道间的连通，仅在识别出缺陷细胞后，才利用三通阀实现第一通道和第二通道间的连通；并进入步骤(A4)；If the cell is defective, then at time

After that, the three-way valve is automatically controlled, so that the first channel is connected to the second channel, and the defective cells enter the second channel through the three-way valve, and the three-way valve is switched after time t ₀ +t ₁ , so that the gap between the first channel and the second channel is closed, L is the distance between the first identification unit and the first channel of the three-way valve, v is the speed of cell movement, t ₁ is the time for the defective cell to pass through the three-way valve, that is, the three-way valve maintains the first For the communication between the first channel and the third channel, only after the defective cells are identified, the three-way valve is used to realize the communication between the first channel and the second channel; and proceed to step (A4);

If the cells are not defective, the three-way valve is controlled to switch, so that the first channel is connected to the third channel, and the non-defective cells enter the third channel;

(A4) using the groove to obtain the position of the defective cell in the second channel;

(A5) The extraction needle moves to the upper side of the position (the rotation axis of the first rotation module is vertical at this time), and moves down, the extraction needle absorbs a single defective cell at the position, and then moves up; the defective cells are continuously Passing through the three-way valve into the groove of the second channel, the first rotating module rotates, so that different extraction needles absorb the defective cells in the groove;

(A6) After the cells with the largest diameter all pass through the first identification unit, the second rotation module rotates around the horizontal axis, so that the outlet of the extraction needle corresponds to the inlet of the cell detector;

The electrodes on each extraction needle are discharged, and multiple defective cells absorbed are ionized at the same time, and enter the inlet of the mass spectrometer for analysis, thereby analyzing the substances in the defective cells.

Example 3:

According to the application example of the detection device and method for defective cells in Embodiment 1 of the present invention, the differences from Embodiment 2 are:

A valve is arranged on the second channel, and the valve is located between the three-way valve and the positioning groove; the defective cells passing through the three-way valve are temporarily stored in the second channel between the three-way valve and the valve, and the cells are arranged in order, The position of the cells in the front row adjacent to the valve is determined, so that the valve indirectly functions as a positioning module; with the rotation of the first rotation module, different extraction needles continuously suck up the defective cells in the front (when the front defective cells are in the front). After being aspirated, the rear cells move forward).

Example 4:

According to the application example of the detection device and method for defective cells in Embodiment 1 of the present invention, the differences from Embodiment 2 are:

A light source is arranged between the extraction needles, and a detector is arranged on the lower side of the second channel to obtain an image of the defective cells in the second channel, and the image is processed by the positioning module to obtain the position of the defective cell in the second channel; the extraction needle is used to absorb the Defective cells at the location.

Example 5:

According to the application example of the detection device and method for defective cells in Embodiment 1 of the present invention, the differences from Embodiment 2 are:

1. A valve is set on the second channel, and the valve is located between the three-way valve and the positioning groove; the defective cells passing through the three-way valve are temporarily stored in the second channel between the three-way valve and the valve, and pass through the valve. And open and close, so that the temporarily stored defective cells pass through the valve one by one (that is, only one cell passes through the valve at a time), and enter the positioning groove one by one; with the rotation of the first rotation module, different extraction needles Aspirate a single defective cell in the groove over time.

2. Set up a second identification unit, which adopts the same structure as the first identification unit, wherein the light source is set between the extraction needles, and the detector is set at the lower side of the second channel; using the second identification unit, it is confirmed again that it is in the second channel Whether the defective cells in the positioning groove are defective, if it is verified again that it is defective, the extraction needle will be controlled to suck up, if it is verified that there is no defect, the second channel will be discharged and no longer detected.

Claims (10)

1. A detection device for defective cells, comprising a cell detector; characterized in that, the detection device for defective cells further comprises: a cell screening unit, the cell screening unit screens cells of different diameters; a first switching module, the first switching module is used to selectively send the screened cells of different diameters to the first channel; a first identification unit, the first identification unit is used to identify whether the single cell in the first channel has a defect; The second switching module, the second switching module switches channels according to the result of the first identification unit; the identification result is a defective cell, the first channel is connected to the second channel, and the identification result is a non-defective cell, the first channel Connect the third channel; a positioning module, the positioning module is used to determine the position of the defective cell in the second channel; a transfer unit for transferring defective cells or extracts thereof at the determined position to the cell detector. 2. The detection device for defective cells according to claim 1, wherein the cell screening unit comprises: At least two interception modules, at least two interception modules are arranged on top of each other, and the interception modules include grooves; from top to bottom, the width of the grooves of the interception modules becomes smaller and communicated; the input end of the first switching module Connect to the groove outlet of each entrapment module. 3. The detection device for defective cells according to claim 2, wherein the cell screening unit further comprises: A vibrator, the vibrator is connected to the at least two trapping modules. 4. The detection device for defective cells according to claim 1, wherein the first identification unit comprises: a light source, the light source is arranged on the side of the first channel; a detector, the detector receiving the light emitted by the light source and passing through the first channel to obtain an image of the cell; an identification module, the identification module identifies whether the cell is defective according to the image. 5 . The detection device for defective cells according to claim 1 , wherein the positioning module is arranged on the lower side of the second channel, obtains images of defective cells in the second channel, and identifies the defective cells in the second channel. 6 . the location of the defective cell; or, The positioning module employs a recess adapted to accommodate a single defective cell. 6. The detection device for defective cells according to claim 1, wherein the transfer unit comprises: one or more extraction needles for aspirating single cells or extracts thereof; a driving module, which is used to drive the up and down movement of the extraction needle; The rotation module is used for rotating the extraction needle, so that the outlet of the extraction needle corresponds to the inlet of the cell detector. 7 . The detection device for defective cells according to claim 6 , wherein a light source is arranged between the extraction needles, and the positioning module receives the light emitted by the light source and passed through the first channel to obtain cells. 8 . images and identify whether cells are defective. 8. The detection device for defective cells according to claim 1, wherein the detection device for defective cells further comprises: and a valve, which is arranged on the second channel between the second switching module and the positioning module. 9. A detection method for defective cells, the detection method for defective cells comprises the following steps: (A1) The cell suspension enters the cell screening unit, and cells with different diameters R _i are screened, i=1, 2...n, n≥2, and n∈N; (A2) By switching, select cells with diameter R _m to enter the first channel, 1≤m≤n, and m∈N; (A3) identifying whether the cells in the first channel are defective; If the cell is defective, the switching module is controlled so that the first channel is connected to the second channel, and the process goes to step (A4); If the cell is not defective, the switching module is controlled so that the first channel is connected to the third channel; (A4) Obtain the position of the defective cell in the second channel by using the positioning module; (A5) The extraction needle moves to the upper side of the position and moves down; the extraction needle sucks a single defective cell at the position; (A6) Rotate the extraction needle so that the outlet of the extraction needle corresponds to the inlet of the cell detector. 10. The method for detecting defective cells according to claim 9, wherein the positioning mode of the defective cells is: Close the valve on the second channel, the defective cells entering the second channel are blocked by the valve, and the defective cells accumulate and temporarily store; After the single-defective cells in the suction position are taken away by the extraction needle, the valve is opened, and one or more defective cells temporarily stored pass through the valve and enter the suction position; Repeat the above method, so that the temporarily stored defective cells enter the suction position in sequence; Alternatively, the defective cells entering the second channel are blocked and temporarily stored in sequence; one or more defective cells in the front row are sucked by the extraction needle, and other temporarily stored defective cells move forward.

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