Flow cytometer and method of operation thereof

Abstract

The application provides a flow cytometer and a working method thereof. The flow cytometer comprises a flow cell and a detection unit. The application also comprises the following steps: fixing one end of a plurality of curved elastic fibers to the inner wall of the flow cell and leaving the other end of the elastic fibers to be suspended; and surrounding the elastic fibers to form a channel allowing single cells to pass through one by one; and blocking the through hole of the single cells by the gap between the adjacent elastic fibers. The application has the advantages of good focusing effect and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to cell detection, in particular to a flow cytometer and a working method thereof. BACKGROUND

[0002] In a flow cytometer, laminar sheath liquid focusing and acoustic focusing are usually used to make particles and cells in the center area of the flow cell, and then the spectral signal and image information of the particles are obtained through laser excitation and imaging.

[0003] However, the current focusing technology interacts with particles / cells by sheath liquid fluid or acoustic waves, and the interaction force is not strong enough, and the particles / cells cannot be well limited in the center position. The fluctuation of the position of the particles / cells will cause the stability of the detection signal of the instrument to be poor.

[0004] In addition, the laminar sheath focusing will make the particles / cells pass through the detection area at high speed, and the high flow rate is not conducive to obtaining more reliable and accurate spectral signals and clear image information. The acoustic focusing is also limited by the size of the particles / cells, and is difficult to be applied to the use scene with large particle size span. SUMMARY

[0005] In order to solve the above problems in the prior art, the present application provides a flow cytometer.

[0006] The purpose of the present application is achieved by the following technical solutions:

[0007] The flow cytometer comprises a flow cell and a detection unit; the flow cytometer further comprises:

[0008] Elastic fibers, one end of a plurality of curved elastic fibers is fixed to the inner wall of the flow cell, and the other end is suspended, and the elastic fibers are surrounded by a channel allowing single cells to pass through one by one; the gap between adjacent elastic fibers blocks the through hole of the single cells.

[0009] The purpose of the present application is also to provide a working method of the flow cytometer, and the purpose of the present application is achieved by the following technical solutions:

[0010] According to the working method of the flow cytometer of the present application, the working method is:

[0011] The cells enter the flow cell, and the cells in flow are constrained in the channel, and the liquid flows between the elastic fibers;

[0012] The single cells pass through the channel one by one, and the detection unit detects the single cells.

[0013] Compared with the prior art, the present application has the beneficial effects of:

[0014] 1. Good focusing effect;

[0015] The curved elastic fibers form channels to confine the cells, so that the cells pass through the channels one by one, and the cell focusing effect is improved;

[0016] 2. The detection signal is stable;

[0017] The elastic fibers are uniformly arranged on the inner wall of the flow cell, and the central axis of the formed channel is collinear with the central axis of the flow cell, so that the laser in the detection unit can be accurately focused on the single cell, thereby obtaining accurate spectral signals and imaging information;

[0018] The cells do not need to pass at high speed, and the signal stability is improved. BRIEF DESCRIPTION OF DRAWINGS

[0019] The disclosure of the present application will become more apparent from the following description in conjunction with the accompanying drawings. It is readily understood by those skilled in the art that the drawings are merely intended to illustrate the technical solutions of the present application, and are not intended to limit the scope of protection of the present application. In the drawings:

[0020] Figure 1 is a structural schematic diagram of a flow cytometer according to an embodiment of the present application;

[0021] Figure 2 is another structural schematic diagram of a flow cytometer according to an embodiment of the present application. DETAILED DESCRIPTION

[0022] Figures 1-2 The following description describes optional embodiments of the present application to teach those skilled in the art how to implement and reproduce the present application. Some conventional aspects have been simplified or omitted in order to explain the technical solutions of the present application. Those skilled in the art should understand that variations or substitutions derived from these embodiments will be within the scope of the present application. Those skilled in the art should understand that the following features can be combined in various ways to form multiple variations of the present application. Therefore, the present application is not limited to the following optional embodiments, but is only limited by the claims and their equivalents.

[0023] Embodiment 1:

[0024] Figure 1 The structural diagram of the flow cytometer according to an embodiment of the present application is schematically shown in Figure 1 As shown in the figure, the flow cytometer comprises:

[0025] The flow cell 11 and the detection unit, which are prior art in the art;

[0026] Elastic fibers 31-32, as Figure 2As shown, one end of the plurality of curved elastic fibers 31-32 is fixed to the inner wall of the flow cell 11, and the other end is suspended, and the elastic fibers 31-32 are surrounded by a channel allowing single cells 21 to pass one by one; the gap between adjacent elastic fibers 31-32 blocks the through hole of the single cell 21.

[0027] In order to facilitate detection, further, the central axis of the channel and the central axis of the flow cell 11 are collinear, and the plurality of elastic fibers 31-32 are uniformly arranged on the inner wall of the flow cell 11.

[0028] In order to make the single cell 21 pass through the channel one by one, further, along the flow direction of the cell 21, the radius of the channel gradually decreases, and in the natural state, the minimum radius of the channel is smaller than the radius of the cell 21.

[0029] The working method of the flow cytometer of the embodiment of the application is:

[0030] The cell 21 enters the flow cell 11, and the cell 21 in flow is constrained in the channel, and the liquid flows between the elastic fibers 31-32;

[0031] The single cell 21 passes through the channel one by one, and the detection unit detects the single cell 21.

[0032] Embodiment 2:

[0033] The application example of the flow cytometer and the working method thereof according to Embodiment 1 of the application in the detection of planktonic algae.

[0034] The particle size range of planktonic algae spans a large range (0.5 μm-1000 μm), and the shapes of the algae cells are diverse, and some algae cells will gather into groups, fibers, branches, and discs. In water ecological monitoring, optical fiber mirrors are often used for qualitative classification and quantitative counting of algae, which is tedious and time-consuming and requires professional knowledge of algae personnel, which may introduce subjective errors and is not conducive to the standardized management of algae monitoring data. At present, precision instruments such as FlowCAM and Cytosense that can be used for online monitoring of algae realize automatic monitoring of algae by spectral method and imaging method, and have the advantages of high efficiency, automation, and standardization. However, FlowCAM flow cell does not use sheath liquid laminar focusing technology, and the movement speed of the algae cells is <10 mm / s, which is easy to image, but the unfocused algae cells are not necessarily in the focal plane, which is not conducive to clear imaging. Cytosense uses sheath liquid laminar focusing technology, although the algae cells fluctuate in a small range near the focal plane, but the movement speed of the algae cells reaches 2 m / s, which makes imaging difficult.

[0035] In this application example, as shown in Figures 1-2As shown, the circumference of the inner wall of the flow cell 11 allows the arrangement of a plurality of elastic fibers 31-32, the curved first group of elastic fibers 31 forms a cell channel in the natural state, along the transmission direction of the cells 21, the radius of the channel gradually decreases, and the minimum radius is smaller than the radius of the cells 21, so that the cells 21 are extruded from the channel one by one, ensuring that the single cells 21 pass through the detection area in sequence on the central axis of the flow cell 11, the elastic fibers 31-32 are metal wires; the second group of elastic fibers 32 is located between the adjacent first group of elastic fibers 31, and is used to block the channel of the cells, and the flow cell 11 is made of quartz.

[0036] The working method of the flow cytometer of the embodiment of the application is as follows:

[0037] The algal cell sample is pretreated to disperse the clustered algae into single cells or fibers, etc., and if the concentration of algae in the sample is too high, deionized water can be added to dilute it to a suitable concentration.

[0038] The pretreated algal cell sample is introduced into the flow cell 11, and water flows through the gap between the elastic fibers 31-32, and the algal cells are gradually guided to the center position by the first group of elastic fibers 31 and are constrained in the channel.

[0039] Under the extrusion of the flowing cells 21, the front single cells 21 are gradually extruded from the tail end of the channel by supporting the first group of elastic fibers 31 one by one, and the detection unit detects the single cells 21.

[0040] The above embodiment illustrates that the elastic fibers are metal wires, and of course can also be glass wires or plastic wires or other fibers, as long as they can provide elastic force to constrain the cells.

Claims

1. A flow cytometer comprising a flow cell and a detection unit; characterized in that, The flow cytometer further comprises: Elastic fibers, one end of a plurality of curved elastic fibers is fixed on the inner wall of the flow cell, the other end is suspended, the curved elastic fibers surround a channel allowing single cells to pass one by one, the radius of the channel gradually decreases along the direction of cell flow; in a natural state, the minimum radius of the channel is smaller than the radius of the cell; the gap between adjacent elastic fibers blocks the passage of single cells.

2. The flow cytometer of claim 1, wherein, The central axis of the channel and the central axis of the flow cell are collinear.

3. The flow cytometer of claim 1, wherein, The elastic fibers are plastic filaments, glass filaments or metal filaments.

4. The flow cytometer of claim 1, wherein, The plurality of elastic fibers are uniformly arranged on the inner wall of the flow cell.

5. The working method of the flow cytometer according to any one of claims 1-4, the working method being: Cells enter the flow cell, cells in flow are constrained in the channel, liquid flows between elastic fibers; Single cells pass through the channel one by one, and a detection unit detects single cells.

6. The method of operating a flow cytometer of claim 5, wherein, Cells are extruded from the channel.

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