Flow cytometer, optical signal collection lens and sample analysis equipment

Abstract

The embodiment of the invention provides a flow cytometer, an optical signal collection lens and sample analysis equipment, and relates to the technical field of flow cytometers. The flow cytometer comprises a liquid path system and an optical system, the liquid path system comprises a sampling needle, a sheath liquid bottle and a flowing chamber, and the sampling needle and the sheath liquid bottle are both communicated with the flowing chamber and are respectively configured to convey a sample and sheath liquid to the flowing chamber; the optical system comprises a light source and an optical signal collecting lens, the optical signal collecting lens comprises a fluid channel, and the fluid channel is communicated with the flowing chamber; wherein the liquid path system further comprises a negative pressure tank and a waste liquid pump, the waste liquid pump is communicated with the flow chamber, the negative pressure tank is arranged between the waste liquid pump and the flow chamber, and negative pressure is formed in the negative pressure tank through work of the waste liquid pump, so that a sample is loaded to the flow chamber from the sampling needle and enters the fluid channel. Compared with the prior art, the flow cytometer has the advantages that the sample loading mode is improved, the operation procedure is simplified, and the continuous sample loading is realized.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] The present disclosure claims the priority of Chinese patent application No. 202410879963.7 filed with the Chinese Patent Office on July 2, 2024, and entitled “A Light Signal Collection Lens and Sample Analysis Device”, the entire contents of which are incorporated by reference in the present disclosure. Technical Field

[0003] The present disclosure relates to the technical field of flow cytometers, and in particular to a flow cytometer, an optical signal collection lens, and a sample analysis device. Background Art

[0004] Flow cytometers are usually configured to analyze and count cells or particles in samples. Flow cytometers are mainly composed of a fluidic system, an optical system, and a data analysis system. As one of the important components of a flow cytometer, the fluidic system transports samples containing fluorescently stained cells or particles together with sheath fluid to the flow chamber, and uses the principle of fluid dynamics to focus the cells or particles so that the cells or particles pass through the flow chamber detection area in sequence. A stable sample flow is the guarantee for subsequent signal detection and is also one of the core technologies of the entire instrument.

[0005] Most of the mainstream flow cytometers on the market currently have an absolute counting function. Absolute volume counting is achieved by measuring the volume of the sample being tested, and then dividing the measured number of cells by the volume of the sample being tested to obtain the concentration information of the sample being tested, thereby achieving the purpose of absolute counting. The absolute volume counting solution is mainly achieved by a quantitative pump that can quantitatively transport the sample volume, such as a syringe pump, a plunger pump, a peristaltic pump, etc. Usually, the pipeline connection, sample suction, sample loading, and cleaning are relatively complicated, which will also lead to problems such as poor consistency of test results and high carryover contamination rate. Syringe pumps and plunger pumps cannot continuously load samples, and need to first suck the sample and then load the sample, and the operation procedure is complicated. Summary of the invention

[0006] The present disclosure provides a flow cytometer, an optical signal collection lens, and a sample analysis device, which can improve the sample loading method, simplify the operation procedure, and perform continuous sample loading.

[0007] The embodiments of the present disclosure may be implemented as follows:

[0008] The present disclosure provides a flow cytometer, comprising:

[0009] Fluid system and optical system;

[0010] The fluid path system comprises a sampling needle, a sheath fluid bottle and a flow chamber, wherein the sampling needle and the sheath fluid bottle are both in communication with the flow chamber and are respectively configured to transport a sample and a sheath fluid to the flow chamber;

[0011] The optical system comprises a light source and a light signal collecting lens, wherein the light signal collecting lens comprises a fluid channel, wherein the fluid channel is in communication with the flow chamber, wherein the light source is configured to illuminate a sample flowing from the flow chamber into the fluid channel, and wherein the light signal collecting lens is configured to collect and transmit a light beam generated by the sample after being illuminated;

[0012] Wherein, the liquid circuit system also includes a negative pressure tank and a waste liquid pump, the waste liquid pump is connected to the flow chamber, and the negative pressure tank is arranged between the waste liquid pump and the flow chamber. The waste liquid pump works to form a negative pressure in the negative pressure tank, so that the sample is loaded from the sampling needle into the flow chamber and enters the fluid channel.

[0013] Optionally, the waste liquid pump is controlled to maintain the pressure in the negative pressure tank dynamically constant at a target pressure value.

[0014] Optionally, the negative pressure tank includes a first pressure sensor, which is configured to obtain a real-time pressure value in the negative pressure tank, use the real-time pressure value as a feedback value, and use the target pressure value as a target value, and perform PID control on the pressure in the negative pressure tank through the waste liquid pump.

[0015] Optionally, the target pressure value is P1, wherein -60Kpa≤P1≤-10Kpa.

[0016] Optionally, the fluid path system further comprises a sheath fluid pump, wherein the sheath fluid pump is connected between the sheath fluid bottle and the flow chamber, and the sheath fluid supply is adjusted by controlling the sheath fluid pump to adjust the flow rate of the sample to the flow chamber.

[0017] Optionally, the liquid circuit system also includes a flow meter, which is arranged on the pipeline between the sampling needle and the flow chamber, and the flow meter is configured to obtain a real-time flow value of the sample flowing to the flow chamber, and use the real-time flow value as a feedback value and a set flow value as a target value to perform PID control on the flow of the sample entering the sampling needle through the sheath liquid pump.

[0018] Optionally, during the sample detection process, the set flow value is Q1, wherein 5ul / min≤Q1≤200ul / min.

[0019] Optionally, the negative pressure tank comprises a negative pressure tank body and a damping device, wherein the damping device is disposed on the negative pressure tank body, and the damping device and the negative pressure tank body are internally connected;

[0020] And / or, the negative pressure tank comprises a negative pressure tank body, the negative pressure tank body is provided with a first flow channel and a pressure relief port, one end of the first flow channel is connected to the pressure relief port, the other end of the first flow channel is configured to be connected to the negative pressure tank body, and the pressure relief port is configured to achieve rapid pressure relief of the negative pressure tank;

[0021] And / or, the negative pressure tank includes a negative pressure tank body and a control valve, the negative pressure tank body is provided with a first flow path channel and a pressure relief port, the control valve is arranged in the first flow path channel, and is configured to realize the connection and disconnection of the negative pressure tank body and the pressure relief port.

[0022] Optionally, the liquid circuit system further comprises a swab, the swab is provided with a cleaning channel, and the sampling needle is slidably matched with the cleaning channel;

[0023] Preferably, the cleaning channel is slidable relative to the sampling needle.

[0024] Optionally, the liquid circuit system further includes a swab pump, one end of the cleaning channel is connected to the swab pump, and the other end of the cleaning channel is connected to the sheath fluid bottle;

[0025] And / or, the liquid path system further includes a cleaning liquid pump and a cleaning liquid bottle, one end of the cleaning liquid pump is connected to the cleaning liquid bottle, and the other end of the cleaning liquid pump is connected to the flow chamber.

[0026] Optionally, the liquid circuit system further includes a waste liquid bottle, which is connected to the waste liquid pump and is configured to receive the waste liquid output from the negative pressure tank;

[0027] And / or, the liquid circuit system further comprises a sheath liquid pump, a degasser and a first pipeline, and the sheath liquid bottle, the sheath liquid pump, the degasser and the flow chamber are sequentially arranged in the first pipeline;

[0028] And / or, the fluid path system includes a sheath fluid pump and a depulsation device, one end of the depulsation device is connected to the sheath fluid pump, and the other end of the depulsation device is connected to the flow chamber.

[0029] Optionally, the optical signal collecting lens comprises a first transmission part, a second transmission part and a collecting part which are sequentially arranged at intervals;

[0030] The first transmission part is provided with the fluid channel, and the fluid channel is configured to transport samples. The sample generates a lateral light beam after being irradiated by the laser. A deflection surface is formed on the side of the first transmission part close to the second transmission part, and the deflection surface is configured to correct the spherical aberration of the lateral light beam.

[0031] Optionally, the deflection surface is wavy and concave;

[0032] And / or, the first transmission part further includes a reflection surface, the reflection surface is a curved surface and is located on a side of the fluid channel away from the second transmission part;

[0033] And / or, the first transmission part further includes a reflective surface and a refractive surface, the refractive surface is a plane and is located between the reflective surface and the fluid channel;

[0034] And / or, the first transmission part further includes a reflective surface, the reflective surface is coated with a reflective film, and the reflective film is a dielectric film or a metal film.

[0035] The cross section of the fluid channel is rectangular, and the lateral edge of the reflective surface is located on the extension line of the diagonal line of the same rectangular cross section as the fluid channel;

[0036] Preferably, the side length of the rectangular cross section of the fluid channel is between 100 um and 500 um.

[0037] The embodiment of the present disclosure further provides an optical signal collecting lens, which includes:

[0038] A first transmission part, a second transmission part and a collection part are arranged in sequence and at intervals;

[0039] The first transmission part has a fluid channel, which is configured to transport a sample. The sample generates a lateral light beam after being irradiated by a laser. A deflection surface is formed on one side of the first transmission part close to the second transmission part, and the deflection surface is configured to correct spherical aberration of the lateral light beam.

[0040] Optionally, the deflecting surface is wavy and concave.

[0041] Optionally, the first transmission part further includes a reflection surface, which is a curved surface and is located on a side of the fluid channel away from the second transmission part.

[0042] Optionally, the first transmission portion further comprises a refractive surface, which is a plane and is located between the reflective surface and the fluid channel;

[0043] And / or, the reflective surface is coated with a reflective film, and the reflective film is a dielectric film or a metal film.

[0044] Optionally, the second transmission part is a triplet lens, and includes a first sub-lens, a second sub-lens and a third sub-lens which are sequentially arranged along a direction from the collecting part to the first transmission part.

[0045] Optionally, the first sub-lens is arc-shaped, and a middle position of the first sub-lens is closer to the collecting portion than to both ends;

[0046] And / or, the third sub-lens is arc-shaped, and a middle position of the third sub-lens is closer to the first transmission portion relative to both ends.

[0047] An embodiment of the present disclosure further provides a sample analysis device, which includes the flow cytometer or the optical signal analysis lens.

[0048] The beneficial effects of the flow cytometer of the embodiment of the present disclosure include, for example:

[0049] The flow cytometer comprises a liquid circuit system and an optical system; the liquid circuit system comprises a sampling needle, a sheath liquid bottle and a flow chamber, the sampling needle and the sheath liquid bottle are both connected to the flow chamber and are respectively configured to transport samples and sheath liquid to the flow chamber; the optical system comprises a light source and a light signal collecting lens, the light signal collecting lens comprises a fluid channel, the fluid channel is connected to the flow chamber, the light source is configured to irradiate the sample flowing from the flow chamber into the fluid channel, and the light signal collecting lens is configured to collect and transmit the light beam generated by the sample after being irradiated; wherein, the liquid circuit system also comprises a negative pressure tank and a waste liquid pump, the waste liquid pump is connected to the flow chamber, the negative pressure tank is arranged between the waste liquid pump and the flow chamber, the waste liquid pump works to form a negative pressure in the negative pressure tank, so that the sample is loaded from the sampling needle into the flow chamber and enters the fluid channel. During operation, the liquid circuit system can work through the waste liquid pump to form negative pressure in the negative pressure tank. Through the connection between the negative pressure tank and the flow chamber, the negative pressure causes the sample to be loaded from the sampling needle to the flow chamber. The sheath liquid bottle is configured to provide sheath liquid to the flow chamber to wrap the sample to form a laminar flow. The light source irradiates the sample wrapped by the sheath liquid and generates a light beam. The light signal collection lens is configured to transmit and collect the light beam. Compared with the prior art, the flow cytometer has improved the loading method, simplified the operation procedure, and realized continuous loading. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present disclosure and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0051] Figure 1 It is a schematic diagram of the structure of the liquid circuit system provided in the embodiment of the present disclosure;

[0052] Figure 2 A schematic diagram of a cleaning channel of a swab provided in an embodiment of the present disclosure;

[0053] Figure 3 A schematic diagram of the structure of an optical signal collection lens provided in an embodiment of the present disclosure;

[0054] Figure 4A schematic diagram of the operation of the optical signal collecting lens provided in an embodiment of the present disclosure;

[0055] Figure 5 A schematic diagram of the structure of an optical signal collection lens provided in an embodiment of the present disclosure;

[0056] Figure 6 A schematic diagram of the operation of the optical system provided in an embodiment of the present disclosure;

[0057] Figure 7 A schematic cross-sectional view of a flow chamber provided in an embodiment of the present disclosure;

[0058] Figure 8 This is a control schematic diagram provided in an embodiment of the present disclosure.

[0059] Icons: 100-negative pressure tank; 110-control valve; 111-first valve; 112-second valve; 113-third valve; 114-fourth valve; 115-fifth valve; 120-negative pressure tank body; 121-pressure relief port; 122-first flow channel; 123-second flow channel; 130-damping device; 140-first pressure sensor; 190-first pipeline; 200-sheath pump; 240-light source; 245-forward light beam; 246-side light beam; 250-light signal collection lens; 260-first transmission part; 261-fluid channel; 262-baffle surface; 263-refractive surface; 264-reflective surface; 265-reflective film; 270-second transmission part; 271-first sub-lens; 272-second sub-lens; 273-third sub-lens; 280-collecting part; 300-flow chamber; 310-flow cell; 350-sample delivery needle; 400-sampling needle; 500-waste liquid pump; 610-swab; 611-cleaning channel; 6111-first sub-channel; 6112-second sub-channel; 620-swab pump; 700-sheath liquid bottle; 800-degasser; 900-first filter; 10-depulsation device; 20-cleaning liquid pump; 30-cleaning liquid bottle; 40-second pressure sensor; 50-waste liquid bottle; 60-sixth valve; 70-flow meter; 80-check valve; 90-intake filter; 1100-liquid path system; 1200-optical system; 2000-sample. DETAILED DESCRIPTION

[0060] In order to make the purpose, technical solutions and advantages of the embodiments of the present disclosure clearer, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below in conjunction with the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all the embodiments. The components of the embodiments of the present disclosure described and shown in the drawings here can be arranged and designed in various different configurations.

[0061] Therefore, the following detailed description of the embodiments of the present disclosure provided in the accompanying drawings is not intended to limit the scope of the present disclosure claimed for protection, but merely represents selected embodiments of the present disclosure. Based on the embodiments in the present disclosure, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present disclosure.

[0062] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0063] In the description of the present disclosure, it should be noted that if the terms "upper", "lower", "inside", "outside", etc. appear, the orientation or position relationship indicated is based on the orientation or position relationship shown in the drawings, or is the orientation or position relationship in which the product of the invention is usually placed when used. It is only for the convenience of describing the present disclosure and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present disclosure.

[0064] In addition, the terms “first”, “second”, etc., if used, are merely used to distinguish between the descriptions and should not be understood as indicating or implying relative importance.

[0065] It should be noted that, in the absence of conflict, the features in the embodiments of the present disclosure may be combined with each other.

[0066] As mentioned in the background technology, flow cytometers are usually configured to analyze and count cells or particles in samples. Flow cytometers are mainly composed of a fluid system, an optical system, and a data analysis system. As one of the important components of a flow cytometer, the fluid system transports samples containing cells or particles together with sheath fluid to the flow chamber, focuses the cells or particles using the principle of fluid dynamics, and allows the cells or particles to pass through the flow chamber detection area in sequence. A stable sample flow is the guarantee for subsequent signal detection and is also one of the core technologies of the entire instrument.

[0067] Most of the mainstream flow cytometers on the market currently have an absolute counting function. Absolute volume counting is achieved by measuring the volume of the sample being tested, and then dividing the measured number of cells by the volume of the sample being tested to obtain the concentration information of the sample being tested, thereby achieving the purpose of absolute counting. The absolute volume counting solution is mainly achieved by a quantitative pump that can quantitatively transport the sample volume, such as a syringe pump, a plunger pump, a peristaltic pump, etc. Usually, the pipeline connection, sample suction, sample loading, and cleaning are relatively complicated, which will also lead to problems such as poor consistency of test results and high carryover contamination rate. Syringe pumps and plunger pumps cannot continuously load samples, and need to first suck the sample and then load the sample, and the operation procedure is complicated.

[0068] In view of this, please refer to Figure 1 and Figure 2 The flow cytometer and control method thereof provided in the embodiments of the present disclosure can solve this problem, which will be described in detail below.

[0069] In this embodiment, a flow cytometer is provided, including a liquid system 1100 and an optical system 1200, wherein the liquid system 1100 is mainly configured to transmit liquids such as a sample 2000, a sheath liquid, a cleaning liquid, and a waste liquid, and the optical system 1200 is configured to irradiate the sample 2000 with a light source to form a specific light beam, and then transmit and collect the light beam through an optical signal collection lens 250. In addition, the light source can specifically be a laser 240.

[0070] Next, please refer to Figure 1 First, the liquid circuit system 1100 in this embodiment is specifically introduced. The liquid circuit system 1100 includes a sampling needle 400, a flow chamber 300, a sheath liquid pump 200, a negative pressure tank 100, a waste liquid pump 500, a sheath liquid bottle 700 and other components. The sampling needle 400, the sheath liquid bottle 700 and the negative pressure tank 100 are all connected to the flow chamber 300. The sampling needle 400 and the sheath liquid bottle 700 are respectively configured to transport the sample 2000 and the sheath liquid to the flow chamber 300. The sheath liquid pump 200 is connected between the sheath liquid bottle 700 and the flow chamber 300 and is configured to provide power for sheath liquid transmission. The waste liquid pump 500 is connected to the end of the negative pressure tank 100 away from the flow chamber 300, and is configured to suck the inner cavity of the negative pressure tank 100 to create a negative pressure environment, so that the negative pressure tank 100 can provide negative pressure to the flow chamber 300. The flow chamber 300 is configured to accommodate the sample 2000 and the sheath fluid for detection by cooperating with the optical system 1200 .

[0071] refer to Figure 1 and Figure 7 In order to facilitate the collection of waste liquid, the liquid path system 1100 also includes a waste liquid bottle 50. In addition, the liquid path system 1100 also includes a sample delivery needle 350. The sample delivery needle 350 is located in the flow chamber 300 and is connected to the sampling needle 400.

[0072] It is worth noting that during the operation, the waste liquid pump 500 can be controlled to maintain the pressure in the negative pressure tank 100 dynamically constant at the target pressure value, and then the flow rate of the sample 2000 can be adjusted by adjusting the sheath liquid flow rate, rather than dynamically maintaining the pressure difference at both ends of the flow chamber 300 stable.

[0073] Furthermore, the sheath liquid flow rate can be changed directly by changing the rotation speed of the sheath liquid pump 200, thereby changing the sample loading speed. There is no need to set a step flow valve to control the sheath liquid flow rate, and a larger flow adjustment range can be obtained.

[0074] The negative pressure tank 100 includes a negative pressure tank body 120 and multiple control valves 110. The control valve 110 can be a solenoid valve. The multiple control valves are arranged on the negative pressure tank body 120. In the present embodiment, the number of the control valves 110 is five, and the five control valves 110 are respectively a first valve 111, a second valve 112, a third valve 113, a fourth valve 114 and a fifth valve 115.

[0075] It is worth noting that a plurality of control valves 110 can be arranged on the integrated cover of the negative pressure tank 100. Through the intelligent operation of each control valve 110, the degree of automation can be improved, and it has the advantages of high integration and small size.

[0076] The first valve 111 is configured to control the conduction or blocking between the waste pump 500 and the flow chamber 300, so that when the first valve 111 is opened, the conduction between the waste pump 500 and the flow chamber 300 is convenient for implementing the sample loading process.

[0077] It should be noted that in order to facilitate the negative pressure tank 100 to achieve the function of rapid pressure relief, the negative pressure tank body 120 is provided with a first flow path channel 122 and a pressure relief port 121, one end of the first flow path channel 122 is connected to the pressure relief port 121, and the other end of the first flow path channel 122 is configured to be connected to the inside of the negative pressure tank body 120.

[0078] The third valve 113 is disposed in the first flow channel 122, and the third valve 113 is configured to control the conduction or blocking of the first flow channel 122 to achieve the conduction or blocking of the pressure relief port 121 and the negative pressure chamber inside the negative pressure tank body 120, wherein the negative pressure tank 100 also includes an air intake filter 90, and the pressure relief port 121 and the air intake filter 90 are connected.

[0079] In addition, in order to facilitate the monitoring of the pressure inside the negative pressure tank 100 and at the same time, accurately control the pressure inside the negative pressure tank 100 and reduce pressure fluctuations, the negative pressure tank 100 also includes a first pressure sensor 140 and a damping device 130. The negative pressure tank body 120 is provided with a second flow path channel 123. One end of the second flow path channel 123 is connected to the negative pressure tank body 120, and the first pressure sensor 140 is connected to the other end of the second flow path channel 123. The first pressure sensor 140 can be connected to the negative pressure chamber inside the negative pressure tank body 120 through the second flow path channel 123. The first pressure sensor 140 is convenient for monitoring the pressure inside the negative pressure tank 100.

[0080] When working, the first pressure sensor 140 is configured to obtain the real-time pressure value in the negative pressure tank 100, take the real-time pressure value as the feedback value, take the target pressure value as the target value, and perform PID control on the pressure in the negative pressure tank 100 through the waste liquid pump 500. In this process, the target value and the feedback value belong to the same attribute value, and there is no need for indirect conversion, so the control is simpler, the real-time feedback is faster, and the structure is simpler.

[0081] The damping device 130 is disposed in the negative pressure tank body 120 , and the damping device 130 is communicated with the negative pressure tank body 120 , that is, the damping device 130 is communicated with the negative pressure chamber inside the negative pressure tank body 120 .

[0082] The damping device 130 can be an air intake damping tube, or a throttle valve, a speed regulating valve or a filter, etc. The damping device 130 is connected to the atmosphere, which is equivalent to a pressure stabilizing device or a buffer, etc. The damping device 130 facilitates precise control of the pressure in the negative pressure tank 100 and effectively reduces pressure fluctuations.

[0083] It is worth noting that when the pressure in the negative pressure tank 100 fluctuates, especially when the pressure fluctuation caused by the fluctuation of the output of the waste liquid pump 500 is too large, the air intake of the damping device 130 will also change accordingly, thereby effectively reducing the pressure fluctuation in the negative pressure tank 100. The damping device 130 serves as a bypass of the negative pressure output end, which can make the pressure fluctuation at the negative pressure output end more balanced, thereby providing a guarantee for the stable detection of subsequent signals.

[0084] In addition, it should be noted that the negative pressure tank 100 can realize real-time emptying of the liquid in the negative pressure tank 100 without storing waste liquid, and therefore there is no need to set up a liquid level sensor.

[0085] Therefore, the waste liquid pump 500 in this embodiment can use a diaphragm pump with lower precision and lower cost, and can also accurately control the pressure in the negative pressure tank 100 to achieve an effect of smaller output pressure fluctuations.

[0086] Please refer to Figure 2 Combined with Figure 1 The liquid path system 1100 also includes a swab 610, a swab pump 620 and a sheath fluid bottle 700. The swab 610 is provided with a cleaning channel 611. The swab pump 620 is connected to one end of the cleaning channel 611, and the other end of the cleaning channel 611 is connected to the sheath fluid bottle 700. The sheath fluid pump 200 is arranged on the pipeline between the sheath fluid bottle 700 and the other end of the cleaning channel 611.

[0087] Among them, the sampling needle 400 and the cleaning channel 611 can slide relative to each other to facilitate the cleaning of the sampling needle 400. Here, the relative sliding of the sampling needle 400 and the cleaning channel 611 can be understood as the sampling needle 400 does not move, while the swab 610 moves downward, or the swab 610 keeps its position unchanged, while the sampling needle 400 moves upward, which is equivalent to the retraction process of the sampling needle 400.

[0088] In this embodiment, it is preferred that the sampling needle 400 remains stationary while the swab 610 moves to perform the cleaning operation, and the configuration is to shorten the extended length of the sampling channel as much as possible to facilitate the reduction of the fixed volume.

[0089] Among them, Figure 2 As shown, the cleaning channel 611 has a main channel and a first sub-channel 6111 and a second sub-channel 6112 connected to the main channel. The sampling needle 400 and the main channel of the cleaning channel 611 can slide relative to each other. The first sub-channel 6111 and the swab pump 620 are connected through a pipeline. The liquid circuit also includes a sixth valve 60 (a solenoid valve can be selected). The sixth valve 60 is arranged in the pipeline between the first sub-channel 6111 and the swab pump 620. The second sub-channel 6112 is connected to the sheath liquid pump 200. The fourth valve 114 is configured to simultaneously connect the sheath liquid pump 200 and the second sub-channel 6112, so as to control the conduction or blocking of the pipeline between the sheath liquid pump 200 and the second sub-channel 6112.

[0090] In addition, in this embodiment, the liquid path system 1100 also includes a degasser 800 and a first pipeline 190. The sheath fluid bottle 700, the sheath fluid pump 200, the degasser 800 and the flow chamber 300 are sequentially arranged in the first pipeline 190. The degasser 800 can remove air from the sheath fluid, solve the influence of bubbles on the liquid flow, improve the stability of the liquid flow, improve the detection accuracy, and at the same time expand the adaptability of the flow cytometer to different usage environments.

[0091] Specifically, the liquid circuit system 1100 further includes a first filter 900 and a depulsation device 10, and the sheath liquid bottle 700, the sheath liquid pump 200, the depulsation device 10, the first filter 900, the degasser 800 and the flow chamber 300 are sequentially arranged in the first pipeline 190. During operation, the main function of the depulsation device 10 is to eliminate the pulsation generated when the sheath liquid pump 200 is working, thereby ensuring the transmission stability of the liquid circuit system.

[0092] The fifth valve 115 is configured to connect the degasser 800 and the flow chamber 300 at the same time. The fifth valve 115 is also disposed on the first pipeline 190 . The fifth valve 115 can control the conduction or blocking of the first pipeline 190 .

[0093] The fourth valve 114 is configured to simultaneously connect the first filter 900 and the second sub-channel 6112 , so as to control the conduction or blocking of the pipeline between the sheath liquid pump 200 and the second sub-channel 6112 .

[0094] At the same time, the liquid circuit system 1100 also includes a second filter (not shown), which can be arranged in the first pipeline 190 between the sheath liquid pump 200 and the sheath liquid bottle 700. The sheath liquid pump 200 can use a high-precision ceramic pump, and the second filter can protect the sheath liquid pump 200.

[0095] It should be noted that, in some embodiments, the sheath liquid bottle 700 , the sheath liquid pump 200 , the first filter 900 , the depulsation device 10 , the degasser 800 and the flow chamber 300 may be sequentially disposed in the first pipeline 190 .

[0096] In some embodiments, the fluid system 1100 may include the second filter but not the first filter 900 and the depulsation device 10 .

[0097] Alternatively, the liquid path system 1100 includes the first filter 900 and the depulsation device 10, but no second filter is provided. In this case, the sheath liquid pump 200 can be a peristaltic pump.

[0098] In order to facilitate the cleaning process of the liquid circuit, the liquid circuit system 1100 also includes a cleaning liquid pump 20 and a cleaning liquid bottle 30. One end of the cleaning liquid pump 20 (which can be understood as the liquid inlet end) is connected to the cleaning liquid bottle 30, and the other end of the cleaning liquid pump 20 (which can be understood as the liquid outlet end) is connected to the flow chamber 300.

[0099] The cleaning liquid pump 20 can transport the cleaning liquid in the cleaning liquid bottle 30 to the flow chamber 300. At the same time, the cleaning liquid can enter the sample delivery needle, flow through the flowmeter 70 and the sampling needle 400, and complete the cleaning of the sampling channel and the flow chamber 300. One path of waste liquid can enter the waste liquid bottle 50 through the first valve 111, the negative pressure tank 100 and the waste liquid pump 500, and the other path of waste liquid can enter the waste liquid bottle 50 through the sampling needle 400, the first sub-channel 6111 and the swab pump 620.

[0100] In addition, the liquid circuit system 1100 also includes a second pressure sensor 40 and a flow meter 70. The flow meter 70 is arranged on the pipeline between the sampling needle 400 and the flow chamber 300. Specifically, the flow meter 70 can be arranged on the pipeline between the sampling needle 400 and the sample delivery needle, and the second pressure sensor 40 is arranged on the pipeline between the flow chamber 300 and the sheath liquid pump 200.

[0101] In addition, the liquid circuit system 1100 also includes a one-way valve 80. The degasser 800 is connected to the pipeline between the cleaning channel 611 of the swab 610 and the swab pump 620 through a third pipeline. The one-way valve 80 is arranged in the third pipeline. The one-way valve 80 can prevent backflow.

[0102] In addition, in the present embodiment, the second valve 112 is configured to simultaneously connect the pipeline between the sixth valve 60 and the swab pump 620 and the flow chamber 300 to control the second valve 112 to simultaneously connect the pipeline between the sixth valve 60 and the swab pump 620 and the pipeline between the flow chambers 300 to be turned on or off. In this way, when the third valve 113 and the first valve 111 are opened, outside air can enter the negative pressure tank 100, so that the liquid in the flow chamber 300 can flow through the second valve 112, the swab pump 620 and the waste liquid bottle 50 in sequence, so as to achieve the purpose of emptying the flow chamber 300.

[0103] refer to Figure 3-Figure 6 , an embodiment of the present disclosure provides an optical signal collecting lens 250, which is configured as a flow cytometer or a sample analysis device, and the optical signal collecting lens 250 includes a first transmission part 260, a second transmission part 270 and a collection part 280 which are sequentially arranged at intervals;

[0104] The first transmission part 260 is provided with a fluid channel 261, the fluid channel 261 is configured to transport the sample 2000, the sample 2000 is configured to receive the irradiation of the laser and generate a divergent light beam, and a deflection surface 262 is formed on one side of the first transmission part 260 close to the second transmission part 270, and the deflection surface 262 is configured to correct the spherical aberration of the divergent light beam. It should be noted that the laser can also be understood as one of the light sources 240.

[0105] During operation, the optical signal collecting lens 250 can transport the sample 2000 through the fluid channel 261, and make the sample 2000 generate a divergent light beam while receiving laser irradiation, and then the divergent light beam is transmitted from the first transmission part 260 to the second transmission part 270 and the collecting part 280, and the deflection surface 262 is configured to correct the spherical aberration of the light beam transmitted by the first transmission part 260, thereby improving the light beam transmission and collection effect. It is worth noting that the first transmission part 260 can be understood as Figure 7 The flow cell 310 is disposed on the flow chamber 300 , and the fluid channel in the flow cell 310 can also be regarded as the fluid channel 261 in the first transmission part 260 .

[0106] Specifically, the baffle surface 262 is wavy and concave, and the wavy means that the surface of the baffle surface 262 is an arc with inconsistent curvature, which can also be understood as different positions on the baffle surface 262 have ups and downs; the concave means that the two ends of the baffle surface 262 are closer to the second transmission part 270 than the middle position, and the two ends of the baffle surface 262 refer to the two ends of the baffle surface 262 in the extension direction of the fluid channel 261. The optical signal collection lens 250 limits the shape of the baffle surface 262, so that the transmission of the light beam passing through the baffle surface 262 is more efficient, which is reflected in the better collection effect on the subsequent collection part 280, thereby achieving the maximum efficiency of collecting the lateral light beam 246.

[0107] It is worth noting that the fluid channel 261 is linear, and the extension direction of the fluid channel 261 is perpendicular to the direction from the first transmission part 260 to the second transmission part 270 to the collection part 280. In addition, the light source 240 irradiates the sample 2000 to generate a forward light beam 245 and a side light beam 246 at the same time, and the optical signal collection lens 250 is configured to collect the side light beam 246.

[0108] In addition, in order to make the surface shape of the deflector surface 262 more accurately determined, the surface shape calculation equation of the deflector surface 262 can be limited to: z(h)=h2 / R{1+[1-(1+k)h2 / R2]1 / 2}+A2h2+A4h4+A6h6+A8h8+A10h10+…+Anhn. The specific position of each point on the deflector surface 262 can be determined by this calculation formula, which can improve the processing convenience and facilitate repeated processing.

[0109] Among them, h is the distance from each point on the deflection surface 262 to the optical axis, R is the radius of curvature, k is the quadratic surface constant, and A2, A4, A6, A8, A10...An are higher-order surface coefficients.

[0110] Specifically, the actual distance may be calculated according to the formula h=(x2+y2)1 / 2, where x and y are the relative position coordinates of each point.

[0111] Moreover, the specific values ​​of each parameter can be obtained by inferring from the obtained optical signal effect according to multiple actual experiments. Preferably, R=-31.851, k=0, A2=0, A4=1.041-3, A6=8.762-5, A8=-4.93-6, A10=3.103-7. It should be noted that when the n value in An is larger, the specific parameter is smaller. Therefore, when n reaches a certain value, its influence on the final calculated value of z(h) is quite limited. Under conventional calculation conditions, n=10 can be taken, that is, the preferred surface shape calculation equation for the baffle 262 is: z(h)=h2 / R{1+[1-(1+k)h2 / R2]1 / 2}+A2h2+A4h4+A6h6+A8h8+A10h10.

[0112] refer to Figure 3-Figure 5 The first transmission portion 260 further includes a refractive surface 263 , which is a plane and is located on a side of the fluid channel 261 away from the second transmission portion 270 .

[0113] When a divergent light beam is generated, the divergent light beam first passes through the side wall of the fluid channel 261 and then is transmitted to the refractive surface 263 . After being refracted by the refractive surface 263 , the divergent light beam continues to be transmitted in a direction away from the fluid channel 261 .

[0114] refer to Figure 3-Figure 5 The first transmission part 260 further includes a reflective surface 264 , which is a curved surface and is located on a side of the refractive surface 263 away from the second transmission part 270 . That is, the refractive surface 263 is located between the reflective surface 264 and the fluid channel 261 .

[0115] During operation, the reflective surface 264 is configured to receive the light beam transmitted through the refractive surface 263 and then reflect it back to the refractive surface 263 to move it toward the direction close to the second transmission portion 270 , thereby reflecting the light beam.

[0116] Specifically, in order to improve the reflection effect, the surface of the reflective surface 264 can also be coated with a reflective film 265, which can be a dielectric film or a metal film. Among them, the dielectric film can be composed of multiple layers of oxide materials with high and low refractive indices, and the overall material thickness is about 0.1nm to 10nm, and there is no limitation on its specific thickness. After passing through the multi-layer structure, the light beam is folded back multiple times, thereby realizing the reflection of most of the light beam, thereby improving the reflectivity, so that its actual reflectivity can reach more than 99%. Although the reflectivity is also limited by the impact of the incident angle, by setting a refractive surface 263 on the side of the reflective surface 264 close to the fluid channel 261, the angle of the light beam incident on the reflective surface 264 can be adjusted to improve the collection efficiency of the light beam after reflection by the reflective surface 264 and reduce light loss. Metal films are generally composed of metals such as gold, silver, and aluminum materials. Different from the inconsistent reflectivity of individual metal materials in different bands, in the visible light band, considering the cost and stability, silver film or aluminum film is preferred, and the reflectivity can reach more than 90%.

[0117] In this embodiment, the cross section of the fluid channel 261 is rectangular, and the lateral edge of the reflecting surface 264 is located on the extension line of the diagonal of the same rectangular cross section as the fluid channel 261, and the side length of the rectangular cross section of the fluid channel 261 is preferably between 100um and 500um. Among them, the side length of the rectangular cross section includes the length L along the irradiation direction of the light source 240 and the width D perpendicular to the irradiation direction of the light source 240, and the range of both is between 100um and 500um. The lower the lower limit of the side length, the narrower the channel, which will make the flow resistance too large, resulting in a larger pressure difference at both ends, and then the negative pressure of the negative pressure tank 100 is too large; the higher the upper limit of the side length, the wider the channel, which is not conducive to compressing the core flow, and will result in excessive sheath liquid flow, and excessive sheath liquid consumption will also cause negative effects such as increased costs. This solution limits the upper and lower limits of the side length within a reasonable range, so that the channel width can be moderate, the flow resistance will not be too large, the pressure difference at both ends is reasonable, the negative pressure of the negative pressure tank 100 is reasonable, and it is conducive to compressing the core flow and avoiding excessive sheath liquid flow and increasing costs.

[0118] refer to Figure 3 and Figure 5 , the second transmission part 270 is a convex lens.

[0119] In this embodiment, the second transmission portion 270 is a triplet lens, and includes a first sub-lens 271 , a second sub-lens 272 , and a third sub-lens 273 which are sequentially arranged along a direction from the collecting portion 280 to the first transmission portion 260 .

[0120] Of course, optionally, the second transmission part 270 can also be a single lens, a doublet lens, a quadruplet lens, a quintuplet lens, etc., and there is no limitation on the specific number of sub-lenses therein.

[0121] refer to Figure 3-Figure 5 The first sub-lens 271 is arc-shaped, and the middle position of the first sub-lens 271 is closer to the collecting portion 280 relative to the two ends; and the third sub-lens 273 can also be arc-shaped, and the middle position of the third sub-lens 273 is closer to the first transmission portion 260 relative to the two ends; and the second sub-lens 272 is a convex lens, and the two sides of the second sub-lens 272 are glued to the first sub-lens 271 and the third sub-lens 273 respectively.

[0122] During the propagation of the light beam, after passing through the deflection surface 262, the light beam will enter the air portion between the first transmission portion 260 and the second transmission portion 270, and then reach the third sub-lens 273 and successively pass through the second sub-lens 272 and the first sub-lens 271 before being emitted, and then converge to the collection portion 280 to form a light spot with concentrated energy.

[0123] It is worth noting that the collecting part 280 generally places detectors, such as silicon-based photodetectors such as CCD, PD, APD or PMT, and may also place optical elements with other functions, including but not limited to apertures, pinholes or optical fiber bundles, etc., configured to control the transmitted light.

[0124] In addition, the optical signal collecting lens 250 adopts a folding structure, which adds a reflector system to the traditional refractive material. Since the chromatic aberration correction of the refractive material in a wide band is relatively difficult, even if the combination of crown glass and flint glass is continuously increased, the remaining secondary spectral chromatic aberration is relatively large, while the pure reflection system has no chromatic aberration in any desired bandwidth range, and it also promotes a simple structure and a reduction in the number of lenses. Therefore, by adding a reflector system to the traditional refractive material, the difficulty of chromatic aberration correction can be reduced, and the lens structure can be simplified.

[0125] Since the fluorescence signal excited by flow cytometry is a wide spectrum from 400-850nm, some extend to 350-900nm, or even 200-1500nm, it is very difficult to correct chromatic aberration in such a wide spectral range.

[0126] The optical signal collecting lens 250 introduces a spherical reflective surface 264, and optimizes the center of its curvature to a non-spherical deflecting surface 262 to correct spherical aberration, and the deflecting surface 262 acts as a stop, so that the system has a larger uniform image field. Since the structure of the flow-type liquid flow cavity is highly light-transmissive and its material is generally made of optical glass, part of the surface of the optical lens can be made into an integral structure with the liquid flow cavity, such as the first transmission part 260, which can make the optical material composed of the refractive surface 263 and the reflective surface 264 glued together by optical gel or molecular bonding. If the fluid channel 261 is made of the same material, then the refractive surface 263 is not needed, and the reflective surface 264 and the deflecting surface 262 are directly optically processed. Specifically, CNC grinding or other processing methods can be adopted to optimize the refractive surface 263 and the deflecting surface 262 to a reasonable surface shape. By processing the refractive surface 263 and the deflecting surface 262 of the optical lens separately from the fluid channel 261, the processing yield can be guaranteed. The reflective surface 264 may be a spherical surface, a parabolic surface or other aspherical surfaces. Considering the difficulty of processing and the sensitivity of manufacturing errors, a spherical surface is used in this embodiment, and the deflecting surface 262 is used to correct the spherical aberration introduced by the reflective surface 264. The residual aberration is the change of spherical aberration with wavelength or some high-order astigmatism and the oblique light spherical aberration introduced by off-axis light. Although the reflective surface 264 and the liquid flow cavity of the same material can reduce the chromatic aberration to a relatively low level, since the light first emerges from the aqueous solution to the glass and then from the glass to the air, the change of the refractive index of the material will also produce certain chromatic aberration or secondary spectrum in the wide spectrum domain. The optical signal collection lens 250 is provided with a second transmission part 270, so that the lens of the second transmission part 270 uses flint glass and crown glass with high and low refractive indices to work together to correct the chromatic aberration or secondary spectrum, and the number and thickness of the flint glass and crown glass are not limited, and it can be a double cemented lens or a triple cemented lens.

[0127] The second transmission part 270 is a triple-cemented lens, which is composed of two pieces of high-refractive-index flint glass and one piece of low-refractive-index biconvex crown glass, and is configured to correct the remaining chromatic aberration and coma astigmatism, etc. and focus the light beam. The flint glass materials of the first sub-lens 271, the second sub-lens 272 and the third sub-lens 273 can be the same material or different materials, and there is a certain degree of freedom in selection. The central low-refractive-index crown glass is generally K9 or BK7 glass, which has the characteristics of low cost and stable performance.

[0128] There is air between the first transmission part 260 and the second transmission part 270 to serve as a spacer. Since the optical system 1200 composed of the reflection and deflection surface 262 is in an infinite conjugate form, there is no restriction on the air spacing distance between the two parts. Corresponding optical elements or other devices can be inserted in the middle, which increases the freedom in optical and structural design.

[0129] It is worth noting that the effects that can be achieved by the optical signal collection lens 250 include: numerical aperture reaches 1.21, field of view reaches 500um, workpiece distance is greater than 2mm, RMS radius of the focusing light plate is less than 50um, wavelength covers 400nm~900nm, and magnification is 9 times. By limiting various parameters, an optical signal collection lens 250 with high numerical aperture, long working distance and large field of view is obtained.

[0130] It should be noted that the specific setting parameters of the optical signal collection lens 250 in this embodiment can refer to the following Table 1 (unit: mm):

[0131]

[0132] Table 1

[0133] It is worth noting that the radius, interval and refractive index parameters in Table 1 are mostly range values, and the preferred values ​​are usually middle values ​​close to the range values, but the specific selected values ​​in the design process can be kept within the allowable range.

[0134] This embodiment provides a control method for a flow cytometer, which is applied to control the flow cytometer. The control method specifically includes:

[0135] Step S100: controlling the negative pressure tank 100 to form a negative pressure, and loading the sample 2000 into the flow chamber 300 through the sampling needle 400;

[0136] Step S200 : controlling the sheath liquid pump 200 to deliver sheath liquid to the flow chamber 300 , so that the sheath liquid wraps the sample 2000 .

[0137] Specifically, step S100 includes:

[0138] Step S110: controlling the negative pressure formed in the negative pressure tank 100 to be constant;

[0139] Step S120 : controlling the sampling needle 400 to draw the sample 2000 and load it into the flow chamber 300 .

[0140] It should be noted that the above steps S100 and S200 can be performed in the order of S100 first and S200 later, or in the order of S100 later and S200 first, or both can be performed simultaneously. The specific operation order can be determined according to the actual collection situation.

[0141] In addition, in order to further ensure the accuracy and stability of the overall operation, the pressure of the negative pressure tank 100 can be maintained constant during the operation, and the flow rate of the sample 2000 can be adjusted by adjusting the sheath liquid flow rate. The advantage of this control method is that it can ensure the stability of the core flow, and the stability of the core flow is reflected in the width, position and flow rate of the core flow. Under normal loading conditions, the position of the core flow rarely shakes and changes, unless the flow rate of the sample 2000 or the sheath liquid flow rate fluctuates greatly. One of the factors that mainly determines the width and flow rate of the core flow is the flow rate of the sample 2000. By keeping the pressure of the negative pressure tank 100 constant as much as possible, and making the sheath liquid pump 200 control the stability of the flow rate of the sample 2000, and the flow rate of the sample 2000 actually depends on the pressure in the flow chamber 300, it can also be understood that the stability of the flow rate of the sample 2000 controlled by the sheath liquid pump 200 is to first control the stability of the pressure in the flow chamber 300, and the stability of the pressure of the flow chamber 300 and the pressure of the negative pressure tank 100 naturally also brings about the stability of the liquid flow in the flow pool 310.

[0142] Wherein, step S110 includes:

[0143] Step S111: obtaining the real-time pressure in the negative pressure tank 100;

[0144] Step S112: Control the waste liquid pump 500 to perform PID regulation on the pressure in the negative pressure tank 100. Step S200 includes:

[0145] Step S210: Control the sheath liquid pump 200 to perform PID control on the flow rate of the sample 2000 entering the sampling needle 400.

[0146] Specifically, the first pressure sensor 140 can be used for feedback and closed-loop PID control of the waste liquid pump 500 (PID control is a control system based on three control methods: proportional, integral and differential. The error between the real-time data of the controlled object and the given value can be adjusted, and the three control methods of proportional, integral and differential can be used to eliminate the error and achieve the control target) so that the negative pressure tank 100 maintains a constant negative pressure; similarly, the flow meter 70 feedback and the sheath liquid pump 200 are used for closed-loop PID control to control and adjust the loading speed. Moreover, the two PID control methods are relatively independent, and the target value and feedback value belong to the same attribute value, without the need for indirect conversion, and thus have the characteristics of simpler control and faster real-time feedback. Moreover, the structure that requires dual light sources and dual channels in the prior art can be avoided, so that the structure is also simpler.

[0147] It is worth noting that the existing technology measures the laser delay or sample flow rate through a dual light source device and a dual scattering channel, and then derives and converts the pressure difference reference, and indirectly achieves the stability of the total volume flow rate and the sample flow rate by controlling the pressure difference between the two ends of the flow channel of the flow chamber. The stability of the total volume flow rate controlled by this adjustment method is not equivalent to the stability of the sample flow rate, because the total volume passing through the flow channel is composed of sheath fluid and sample. Although the total volume flow rate is kept stable, the proportional relationship between the sample flow rate and the sheath fluid flow rate will still change, thereby affecting the accuracy of the test results. In addition, the target value and feedback value of this adjustment method do not belong to the same attribute value, and need to be indirectly converted, which makes the control more complicated, the real-time feedback slower, and the structure more complicated.

[0148] Please refer to Figure 8 According to the above control method, the target pressure value of the negative pressure tank 100 is P1, and P1 is controlled to be a value between -60Kpa and -10Kpa. PID is used to adjust the pressure in the negative pressure tank 100, that is, the pressure in the negative pressure tank 100 is controlled to be dynamically constant and maintained at P1. The lower the target pressure value of the negative pressure tank 100, the more time it takes for the negative pressure tank 100 to establish negative pressure, and the longer the corresponding flow cytometer preparation time and the lower the efficiency; reducing the volume of the negative pressure tank 100 can shorten the time to establish negative pressure, but reducing the volume of the negative pressure tank 100 will increase the pressure fluctuation in the negative pressure tank 100, thereby increasing the fluctuation of the sample flow and the core flow; increasing the displacement of the waste liquid pump 500 can also shorten the time to establish negative pressure, but the control accuracy of the large-displacement waste liquid pump 500 is necessarily lower. The target pressure value for PID control of the pressure in the negative pressure tank 100 is set within this range, which comprehensively meets the factors such as the preparation time for establishing negative pressure before loading, the influence of adjustment on pressure fluctuation, and the adjustment accuracy control of the waste liquid pump 500.

[0149] The flow of the sample 2000 entering the sampling needle 400 is PID controlled by the sheath liquid pump 200, and the flow value is set to the target value. During the detection of the sample 2000, the flow value is set to Q1, wherein 5ul / min≤Q1≤200ul / min. The interval setting of the flow of the sample 2000 is mainly determined by the speed and accuracy of the sample loading detection. When the flow of the sample 2000 is too low, the collection time will be prolonged. When the flow of the sample 2000 is too high, the core flow size will be too large, thereby reducing the detection accuracy.

[0150] Specifically, the flow rate of the sheath fluid flowing to the flow chamber 300 can be Q2, and the flow resistance of the sample 2000 flowing through the sampling channel section can be R1, and the flow resistance of the fluid channel in the flow cell can be R3, and can be combined with the formula Q1=(-P1-Q2*R3) / (R1+R3), wherein R1, R3, and P1 are all constant values, that is, the purpose of adjusting Q1 can be achieved by adjusting Q2.

[0151] During the sample loading test, by maintaining the pressure P1 of the negative pressure tank 100 constant and controlling the flow rate Q1 of the sample 2000 by the sheath liquid pump 200 PID to maintain stability, the stability of the core flow size, position and liquid flow can be guaranteed, and the accuracy of the test results can be ensured. In addition, by adjusting the sheath liquid flow rate Q2 by the sheath liquid pump 200, the sample flow rate Q1 can be changed to achieve different sample loading test speed requirements.

[0152] Step 400: Control the light source to illuminate the sample 2000 and generate a light beam, and enable the optical signal collection lens 250 to transmit and collect the light beam.

[0153] Of course, in order to improve the cleanliness of the sampling needle 400 and avoid the negative impact on the sampling process due to the low cleanliness of the sampling needle 400, the control method may also include step S500: controlling the sheath fluid bottle 700 to deliver sheath fluid to the cleaning channel 611 to clean the outer wall of the sampling needle 400.

[0154] In addition, in order to achieve stable sample loading operation, the control method can start the waste liquid pump 500 and the sheath liquid pump 200, open the first valve 111 and the fifth valve 115 at the same time, and provide negative pressure through the waste liquid pump 500, so that the needle can suck the sample into the flow chamber 300. At the same time, the sheath liquid pump 200 transports the sheath liquid to the flow chamber 300, so that the sheath liquid wraps the sample 2000 to form a laminar flow for optical detection.

[0155] Afterwards, the waste liquid enters into the negative pressure tank body 120 of the negative pressure tank 100 through the first valve 111 , and then enters into the waste liquid bottle 50 through the waste liquid pump 500 .

[0156] It can be seen that in this embodiment, the negative pressure source is formed by the cooperation of the negative pressure tank 100 and the waste liquid pump 500, so that a continuous sampling measurement process can be realized. At the same time, it replaces the injection pump and the plunger pump used in the prior art to carry out the sampling process, simplifies the pipeline connection, and reduces the liquid circuit volume.

[0157] In addition, it should be noted that the negative pressure tank 100 and the damping device 130 can effectively reduce the pulsation generated by the waste liquid pump 500, improve the stability of the liquid flow, and improve the detection accuracy. At the same time, the pressure buffered by the negative pressure tank 100 can reduce the startup time for starting sample loading and improve the detection efficiency of the flow cytometer.

[0158] In order to facilitate cleaning of the outer wall of the sampling needle 400, the control method provided in this embodiment further includes:

[0159] The swab pump 620 and the sheath fluid pump 200 are controlled to work so that the sheath fluid in the sheath fluid bottle 700 flows through the cleaning channel 611 and the outer wall of the sampling needle 400. At the same time, the sampling needle 400 and the swab 610 are controlled to move relative to each other to clean the outer wall of the sampling needle 400.

[0160] Specifically, by starting the swab pump 620 and the sheath fluid pump 200, and opening the fourth valve 114 and the sixth valve 60 at the same time, the sheath fluid from the sheath fluid bottle 700 passes through the sheath fluid pump 200, the depulsation device 10, the bypass of the first filter 900 and the fourth valve 114 in sequence and then enters the main channel of the swab 610. At the same time, the sampling needle 400 and the cleaning channel 611 slide relative to each other to achieve cleaning of the outer wall of the sampling needle 400. The waste liquid generated by the cleaning passes through the sixth valve 60 and the swab pump 620 and flows into the waste liquid bottle 50.

[0161] It should be noted that after each sample 2000 loading process is completed, the outer wall of the sampling needle 400 needs to be cleaned. Since the sheath fluid will flow through the first filter 900, each time the outer wall of the sampling needle 400 is cleaned, the bubble removal process of the first filter 900 is also completed.

[0162] In order to facilitate the cleaning of the inner wall of the sampling needle 400, the control method further includes:

[0163] The swab pump 620 and the sheath fluid pump 200 are controlled to operate so as to drive the sheath fluid in the sheath fluid bottle 700 to flow through the sheath fluid pump 200 , the flow chamber 300 , the internal sampling channel of the sampling needle 400 , the cleaning channel 611 and the swab pump 620 in sequence.

[0164] Specifically, by starting the swab pump 620 and the sheath fluid pump 200, opening the fifth valve 115 and the sixth valve 60, the sheath fluid from the sheath fluid bottle 700 passes through the sheath fluid pump 200, the depulsation device 10, the main channel of the first filter 900 (not the bypass of the first filter 900 mentioned above), the degasser 800 and the fifth valve 115, and then enters the flow chamber 300, and then enters the interior of the sampling needle 400 to clean the inner wall of the sampling needle 400. After that, the waste liquid passes through the first sub-channel 6111, the sixth valve 60 and the swab pump 620 and enters the waste liquid bottle 50.

[0165] In addition, in order to facilitate cleaning of the flow chamber 300 and the sampling channel of the sampling needle 400 by the cleaning liquid, the control method further includes:

[0166] First, the cleaning liquid pump 20 is controlled to work to drive the cleaning liquid in the cleaning liquid bottle 30 to enter the internal sampling channel of the flow chamber 300 and the sampling needle 400. Specifically, the cleaning liquid pump 20 can be started first to transport the cleaning liquid in the cleaning liquid bottle 30 to the sampling channel inside the flow chamber 300 and the sampling needle 400.

[0167] After the cleaning liquid is delivered, the cleaning liquid can be allowed to stay in the flow chamber 300 and the internal sampling channel of the sampling needle 400 and soak for a certain period of time (the time can be set according to the specific situation) to improve the cleaning effect.

[0168] Then, the waste liquid discharge process is carried out, and one of the waste liquid discharge methods is: by controlling the operation of the waste liquid pump 500, at the same time, opening the first valve 111, so that negative pressure is formed in the negative pressure tank 100, so as to discharge the waste liquid in the flow chamber 300 and the internal sampling channel of the sampling needle 400. The waste liquid passes through the first valve 111, the negative pressure tank 100 and the waste liquid pump 500 in turn and enters the waste liquid bottle 50.

[0169] Another way to discharge the waste liquid is to control the swab pump 620 to open the sixth valve 60 to discharge the waste liquid in the flow chamber 300 and the internal sampling channel of the sampling needle 400. The waste liquid passes through the first subchannel 6111, the sixth valve 60 and the swab pump 620 in turn and enters the waste liquid bottle 50.

[0170] In addition, in order to realize the debubbling process of the flow chamber 300, the control method further includes:

[0171] First, the flow chamber 300 is emptied, and the swab pump 620 is controlled to work so that external gas enters the negative pressure tank 100 , the flow chamber 300 and the swab pump 620 in sequence to empty the flow chamber 300 .

[0172] Specifically, the swab pump 620 is started, the first valve 111, the second valve 112 and the third valve 113 are opened, and the outside air enters the negative pressure tank 100 through the air intake filter 90 and the third valve 113, and then enters the flow chamber 300 through the first valve 111. The liquid in the flow chamber 300 then enters the waste liquid bottle 50 after passing through the second valve 112 and the swab pump 620, thereby completing the process of emptying the flow chamber 300.

[0173] It should be noted that when the flow chamber 300 needs to be unblocked, the above-mentioned liquid path control method for emptying the flow chamber 300 can also be used.

[0174] Afterwards, the process of filling the flow chamber 300 is performed. Specifically, the control method further includes:

[0175] The waste liquid pump 500 and / or the sheath liquid pump 200 are controlled to work so that the sheath liquid passes through the flow chamber 300 , the negative pressure tank 100 and the waste liquid pump 500 in sequence to fill the flow chamber 300 .

[0176] Specifically, by opening the fifth valve 115 and the first valve 111, the waste liquid pump 500 and the sheath liquid pump 200 can be started, or the waste liquid pump 500 or the sheath liquid pump 200 can be started, and the sheath liquid in the sheath liquid bottle 700 passes through the sheath liquid pump 200, the first filter 900, the degasser 800 and the fifth valve 115 in sequence and enters the flow chamber 300, and then passes through the first valve 111, the interior of the negative pressure tank 100 and the waste liquid pump 500 and enters the waste liquid bottle 50, thereby filling the flow chamber 300 and completing the process of removing bubbles from the flow chamber 300.

[0177] It should be noted that when the flow chamber 300 needs to be unblocked, the above-mentioned liquid path control method for filling the flow chamber 300 can also be used.

[0178] That is, in this embodiment, the swab pump 620 is controlled to work so that the external gas enters the negative pressure tank 100, the flow chamber 300 and the swab pump 620 in sequence, and the step of emptying the flow chamber 300 is the first step; the waste liquid pump 500 and / or the sheath liquid pump 200 are controlled to work so that the sheath liquid passes through the flow chamber 300, the negative pressure tank 100 and the waste liquid pump 500 in sequence, and the step of filling the flow chamber 300 is the second step. The control method also includes: the first step is located before the second step, that is, the first step is performed first, and then the second step is completed, so as to realize the process of removing bubbles from the flow chamber 300.

[0179] In order to eliminate the blockage of the sampling channel, the control method further includes:

[0180] The waste liquid pump 500 is controlled to work, so that negative pressure is formed in the negative pressure tank 100, so that the sampling needle 400 extracts the blocking liquid, cleaning liquid or air, and flows through the flow chamber 300, the negative pressure tank 100, the waste liquid pump 500 and the waste liquid bottle 50 in sequence.

[0181] Specifically, the waste liquid pump 500 is started and the first valve 111 is opened at the same time, so that the sampling needle 400 extracts the draining liquid, cleaning liquid or air, and the liquid flows through the flow chamber 300, the first valve 111, the interior of the negative pressure tank 100 and the waste liquid pump 500 in sequence, and then enters the waste liquid bottle 50.

[0182] In addition, it should be noted that the above control methods can be used in combination to complete complex actions such as powering on, powering off, and deep cleaning of the flow cytometer.

[0183] For example, during the startup process of the flow cytometer, the first filter 900 can be debubbled, and at the same time, the flow chamber 300 can also be debubbled, so as to facilitate the debubbling process of the liquid path and improve the detection stability.

[0184] During the shutdown process of the flow cytometer, the above-mentioned cleaning process of the flow chamber 300 can be performed. In addition, deep cleaning can also be achieved.

[0185] In summary, the flow cytometer, optical signal collection lens, and sample analysis device provided in this embodiment have at least the following advantages:

[0186] 1. Negative pressure is provided by the waste liquid pump 500, and the sheath liquid supply is controlled by the sheath liquid pump 200, so as to adjust the loading speed of the sample 2000. The flow rate of the sample 2000 is detected by the flow meter 70. While realizing absolute volume counting, continuous loading and measurement can be performed. Compared with the existing injection pump and plunger pump loading methods, the pipeline connection is simple and the liquid path volume is greatly reduced.

[0187] 2. The negative pressure tank 100 and the waste liquid pump 500 cooperate to form a negative pressure source. The negative pressure tank 100 can realize the function of rapid pressure relief. At the same time, the negative pressure tank 100 has a damping device 130, which is equivalent to a pressure stabilizing device or a buffer, etc. The damping device 130 facilitates the precise control of the pressure in the negative pressure tank 100 and effectively reduces pressure fluctuations. In addition, the negative pressure tank 100 can realize real-time emptying of the liquid in the negative pressure tank 100 without storing waste liquid, so there is no need to set a liquid level sensor. At the same time, the waste liquid pump 500 can use a diaphragm pump with lower precision and lower cost, and can also accurately control the pressure in the negative pressure tank 100 to achieve an effect of smaller output pressure fluctuations.

[0188] 3. The de-pulsation device 10 can effectively remove the liquid flow pulsation generated by the sheath liquid pump 200, thereby improving the stability of the liquid flow and the detection accuracy.

[0189] 4. The flow chamber 300 and the sampling channel can be effectively cleaned by the cleaning liquid pump 20, thereby improving the consistency of the test results, reducing the carryover contamination rate, and making the flow cytometer more functional and more automated.

[0190] 5. The negative pressure tank 100 can effectively reduce the pulsation generated by the waste liquid pump 500, improve the stability of the liquid flow, and improve the detection accuracy. The pressure buffered by the negative pressure tank 100 can reduce the start-up time of the collection and improve the detection efficiency of the flow cytometer.

[0191] 6. The degasser 800 is used to remove the air content in the sheath liquid, which solves the influence of bubbles on the liquid flow, improves the stability of the liquid flow, improves the detection accuracy, and expands the adaptability of the flow cytometer to different usage environments.

[0192] The control method of the flow cytometer mentions the method of first aspirating the sample and then loading the sample adopted in the prior art, realizing a continuous loading process. At the same time, the operation and cleaning process are simple, which solves the problems existing in the prior art that the pipeline connection, sample aspiration, loading and cleaning are relatively complicated, resulting in poor consistency of detection results and high carryover contamination rate.

[0193] The above is only a specific embodiment of the present disclosure, but the protection scope of the present disclosure is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present disclosure should be included in the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be based on the protection scope of the claims.

[0194] Industrial Applicability

[0195] Compared with the prior art, the flow cytometer improves the sample loading method, simplifies the operation procedure, and realizes continuous sample loading.

Claims

1. A flow cytometer, characterized in that: include: A fluid system (1100) and an optical system (1200); The liquid circuit system (1100) comprises a sampling needle (400), a sheath fluid bottle (700) and a flow chamber (300); the sampling needle (400) and the sheath fluid bottle (700) are both in communication with the flow chamber (300) and are respectively configured to transport a sample (2000) and sheath fluid to the flow chamber (300); The optical system (1200) comprises a light source (240) and a light signal collecting lens (250), wherein the light signal collecting lens (250) comprises a fluid channel (261), wherein the fluid channel (261) is connected to the flow chamber (300), wherein the light source (240) is configured to irradiate a sample (2000) flowing from the flow chamber (300) into the fluid channel (261), and wherein the light signal collecting lens is configured to collect and transmit a light beam generated by the sample (2000) after being irradiated; The liquid circuit system (1100) further comprises a negative pressure tank (100) and a waste liquid pump (500), wherein the waste liquid pump (500) is connected to the flow chamber (300), and the negative pressure tank (100) is arranged between the waste liquid pump (500) and the flow chamber (300). The waste liquid pump (500) operates to form a negative pressure in the negative pressure tank (100), so that the sample (2000) is loaded from the sampling needle (400) into the flow chamber (300) and enters the fluid channel (261).

2. The flow cytometer according to claim 1, characterized in that The waste liquid pump (500) is controlled to maintain the pressure in the negative pressure tank (100) dynamically constant at a target pressure value.

3. The flow cytometer according to claim 2, characterized in that The negative pressure tank (100) comprises a first pressure sensor (140), which is configured to obtain a real-time pressure value in the negative pressure tank (100), and use the real-time pressure value as a feedback value and the target pressure value as a target value to perform PID control on the pressure in the negative pressure tank (100) through the waste liquid pump (500).

4. The flow cytometer according to claim 2, characterized in that The target pressure value is P1, wherein -60Kpa≤P1≤-10Kpa.

5. The flow cytometer according to any one of claims 1 to 4, characterized in that: The liquid path system (1100) also includes a sheath liquid pump (200), which is connected between the sheath liquid bottle (700) and the flow chamber (300). The sheath liquid supply is adjusted by controlling the sheath liquid pump (200) to adjust the flow rate of the sample (2000) flowing to the flow chamber (300).

6. The flow cytometer according to claim 5, characterized in that The liquid circuit system (1100) further comprises a flow meter (70), wherein the flow meter (70) is arranged on a pipeline between the sampling needle (400) and the flow chamber (300), and the flow meter (70) is configured to obtain a real-time flow value of the sample (2000) flowing to the flow chamber (300), and to perform PID control on the flow of the sample (2000) entering the sampling needle (400) by using the real-time flow value as a feedback value and a set flow value as a target value through the sheath liquid pump (200).

7. The flow cytometer according to claim 6, characterized in that During the sample detection process, the set flow value is Q1, wherein 5ul / min≤Q1≤200ul / min.

8. The flow cytometer according to any one of claims 1 to 7, characterized in that: The negative pressure tank (100) comprises a negative pressure tank body (120) and a damping device (130), wherein the damping device (130) is arranged on the negative pressure tank body (120), and the damping device (130) is connected to the negative pressure tank body (120); And / or, the negative pressure tank (100) comprises a negative pressure tank body (120), the negative pressure tank body (120) is provided with a first flow channel (122) and a pressure relief port (121), one end of the first flow channel (122) is connected to the pressure relief port (121), the other end of the first flow channel (122) is configured to be connected to the inside of the negative pressure tank body (120), and the pressure relief port (121) is configured to achieve rapid pressure relief of the negative pressure tank (100); And / or, the negative pressure tank (100) comprises a negative pressure tank body (120) and a control valve (110), the negative pressure tank body (120) is provided with a first flow path channel (122) and a pressure relief port (121), and the control valve (110) is arranged in the first flow path channel (122) and is configured to realize the connection and disconnection between the negative pressure tank body (120) and the pressure relief port (121).

9. The flow cytometer according to any one of claims 1 to 8, characterized in that: The liquid circuit system (1100) further comprises a swab (610), wherein the swab (610) is provided with a cleaning channel (611), and the sampling needle (400) is slidably matched with the cleaning channel (611); Preferably, the cleaning channel (611) is slidable relative to the sampling needle (400).

10. The flow cytometer according to claim 9, characterized in that: The liquid path system (1100) further comprises a swab pump (620), one end of the cleaning channel (611) is in communication with the swab pump (620), and the other end of the cleaning channel (611) is in communication with the sheath fluid bottle (700); And / or, the liquid path system (1100) further includes a cleaning liquid pump (20) and a cleaning liquid bottle (30), one end of the cleaning liquid pump (20) is connected to the cleaning liquid bottle (30), and the other end of the cleaning liquid pump (20) is connected to the flow chamber (300).

11. The flow cytometer according to any one of claims 1 to 10, characterized in that: The liquid circuit system (1100) further comprises a waste liquid bottle (50), wherein the waste liquid bottle (50) is in communication with the waste liquid pump (500) and is configured to receive waste liquid output from the negative pressure tank (100); And / or, the liquid circuit system (1100) further comprises a sheath liquid pump (200), a degasser (800) and a first pipeline (190), wherein the sheath liquid bottle (700), the sheath liquid pump (200), the degasser (800) and the flow chamber (300) are sequentially arranged in the first pipeline (190); And / or, the fluid path system (1100) comprises a sheath liquid pump (200) and a depulsation device (10), one end of the depulsation device (10) is connected to the sheath liquid pump (200), and the other end of the depulsation device (10) is connected to the flow chamber (300).

12. The flow cytometer according to any one of claims 1 to 11, characterized in that: The optical signal collecting lens (250) comprises a first transmission part (260), a second transmission part (270) and a collecting part (280) which are sequentially arranged at intervals; The first transmission part (260) is provided with the fluid channel (261), and the fluid channel (261) is configured to transport the sample (2000). The sample (2000) generates a lateral light beam (246) after being irradiated by the laser. A deflection surface (262) is formed on one side of the first transmission part (260) close to the second transmission part (270), and the deflection surface (262) is configured to correct the spherical aberration of the lateral light beam (246).

13. The flow cytometer according to claim 12, characterized in that: The deflecting surface (262) is wave-shaped and concave; And / or, the first transmission part (260) further includes a reflection surface (264), the reflection surface (264) is a curved surface and is located on a side of the fluid channel (261) away from the second transmission part (270); And / or, the first transmission portion (260) further includes a reflective surface (264) and a refractive surface (263), wherein the refractive surface (263) is a plane and is located between the reflective surface (264) and the fluid channel (261); And / or, the first transmission part (260) further comprises a reflection surface (264), the reflection surface (264) is coated with a reflection film (265), and the reflection film (265) is a dielectric film or a metal film.

14. The flow cytometer according to claim 13, characterized in that The cross section of the fluid channel (261) is rectangular, and the lateral edge of the reflecting surface (264) is located on the extension line of the diagonal line of the same rectangular cross section as the fluid channel (261); Preferably, the side length of the rectangular cross section of the fluid channel (261) is between 100um and 500um.

15. An optical signal collecting lens, characterized in that: include: A first transmission part (260), a second transmission part (270) and a collection part (280) are arranged in sequence and at intervals; The first transmission part (260) is provided with a fluid channel (261), and the fluid channel (261) is configured to transport a sample (2000). The sample (2000) generates a lateral light beam (246) after receiving irradiation of a laser. A deflection surface (262) is formed on a side of the first transmission part (260) close to the second transmission part (270), and the deflection surface (262) is configured to correct spherical aberration of the lateral light beam (246).

16. The optical signal collecting lens according to claim 15, characterized in that: The deflecting surface (262) is in a wave-shaped concave shape.

17. The optical signal collecting lens according to claim 15, characterized in that: The first transmission part (260) further comprises a reflection surface (264), wherein the reflection surface (264) is a curved surface and is located on a side of the fluid channel (261) away from the second transmission part (270).

18. The optical signal collecting lens according to claim 17, characterized in that: The first transmission portion (260) further comprises a refractive surface (263), wherein the refractive surface (263) is a plane and is located between the reflective surface (264) and the fluid channel (261); And / or, the reflective surface (264) is coated with a reflective film (265), and the reflective film (265) is a dielectric film or a metal film.

19. The optical signal collecting lens according to any one of claims 15 to 18, characterized in that: The second transmission part (270) is a triplet lens, and comprises a first sub-lens (271), a second sub-lens (272), and a third sub-lens (273) which are sequentially arranged along a direction from the collecting part (280) to the first transmission part (260).

20. The optical signal collecting lens according to claim 19, characterized in that: The first sub-lens (271) is arc-shaped, and the middle position of the first sub-lens (271) is closer to the collecting portion (280) relative to both ends; And / or, the third sub-lens (273) is arc-shaped, and the middle position of the third sub-lens (273) is closer to the first transmission part (260) relative to both ends.

21. A sample analysis device, characterized in that: It comprises the flow cytometer described in any one of claims 1 to 14 or the optical signal collecting lens described in any one of claims 15 to 20.

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