Flow cytometer, optical signal acquisition lens and sample analysis device

Abstract

A flow cytometer, an optical signal acquisition lens and a sample analysis device. The flow cytometer comprises a liquid path system (1100) and an optical system (1200). The liquid path system (1100) comprises a sampling needle (400), a sheath liquid bottle (700) and a flow chamber (300), the sampling needle (400) and the sheath liquid bottle (700) being both communicated with the flow chamber (300) and being respectively configured to deliver a sample (2000) and a sheath liquid to the flow chamber (300). The optical system (1200) comprises a light source (240) and an optical signal acquisition lens (250), the optical signal acquisition lens (250) comprising a fluid channel (261), and the fluid channel (261) being communicated with the flow chamber (300). The liquid path system (1100) further comprises a negative pressure tank (100) and a waste liquid pump (500), the waste liquid pump (500) being communicated with the flow chamber (300), and the negative pressure tank (100) being arranged between the waste liquid pump (500) and the flow chamber (300). The waste liquid pump (500) is operated to form a negative pressure in the negative pressure tank (100), so that a sample (2000) is loaded from the sampling needle (400) to the flow chamber (300) and enters the fluid channel (261). By means of the sample loading mode, the flow cytometer simplifies the operation procedure and achieves continuous sample loading.

Description

A flow cytometer, an optical signal collection lens and a sample analysis device

[0001] Cross-reference to Related Applications

[0002] The present disclosure claims priority to the Chinese patent application No. 202410879963.7, filed on July 2, 2024, entitled "A optical signal collection lens and a sample analysis device", the entire content of which is incorporated herein by reference. TECHNICAL FIELD

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

[0004] The flow cytometer is generally configured to analyze and count cells or particles in a sample. The flow cytometer mainly consists of a liquid system, an optical system, and a data analysis system. The liquid system, as one of the important components of the flow cytometer, transports the sample containing cells or particles labeled by fluorescent dye and sheath fluid to the flow chamber, focuses the cells or particles by fluid dynamics, and makes the cells or particles pass through the detection area of the flow chamber one by one. Stable sample flow is the guarantee for subsequent signal detection and is one of the core technologies of the entire instrument.

[0005] The mainstream flow cytometers on the current market mostly have absolute counting function. Absolute volume counting is to measure the volume of the sample, then divide the number of measured cells by the volume of the sample, so as to obtain the concentration information of the sample and achieve the purpose of absolute counting. The absolute volume counting scheme is mainly realized by a quantitative pump that can quantitatively transport the sample volume, such as a syringe pump, a plunger pump, and a peristaltic pump. The pipeline connection, sample suction, sample loading, and cleaning are relatively complex, which may cause poor consistency of detection results, high contamination rate, and other problems. The syringe pump and the plunger pump cannot continuously load the sample, and need to first suck the sample and then load the sample, which is a complex operation procedure. SUMMARY

[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 continuously load the sample.

[0007] Embodiments of the present disclosure can be implemented as follows:

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

[0009] a liquid system and an optical system;

[0010] The liquid path system comprises a sampling needle, a sheath liquid bottle and a flow chamber, the sampling needle and the sheath liquid bottle are communicated with the flow chamber and are respectively configured to transport sample and sheath liquid to the flow chamber;

[0011] The optical system comprises a light source and a light signal collection lens, the light signal collection lens comprises a fluid channel, the fluid channel is communicated with the flow chamber, the light source is configured to irradiate the sample flowing into the fluid channel from the flow chamber, and the light signal collection lens is configured to collect and transmit the light beam generated by the sample after being irradiated;

[0012] 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 the waste liquid pump is operated to form negative pressure in the negative pressure tank, so that the sample is loaded from the sampling needle to the flow chamber and into 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 comprises a first pressure sensor configured to obtain a real-time pressure value in the negative pressure tank, the real-time pressure value is taken as a feedback value, and a target pressure value is taken as a target value, and the pressure in the negative pressure tank is PID controlled by the waste liquid pump.

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

[0016] Optionally, the liquid path system further comprises a sheath liquid pump, the sheath liquid pump is communicated between the sheath liquid bottle and the flow chamber, and the sheath liquid pump is controlled to adjust the sheath liquid supply amount to adjust the flow rate of the sample flowing to the flow chamber.

[0017] Optionally, the liquid path system further comprises a flow meter, the flow meter is arranged on a pipeline between the sampling needle and the flow chamber, the flow meter is configured to obtain a real-time flow value of the sample flowing to the flow chamber, the real-time flow value is taken as a feedback value, a set flow value is taken as a target value, and the flow of the sample entering the sampling needle is PID controlled by 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, the damping device is arranged in the negative pressure tank body, and the damping device and the negative pressure tank body are communicated;

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

[0021] And / or, the negative pressure tank comprises 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, and the control valve is arranged in the first flow path channel and is configured to realize the on-off of the negative pressure tank body and the pressure relief port.

[0022] Optionally, the liquid path system further comprises a swab, the swab is provided with a cleaning channel, and the sampling needle is in sliding fit with the cleaning channel.

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

[0024] Optionally, the liquid path system further comprises a swab pump, one end of the cleaning channel is communicated with the swab pump, and the other end of the cleaning channel is communicated with the sheath liquid bottle.

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

[0026] Optionally, the liquid path system further comprises a waste liquid bottle, the waste liquid bottle is communicated with the waste liquid pump and is configured to receive the waste liquid output from the negative pressure tank.

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

[0028] And / or, the liquid path system comprises a sheath liquid pump and a de-pulsation device, one end of the de-pulsation device is communicated with the sheath liquid pump, and the other end of the de-pulsation device is communicated with the flow chamber.

[0029] Optionally, the light signal collection lens comprises a first transmission part, a second transmission part and a collection part arranged in sequence and at intervals.

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

[0031] Optionally, the baffle surface is in a wave-shaped concave type.

[0032] And / or, the first transmission part further comprises a reflecting surface, the reflecting surface is arc-shaped and located on the side of the fluid channel away from the second transmission part.

[0033] And / or, the first transmission part further comprises a reflecting surface and a refracting surface, the refracting surface is planar and located between the reflecting surface and the fluid channel.

[0034] And / or, the first transmission part further comprises a reflecting surface, the reflecting surface is coated with a reflecting film, the reflecting 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 reflecting surface is located on the extension line of the diagonal of the same rectangular cross section of 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] Embodiments of the present disclosure also provide a light signal collection lens, comprising:

[0038] A first transmission part, a second transmission part and a collection part are sequentially and spaced apart.

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

[0040] Optionally, the baffling surface is in a wavy concave shape.

[0041] Optionally, the first transmission part further comprises a reflecting surface, the reflecting surface is arc-shaped and located on the side of the fluid channel away from the second transmission part.

[0042] Optionally, the first transmission part further comprises a refracting surface, the refracting surface is planar and located between the reflecting surface and the fluid channel.

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

[0044] Optionally, the second transmission part is a three-glued lens and comprises a first sub-lens, a second sub-lens and a third sub-lens sequentially arranged in the direction from the collection part to the first transmission part.

[0045] Optionally, the first sub-lens is in an arc shape, and the middle position of the first sub-lens is closer to the collection part relative 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 part than two ends.

[0047] Embodiments of the present disclosure also provide a sample analysis device, which comprises the flow cytometer or the optical signal analysis lens.

[0048] The flow cytometer of the embodiments of the present disclosure has the following beneficial effects, for example:

[0049] 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 flow chamber, the sampling needle and the sheath liquid bottle are both in communication with the flow chamber and are respectively configured to deliver a sample and a sheath liquid to the flow chamber; the optical system comprises a light source and an optical signal collection lens, the optical signal collection lens comprises a fluid channel, the fluid channel is in communication with the flow chamber, the light source is configured to irradiate the sample flowing from the flow chamber into the fluid channel, and the optical signal collection lens is configured to collect and transmit a light beam generated by the sample after being irradiated; wherein the liquid path system further comprises a negative pressure tank and a waste liquid pump, the waste liquid pump is in communication with the flow chamber, and the negative pressure tank is arranged between the waste liquid pump and the flow chamber; the waste liquid pump is operated to form a negative pressure in the negative pressure tank, so that the sample is loaded from the sampling needle to the flow chamber and into the fluid channel. During operation, the liquid path system can form a negative pressure in the negative pressure tank by operating the waste liquid pump, and the negative pressure makes the sample loaded from the sampling needle to the flow chamber through the communication between the negative pressure tank and the flow chamber; the sheath liquid bottle is configured to provide the 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; and the optical signal collection lens is configured to transmit and collect the light beam. The flow cytometer improves the loading method, simplifies the operation procedure, and realizes continuous loading compared with the prior art. BRIEF DESCRIPTION OF DRAWINGS

[0050] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present disclosure, and therefore should not be regarded as a limitation on the scope, and other related drawings can also be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0051] Fig. 1 is a structural schematic diagram of a liquid path system provided in an embodiment of the present disclosure;

[0052] Fig. 2 is a schematic diagram of a cleaning channel of a swab provided in an embodiment of the present disclosure;

[0053] Fig. 3 is a structural schematic diagram of an optical signal collection lens provided in an embodiment of the present disclosure;

[0054] Fig. 4 is a working schematic diagram of the optical signal collection lens provided in an embodiment of the present disclosure;

[0055] FIG. 5 is a structural schematic diagram of a light signal collection lens provided in an embodiment of the present disclosure;

[0056] FIG. 6 is a working schematic diagram of an optical system provided in an embodiment of the present disclosure;

[0057] FIG. 7 is a cross-sectional schematic diagram of a flow chamber provided in an embodiment of the present disclosure;

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

[0059] Legend: 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 path channel; 123 - second flow path channel; 130 - damping device; 140 - first pressure sensor; 190 - first pipeline; 200 - sheath liquid pump; 240 - light source; 245 - forward light beam; 246 - lateral 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 - collection 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 - de-pulsation device; 20 - cleaning liquid pump; 30 - cleaning liquid bottle; 40 - second pressure sensor; 50 - waste liquid bottle; 60 - sixth valve; 70 - flow meter; 80 - one-way valve; 90 - air inlet filter; 1100 - liquid path system; 1200 - optical system; 2000 - sample. DETAILED DESCRIPTION

[0060] In order to make the objects, technical solutions and advantages of the embodiments of the present disclosure clearer, the technical solutions in the embodiments of the present disclosure will be described clearly and completely below with reference to the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are some but not all of the embodiments of the present disclosure. The components of the embodiments of the present disclosure described and shown in the drawings herein can be arranged and designed in various different configurations.

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

[0062] It should be noted that similar reference numbers and letters represent similar items in the following drawings, so once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0063] In the description of the present disclosure, it should be noted that if the terms "upper", "lower", "inner", "outer" and the like indicate the orientation or positional relationship shown in the drawings, or the orientation or positional relationship when the product is used, it is only for the convenience of describing the present disclosure and simplifying the description, and does not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present disclosure.

[0064] In addition, if the terms "first", "second" and the like appear, they are only used to distinguish the description and cannot be understood as indicating or implying relative importance.

[0065] It should be noted that the features in the embodiments of the present disclosure can be combined with each other without conflict.

[0066] As mentioned in the background, flow cytometry is usually configured for analysis and statistics of cells or particles in a sample. Flow cytometry mainly consists of a liquid system, an optical system, and a data analysis system. The liquid system, as one of the important components of the flow cytometer, transports the sample containing cells or particles together with the sheath liquid to the flow chamber, focuses the cells or particles using the principle of fluid dynamics, and makes the cells or particles pass through the detection area of the flow chamber one by one. Stable sample flow is the guarantee for subsequent signal detection and is one of the core technologies of the entire instrument.

[0067] The mainstream flow cytometer on the current market mostly has an absolute counting function. Absolute volume counting is achieved by measuring the volume of the measured sample, and then dividing the number of measured cells by the volume of the measured sample to obtain the concentration information of the measured sample, thereby achieving absolute counting. The scheme of absolute volume counting is mainly realized by a quantitative pump that can quantitatively transport the volume of the sample, such as a syringe pump, a plunger pump, and a peristaltic pump. The connection of the pipeline, sample suction, sample loading, and cleaning are relatively complex, which can also cause problems such as poor consistency of detection results, high contamination rate, etc. The syringe pump and the plunger pump cannot continuously load the sample, and need to first suck the sample and then load the sample, which is a complex operation procedure.

[0068] Therefore, referring to FIGS. 1 and 2, the flow cytometer and the control method thereof provided in the embodiments of the present disclosure can solve the problem, which will be described in detail as follows.

[0069] The flow cytometer provided in the embodiments includes a liquid path system 1100 and an optical system 1200, wherein the liquid path system 1100 is mainly configured to transport liquids such as the sample 2000, sheath liquid, cleaning liquid, and waste liquid, and the optical system 1200 is configured to form a specific light beam by irradiating the sample 2000 with a light source and then transmit and collect the light beam by a light signal collection lens 250. The light source can be a laser 240.

[0070] Next, referring to FIG. 1, the liquid path system 1100 in the embodiments will be described in detail. The liquid path 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 the like. The sampling needle 400, the sheath liquid bottle 700, and the negative pressure tank 100 are in communication with the flow chamber 300, and the sampling needle 400 and the sheath liquid bottle 700 are configured to deliver the sample 2000 and the sheath liquid to the flow chamber 300, respectively. The sheath liquid pump 200 is in communication 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 in communication with 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 as to facilitate the negative pressure tank 100 to provide negative pressure to the flow chamber 300. The flow chamber 300 is configured to accommodate the sample 2000 and the sheath liquid for detection in cooperation with the optical system 1200.

[0071] Referring to FIGS. 1 and 7, in order to facilitate the collection of waste liquid, the liquid path system 1100 further includes a waste liquid bottle 50. In addition, the liquid path system 1100 further includes a sample delivery needle 350, which is located in the flow chamber 300 and is in communication with the sampling needle 400.

[0072] It is worth noting that, during operation, the pressure in the negative pressure tank 100 can be dynamically maintained at a target pressure value by controlling the waste liquid pump 500, and then the flow rate of the sample 2000 can be adjusted by adjusting the sheath liquid flow rate, instead of dynamically maintaining the pressure difference between the two ends of the flow chamber 300.

[0073] In addition, the sheath liquid flow rate can be directly changed by changing the rotating speed of the sheath liquid pump 200 to change the sample loading speed, without setting a step flow valve to control the sheath liquid flow rate, and a greater flow rate adjustment range can be obtained.

[0074] The negative pressure tank 100 comprises a negative pressure tank body 120 and a plurality of control valves 110, which can be solenoid valves. The plurality of control valves are arranged on the negative pressure tank body 120. In this embodiment, the number of 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 the 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 the advantages of high integration and small size are achieved.

[0076] The first valve 111 is configured to control the conduction or blockage between the waste liquid pump 500 and the flow chamber 300, so that the conduction between the waste liquid pump 500 and the flow chamber 300 is facilitated when the first valve 111 is opened, thereby facilitating the loading process.

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

[0078] The third valve 113 is arranged on the first flow path 122, and the third valve 113 is configured to control the conduction or blockage of the first flow path 122, so as to control the conduction or blockage between the pressure relief port 121 and the negative pressure chamber inside the negative pressure tank body 120. The negative pressure tank 100 further comprises an air inlet filter 90, and the pressure relief port 121 is in communication with the air inlet filter 90.

[0079] In addition, in order to facilitate the monitoring of the pressure in the negative pressure tank 100, and at the same time, the pressure in the negative pressure tank 100 can be accurately controlled, and the pressure fluctuation is reduced, the negative pressure tank 100 further comprises a first pressure sensor 140 and a damping device 130. The negative pressure tank body 120 is provided with a second flow path 123, one end of the second flow path 123 is in communication with the inside of the negative pressure tank body 120, the first pressure sensor 140 is in communication with the other end of the second flow path 123, and the first pressure sensor 140 can be in communication with the negative pressure chamber inside the negative pressure tank body 120 through the second flow path 123. The first pressure sensor 140 facilitates the monitoring of the pressure in the negative pressure tank 100.

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

[0081] The damping device 130 is arranged in the negative pressure tank body 120, and the damping device 130 and the negative pressure tank body 120 are in communication, that is, the damping device 130 and the negative pressure chamber in the negative pressure tank body 120 are in communication.

[0082] The damping device 130 can be an air inlet damping pipe, a throttle valve, a speed regulating valve, or a filter, and the damping device 130 is in communication with the atmosphere, and is equivalent to a pressure stabilizing device or a buffer, etc. The damping device 130 is convenient for accurately controlling the pressure in the negative pressure tank 100 and effectively reducing the pressure fluctuation.

[0083] It is worth noting that when the pressure in the negative pressure tank 100 fluctuates, especially when the output fluctuation of the waste liquid pump 500 causes the pressure fluctuation to be 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 as a bypass of the negative pressure output end can make the pressure fluctuation of the negative pressure output end more balanced, thereby providing protection for the stable detection of the subsequent signal.

[0084] In addition, it needs to be explained that the negative pressure tank 100 can realize real-time emptying of the liquid in the negative pressure tank 100, and there is no need to store waste liquid, and thus there is no need to arrange a liquid level sensor.

[0085] Therefore, the waste liquid pump 500 in the embodiment can be a diaphragm pump with low precision and low cost, and can also accurately control the pressure in the negative pressure tank 100, thereby achieving the effect of smaller output pressure fluctuation.

[0086] Please refer to FIG. 2 in combination with FIG. 1. The liquid path system 1100 further comprises a swab 610, a swab pump 620, and a sheath liquid bottle 700. The swab 610 is provided with a cleaning channel 611. The swab pump 620 and one end of the cleaning channel 611 are in communication. The other end of the cleaning channel 611 and the sheath liquid bottle 700 are in communication. The sheath liquid pump 200 is arranged on a pipeline between the sheath liquid bottle 700 and the other end of the cleaning channel 611.

[0087] The sampling needle 400 and the cleaning channel 611 can slide relative to each other, so as to clean the sampling needle 400. Here, the sampling needle 400 and the cleaning channel 611 can slide relative to each other can be understood as that the sampling needle 400 is stationary, and the swab 610 moves downward, or that the swab 610 remains stationary, and the sampling needle 400 moves upward, which is equivalent to the retraction process of the sampling needle 400.

[0088] In the embodiment, the cleaning operation is preferably performed in a manner that the sampling needle 400 is stationary and the swab 610 moves, so as to shorten the extension length of the sampling channel as much as possible and facilitate the reduction of the fixed volume.

[0089] As shown in FIG. 2, the cleaning channel 611 has a main channel and first and second sub-channels 6111 and 6112 communicating with the main channel, the sampling needle 400 and the main channel of the cleaning channel 611 are relatively slidable, the first sub-channel 6111 and the swab pump 620 are connected by a pipeline, the liquid circuit further comprises a sixth valve 60 (which can be an electromagnetic valve), the sixth valve 60 is arranged on the pipeline between the first sub-channel 6111 and the swab pump 620, the second sub-channel 6112 is connected with the sheath liquid pump 200, and 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 pipeline between the sheath liquid pump 200 and the second sub-channel 6112 to be conducted or blocked.

[0090] In addition, in the embodiment, the liquid circuit system 1100 further comprises a degasser 800 and a first pipeline 190, the sheath liquid bottle 700, the sheath liquid pump 200, the degasser 800 and the flow chamber 300 are sequentially arranged on the first pipeline 190, the degasser 800 can remove air in the sheath liquid, solve the influence of air bubbles on the liquid flow, improve the stability of the liquid flow, improve the detection precision, and expand the adaptability of the flow cytometer to different use environments.

[0091] Specifically, the liquid circuit system 1100 further comprises a first filter 900 and a pulse elimination device 10, the sheath liquid bottle 700, the sheath liquid pump 200, the pulse elimination device 10, the first filter 900, the degasser 800 and the flow chamber 300 are sequentially arranged on the first pipeline 190. In the working process, the main function of the pulse elimination device 10 is to eliminate the pulse generated when the sheath liquid pump 200 works, so as to ensure the transmission stability of the liquid circuit system.

[0092] The fifth valve 115 is configured to simultaneously connect the degasser 800 and the flow chamber 300, the fifth valve 115 is also arranged on the first pipeline 190, and the fifth valve 115 can control the conduction or blockage 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 pipeline between the sheath liquid pump 200 and the second sub-channel 6112 to be conducted or blocked.

[0094] Meanwhile, the liquid path system 1100 further comprises a second filter (not shown in the figure), which can be arranged on the first pipeline 190 between the sheath liquid pump 200 and the sheath liquid bottle 700. The sheath liquid pump 200 can be 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 pulse elimination device 10, the degasser 800 and the flow chamber 300 can be arranged in sequence on the first pipeline 190.

[0096] In some embodiments, the liquid path system 1100 can comprise a second filter, and there is no first filter 900 and pulse elimination device 10.

[0097] Alternatively, the liquid path system 1100 comprises a first filter 900 and a pulse elimination device 10, and there is no second filter. At this time, the sheath liquid pump 200 can be a peristaltic pump.

[0098] In order to facilitate the cleaning process of the liquid path, the liquid path system 1100 further comprises a cleaning liquid pump 20 and a cleaning liquid bottle 30. One end (which can be understood as an inlet end) of the cleaning liquid pump 20 is in communication with the cleaning liquid bottle 30, and the other end (which can be understood as an outlet end) of the cleaning liquid pump 20 is in communication with the flow chamber 300.

[0099] The cleaning liquid pump 20 can deliver the cleaning liquid in the cleaning liquid bottle 30 to the flow chamber 300. Meanwhile, the cleaning liquid can enter the sample delivery needle, flow through the flow meter 70 and the sample needle 400, and complete the cleaning of the sample channel and the flow chamber 300. One of the waste liquids 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 waste liquid can enter the waste liquid bottle 50 through the sample needle 400, the first sub-channel 6111 and the swab pump 620.

[0100] In addition, the liquid path system 1100 further comprises a second pressure sensor 40 and a flow meter 70. The flow meter 70 is arranged on the pipeline between the sample needle 400 and the flow chamber 300. Specifically, the flow meter 70 can be arranged on the pipeline between the sample needle 400 and the sample delivery needle. 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 path system 1100 further comprises 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 on the third pipeline, and can prevent backflow.

[0102] In addition, in the embodiment, the second valve 112 is configured to connect the pipeline between the sixth valve 60 and the swab pump 620 and the flow chamber 300 at the same time, and the second valve 112 is configured to connect the pipeline between the sixth valve 60 and the swab pump 620 and the flow chamber 300 at the same time. The pipeline is turned on or off, so that in the case that the third valve 113 and the first valve 111 are opened, the external air enters the negative pressure tank 100, and the liquid in the flow chamber 300 flows through the second valve 112, the swab pump 620 and the waste liquid bottle 50 in turn, so as to achieve the purpose of emptying the flow chamber 300.

[0103] Referring to FIGS. 3-6, the embodiments of the present disclosure provide a light signal collection lens 250 configured to be a flow cytometer or a sample analysis device, the light signal collection lens 250 comprising a first transmission part 260, a second transmission part 270 and a collection part 280 arranged in sequence and spaced apart;

[0104] 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 is configured to receive irradiation of laser and generate a divergent light beam. The first transmission part 260 is formed with a baffle surface 262 on the side close to the second transmission part 270. The baffle surface 262 is configured to correct 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] In the working process of the light signal collection lens 250, the sample 2000 can be transported through the fluid channel 261, and the sample 2000 can receive laser irradiation at the same time to generate a divergent light beam. Then the divergent light beam is transmitted from the first transmission part 260 to the second transmission part 270 and the collection part 280. The baffle surface 262 is configured to correct the spherical aberration of the light beam transmitted by the first transmission part 260, thereby improving the transmission and collection effect of the light beam. It should be noted that the first transmission part 260 can be understood as a flow cell 310 on the flow chamber 300 in FIG. 7, 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 in a wave-shaped concave shape. The wave refers to the fact that the surface of the baffle surface 262 is in an arc shape with inconsistent curvature, which can also be understood as the fact that different positions on the baffle surface 262 have fluctuations. The concave refers to the fact that the two end positions 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 end portions of the baffle surface 262 in the extension direction of the fluid channel 261. The light 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 reflects 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. Moreover, when the light source 240 irradiates the sample 2000, the forward light beam 245 and the lateral light beam 246 are generated simultaneously, and the light signal collection lens 250 is configured to collect the lateral light beam 246.

[0108] In addition, in order to make the face type of the baffle surface 262 more accurate, the face type calculation equation of the baffle surface 262 can be defined as: z(h) = h2 / R{1+[1-(1+k)h2 / R2]1 / 2}+A2h2+A4h4+A6h6+A8h8+A10h10+…+Anhn, through the calculation formula, the specific position of each point on the baffle surface 262 is determined, which can improve the processing convenience and facilitate repeated processing.

[0109] Wherein, h is the distance between each point on the baffle surface 262 and the optical axis, R is the radius of curvature, k is the quadratic surface constant, A2, A4, A6, A8, A10…An is the high-order surface coefficient.

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

[0111] Moreover, according to multiple actual experiments, the specific values of each parameter can be obtained by backstepping from the obtained light signal effect. As preferred, 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 the larger the value of n in An is, the smaller the specific parameter is, so when n reaches a certain value, its influence on the final calculation of z(h) is quite limited. Under normal calculation conditions, n=10 can be taken, that is, the preferred face type calculation equation of the baffle surface 262 is: z(h) = h2 / R{1+[1-(1+k)h2 / R2]1 / 2}+A2h2+A4h4+A6h6+A8h8+A10h10.

[0112] Referring to FIGS. 3-5, the first transmission part 260 further comprises a refracting surface 263, which is a plane and located on the side of the fluid channel 261 away from the second transmission part 270.

[0113] When the divergent light beam is generated, the divergent light beam will first pass through the side wall of the fluid channel 261, and then be transmitted to the refracting surface 263, and continue to be transmitted in the direction away from the fluid channel 261 after being refracted by the refracting surface 263.

[0114] Referring to FIGS. 3-5, the first transmission portion 260 further comprises a reflecting surface 264, which is arc-shaped and located on the side of the refracting surface 263 away from the second transmission portion 270. That is, the refracting surface 263 is located between the reflecting surface 264 and the fluid channel 261.

[0115] In operation, the reflecting surface 264 is configured to receive the light beam transmitted through the refracting surface 263 and then reflect it back to the refracting surface 263, so as to move it towards the direction close to the second transmission portion 270, thereby playing a role of reflecting the light beam.

[0116] Specifically, in order to improve the reflecting effect, the surface of the reflecting surface 264 can be coated with a reflecting film 265, which can be a dielectric film or a metal film. The dielectric film can be composed of multiple layers of oxide materials with high and low refractive indexes, and the overall material thickness is about 0.1 nm-10 nm, and the specific thickness is not limited. The light beam is folded back multiple times after passing through the multiple layers, thereby reflecting most of the light beam and improving the reflectivity, so that the actual reflectivity can reach more than 99%. Although the reflectivity is also limited by the incident angle, by arranging the refracting surface 263 on the side of the reflecting surface 264 close to the fluid channel 261, the angle of the light beam incident to the reflecting surface 264 can be adjusted, so as to improve the collection efficiency of the light beam reflected by the reflecting surface 264 and reduce the light loss. The metal film is generally composed of metals such as gold, silver, and aluminum. Unlike individual metal materials, the reflectivity of different wave bands is inconsistent. In the visible light wave 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, the transverse edge of the reflecting surface 264 is located on the extension line of the diagonal of the same rectangular cross section of the fluid channel 261, and preferably the length of the rectangular cross section of the fluid channel 261 is between 100 um and 500 um. The 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, both of which are within the range of 100 um-500 um. The lower the lower limit of the length, the narrower the channel, which will cause too large flow resistance, resulting in too large pressure difference between the two ends, and thus too large negative pressure of the negative pressure tank 100. The higher the upper limit of the length, the wider the channel, which is not conducive to compressing the core flow and will cause too large flow rate of the sheath fluid, and too large consumption of the sheath fluid will also cause negative effects such as cost increase. By limiting the upper and lower limits of the length within a reasonable range, the width of the channel is moderate, the flow resistance is not too large, the pressure difference between the two 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 too large flow rate of the sheath fluid and cost increase.

[0118] Referring to FIGS. 3 and 5, the second transmission portion 270 is a convex lens.

[0119] In this embodiment, the second transmission part 270 is a three-lens cemented lens, and includes a first sub-lens 271, a second sub-lens 272 and a third sub-lens 273 arranged in sequence in the direction from the collection part 280 to the first transmission part 260.

[0120] Of course, alternatively, the second transmission part 270 can also be a single lens, a double-lens cemented lens, a four-lens cemented lens, a five-lens cemented lens, etc., and the specific number of sub-lenses therein is not limited.

[0121] Referring to FIGS. 3-5, the first sub-lens 271 is arc-shaped, with the middle position of the first sub-lens 271 closer to the collection part 280 than the two ends; and, the third sub-lens 273 can also be arc-shaped, with the middle position of the third sub-lens 273 closer to the first transmission part 260 than the two ends; while the second sub-lens 272 is a convex lens, with the two sides of the second sub-lens 272 cemented with the first sub-lens 271 and the third sub-lens 273, respectively.

[0122] In the process of beam propagation, when the light beam passes through the baffle surface 262, it enters the air part between the first transmission part 260 and the second transmission part 270, then reaches the third sub-lens 273 and successively passes through the second sub-lens 272 and the first sub-lens 271 before being emitted, and then converges to the collection part 280, forming an energy-concentrated light spot.

[0123] It is worth noting that the collection part 280 generally places a detector, such as a silicon-based photodetector, such as a CCD, a PD, an APD or a PMT, and can also place other functional optical elements, including but not limited to an aperture, a pinhole or a fiber bundle, configured to control the transmission of light.

[0124] In addition, the optical signal collection lens 250 adopts a folded structure, which adds a mirror system to the traditional refractive material. Since it is difficult to correct the chromatic aberration of the refractive material in a wide wavelength range, 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 is free of chromatic aberration in any desired bandwidth range, which also promotes the simplification of the structure and the reduction of the number of lenses. Therefore, by adding a mirror system to the traditional refractive material, the difficulty of chromatic aberration correction is reduced, and the lens structure is simplified.

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

[0126] The light signal collection lens 250 introduces a spherical reflecting surface 264, and the curvature center position is optimized to a non-spherical baffle surface 262 to correct spherical aberration, and the baffle surface 262 acts as a diaphragm, so that the system has a large uniform image field. Because the structure of the flow stream cavity is highly transparent, the material is generally made of optical glass, so the part of the optical lens can be made into an integrated structure with the liquid flow cavity, such as the first transmission part 260, so that the optical material composed of the reflecting surface 263 and the reflecting surface 264 is bonded together by optical gel or molecular bonding. If the fluid channel 261 is made of the same material, the reflecting surface 263 is not needed, and the reflecting surface 264 and the baffle surface 262 are directly optically processed. Specifically, CNC grinding or other processing methods can be used to optimize the reflecting surface 263 and the baffle surface 262 to a reasonable surface type. By separating the reflecting surface 263 and the baffle surface 262 of the optical lens from the fluid channel 261, the processing yield can be guaranteed. The reflecting surface 264 can be spherical, parabolic or other aspherical surfaces, etc. Considering the difficulty of processing and the sensitivity of manufacturing errors, a spherical surface is used in this embodiment, and the baffle surface 262 is used to correct the spherical aberration introduced by the reflecting surface 264. The remaining aberration is the change of spherical aberration with wavelength or some high-order astigmatism and the oblique ray spherical aberration introduced by off-axis light. Although the reflecting surface 264 and the liquid flow cavity of the same material can reduce the chromatic aberration to a low level, the change of refractive index of the material will also produce a certain chromatic aberration or secondary spectrum in the wide spectral domain because the light first exits from the aqueous solution to the glass and then from the glass to the air. The light signal collection lens 250 sets the 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 indexes to correct chromatic aberration or secondary spectrum. The number and thickness of the flint glass and the crown glass are not limited, and they can be double-cemented lenses or triple-cemented lenses.

[0127] The second transmission part 270 is a triple-cemented lens, which is composed of two pieces of flint glass with high refractive index and one piece of double-convex crown glass with low refractive index. It is configured to correct the remaining chromatic aberration and coma aberration, 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, which has a certain degree of freedom of choice. The center low refractive index crown glass generally uses K9 or BK7 glass, which has the characteristics of low cost and stable performance.

[0128] The middle of the first transmission part 260 and the second transmission part 270 is air, which acts as a spacing. Because the optical system 1200 composed of the reflecting surface and the baffle surface 262 is in an infinite conjugate form, the air spacing distance in the middle of the two parts has no limit requirement, and corresponding optical elements or other devices can be inserted in the middle, which increases the degree of freedom in optical and structural design.

[0129] It is worth noting that the effect that can be achieved by the optical signal collection lens 250 includes: the numerical aperture reaches 1.21, the field of view reaches 500um, the work distance is greater than 2mm, the RMS radius of the focused light plate is less than 50um, the wavelength covers 400nm-900nm, the magnification is 9 times, and by limiting each parameter, the 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 the embodiment can refer to Table 1 (unit: mm) as follows:

[0131] Table 1

[0132] It is worth noting that the radius, interval and refractive index parameters in Table 1 are mostly range values, and the preferred value is usually the middle value close to the range value, but the specific selection value in the design process remains within the allowed range.

[0133] The embodiment provides a control method of a flow cytometer, which is applied to control the flow cytometer, and specifically includes the following steps:

[0134] Step S100: control the formation of negative pressure in the negative pressure tank 100, and sample the sample 2000 on the flow chamber 300 through the sampling needle 400;

[0135] Step S200: control the sheath liquid pump 200 to deliver sheath liquid to the flow chamber 300, so that the sheath liquid wraps the sample 2000.

[0136] Specifically, step S100 includes:

[0137] Step S110: control the constant negative pressure formed in the negative pressure tank 100;

[0138] Step S120: control the sampling needle 400 to aspirate the sample 2000 and sample it to the flow chamber 300.

[0139] It should be noted that the above steps S100 and S200 can be operated in the order of S100 first and S200 second, or in the order of S100 second and S200 first, or simultaneously. The specific operation order can be determined according to the actual collection situation.

[0140] Further, in order to further ensure the accuracy and stability of the whole operation, the pressure of the negative pressure tank 100 can be kept constant during the operation, and the adjustment of the sample 2000 flow is realized by adjusting the sheath liquid flow. The advantage of this control method is that it can ensure the stability of the nuclear flow, which is reflected in the width, position and flow rate of the nuclear flow. In the normal loading state, the position of the nuclear flow changes less, unless the sample 2000 flow or the sheath liquid flow fluctuates greatly, the width and flow rate of the nuclear flow are mainly determined by the sample 2000 flow. 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 sample 2000 flow, the sample 2000 flow actually depends on the pressure in the flow chamber 300, which can also be understood as controlling the stability of the sample 2000 flow by the sheath liquid pump 200, which is to control the stability of the pressure in the flow chamber 300 first, and the stability of the flow in the flow cell 310 is naturally brought about by the stability of the pressure in the flow chamber 300 and the negative pressure tank 100.

[0141] The step S110 comprises:

[0142] Step S111: obtaining the real-time pressure in the negative pressure tank 100;

[0143] Step S112: controlling the waste liquid pump 500 to perform PID adjustment on the pressure in the negative pressure tank 100. The step S200 comprises:

[0144] Step S210: controlling the sheath liquid pump 200 to perform PID control on the flow of the sample 2000 entering the sampling needle 400.

[0145] Specifically, the first pressure sensor 140 can feed back the closed-loop PID control of the waste liquid pump 500 (PID control is a control system based on proportional, integral and differential control methods, which can eliminate errors by adjusting the real-time data of the controlled object and the given value, and achieving the control target by using proportional, integral and differential control methods), so that the negative pressure tank 100 maintains constant negative pressure; Similarly, the flow meter 70 feeds back the closed-loop PID control of the sheath liquid pump 200 to control and adjust the loading speed. Moreover, the two PID control methods are relatively independent, and the target value and the feedback value belong to the same attribute value, without the need for indirect conversion, thereby having the characteristics of simpler control and faster real-time feedback. Moreover, it can avoid the structure of double light sources and double channels in the prior art, so that the structure is also simplified.

[0146] It is worth noting that the existing technology is to measure the laser delay or sample flow rate through a double light source device and a double scattering channel, and then derive the pressure difference reference, and indirectly maintain the overall volume flow rate and sample flow rate stable by controlling the pressure difference between the flow channels of the flow chamber. The stability of the overall volume flow rate controlled by this adjustment method is not equal to the stability of the sample flow rate, because the total volume passing through the flow channel is composed of sheath liquid and sample. Although the overall volume flow rate is kept stable, the ratio of sample flow rate to sheath flow rate will change, which will affect the detection result accuracy. Moreover, 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 complex, the real-time feedback slower, and the structure more complex.

[0147] Please refer to FIG. 8. According to the above control method, the target pressure value of the negative pressure tank 100 is P1, which is controlled to be a value between -60 Kpa and -10 Kpa. The PID is used to adjust the pressure in the negative pressure tank 100, that is, the pressure in the negative pressure tank 100 is dynamically and constantly maintained at P1. The lower the target pressure value of the negative pressure tank 100, the more time it takes to establish negative pressure in the negative pressure tank 100, and the longer the preparation time of the flow cytometer, 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, and thus increase the fluctuation of the sample flow and the nuclear 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 will inevitably be lower. The target pressure value of the PID control of the pressure in the negative pressure tank 100 is set in this range, which comprehensively considers the factors such as the preparation time to establish negative pressure before sampling, the influence of pressure fluctuation on adjustment, and the adjustment accuracy control of the waste liquid pump 500.

[0148] The flow rate of the sample 2000 entering the sampling needle 400 is PID controlled by the sheath liquid pump 200, and the flow rate value is set as the target value. During the detection process of the sample 2000, the flow rate value is set as Q1, wherein 5ul / min≤Q1≤200ul / min. The interval setting of the sample 2000 flow rate is mainly determined by the speed of the sampling detection and the accuracy of the sampling detection. When the sample 2000 flow rate is too low, the collection time will be lengthened, and when the sample 2000 flow rate is too high, the nuclear flow size will be too large, which will reduce the detection accuracy.

[0149] Specifically, the flow rate of the sheath liquid flowing to the flow chamber 300 can be Q2, the flow resistance of the sample 2000 flowing through the sampling channel section can be R1, the flow resistance of the fluid channel in the flow cell can be R3, and the formula Q1 = (-P1-Q2*R3) / (R1+R3) can be combined, wherein R1, R3, and P1 are constant values. That is, by adjusting Q2, the purpose of adjusting Q1 can be achieved.

[0150] In the sample loading detection process, by maintaining the constant pressure P1 of the negative pressure tank 100, and by PID control of the sample 2000 flow rate Q1 by the sheath liquid pump 200, the stability of the nuclear flow size, position and liquid flow can be maintained, and the accuracy of the detection result can be ensured. In addition, by adjusting and changing the sheath liquid flow rate Q2 by the sheath liquid pump 200, the sample flow rate Q1 can be changed, and different sample loading detection speed requirements can be achieved.

[0151] Step 400: Control the light source to irradiate the sample 2000 and generate a light beam, and make the light signal collection lens 250 transmit and collect the light beam.

[0152] Of course, in order to improve the cleanliness of the sampling needle 400 and avoid the negative impact of low cleanliness of the sampling needle 400 on the sampling operation process, the control method can further include step S500: controlling the sheath liquid bottle 700 to deliver sheath liquid to the cleaning channel 611 to clean the outer wall of the sampling needle 400.

[0153] In addition, in order to realize stable sample loading operation, the control method can start the operation of the waste liquid pump 500 and the sheath liquid pump 200, and simultaneously open the first valve 111 and the fifth valve 115. The waste liquid pump 500 provides negative pressure, so that the sampling needle can suck the sample into the flow chamber 300. At the same time, the sheath liquid pump 200 delivers sheath liquid to the flow chamber 300, so that the sheath liquid wraps the sample 2000 liquid to form a laminar flow for optical detection.

[0154] Then the waste liquid enters the negative pressure tank body 120 inside the negative pressure tank 100 through the first valve 111, and then enters the waste liquid bottle 50 through the waste liquid pump 500.

[0155] As can be seen, in the embodiment, the negative pressure source is formed by the cooperation of the negative pressure tank 100 and the waste liquid pump 500, which can realize continuous sample loading measurement process, and replaces the injection pump and plunger pump used in the prior art for sample loading process, simplifies the pipeline connection, and reduces the liquid path volume.

[0156] 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 start-up time of the sample loading, and improve the detection efficiency of the flow cytometer.

[0157] In order to facilitate the cleaning of the outer wall of the sampling needle 400, the control method provided in the embodiment further includes:

[0158] The swab pump 620 and the sheath liquid pump 200 are controlled to work, so that the sheath liquid in the sheath liquid bottle 700 flows through the cleaning channel 611 and the outer wall of the sampling needle 400, and the sampling needle 400 and the swab 610 are controlled to move relatively to clean the outer wall of the sampling needle 400.

[0159] Specifically, by starting the swab pump 620 and the sheath liquid pump 200, and opening the fourth valve 114 and the sixth valve 60, the sheath liquid in the sheath liquid bottle 700 sequentially passes through the sheath liquid pump 200, the pulse removing device 10, the bypass of the first filter 900, and the fourth valve 114, and then enters the main channel of the swab 610, and the sampling needle 400 and the cleaning channel 611 slide relatively to clean the outer wall of the sampling needle 400, and the waste liquid generated in the cleaning process flows into the waste liquid bottle 50 through the sixth valve 60 and the swab pump 620.

[0160] It should be noted that, after each completion of the sample 2000 loading process, the cleaning process of the outer wall of the sampling needle 400 is required, and since the sheath liquid flows through the first filter 900, the cleaning process of the outer wall of the sampling needle 400 is also completed.

[0161] In order to facilitate the cleaning of the inner wall of the sampling needle 400, the control method further comprises:

[0162] The swab pump 620 and the sheath liquid pump 200 are controlled to work, so that the sheath liquid in the sheath liquid bottle 700 sequentially flows through the sheath liquid pump 200, the flow chamber 300, the internal sampling channel of the sampling needle 400, the cleaning channel 611, and the swab pump 620.

[0163] Specifically, by starting the swab pump 620 and the sheath liquid pump 200, and opening the fifth valve 115 and the sixth valve 60, the sheath liquid in the sheath liquid bottle 700 sequentially passes through the sheath liquid pump 200, the pulse removing device 10, the main channel of the first filter 900 (not the bypass of the first filter 900 mentioned above), the deaerator 800, and the fifth valve 115, and then enters the flow chamber 300 and the internal sampling channel of the sampling needle 400 to clean the inner wall of the sampling needle 400, and then the waste liquid enters the waste liquid bottle 50 through the first sub-channel 6111, the sixth valve 60, and the swab pump 620.

[0164] In addition, in order to facilitate the cleaning of the flow chamber 300 and the sampling channel of the sampling needle 400 by the cleaning liquid, the control method further comprises:

[0165] First, the cleaning liquid pump 20 is controlled to work to drive the cleaning liquid in the cleaning liquid bottle 30 to enter the flow chamber 300 and the internal sampling channel of the sampling needle 400. Specifically, the cleaning liquid pump 20 is started first to transport the cleaning liquid in the cleaning liquid bottle 30 to the flow chamber 300 and the internal sampling channel of the sampling needle 400.

[0166] After the delivery of the cleaning liquid is completed, 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 (which can be set according to specific conditions) to improve the cleaning effect.

[0167] Then, the waste liquid discharge process is performed, in which one way of discharging the waste liquid is as follows: by controlling the waste liquid pump 500 to work, at the same time, the first valve 111 is opened, so that the negative pressure tank 100 body of the negative pressure tank 100 forms a negative pressure to discharge the waste liquid in the flow chamber 300 and the internal sampling channel of the sampling needle 400, and the waste liquid enters the waste liquid bottle 50 in sequence through the first valve 111, the negative pressure tank 100 and the waste liquid pump 500.

[0168] Another way of discharging the waste liquid is as follows: by controlling the swab pump 620 to work, the sixth valve 60 is opened to discharge the waste liquid in the flow chamber 300 and the internal sampling channel of the sampling needle 400, and the waste liquid enters the waste liquid bottle 50 in sequence through the first sub-channel 6111, the sixth valve 60 and the swab pump 620.

[0169] In addition, in order to realize the flow chamber 300 bubble removal process, the control method further comprises:

[0170] First, the flow chamber 300 is emptied, and by controlling the swab pump 620 to work, the 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.

[0171] Specifically, the swab pump 620 is started, the first valve 111, the second valve 112 and the third valve 113 are opened, the external air enters the negative pressure tank 100 body through the air inlet filter 90, the third valve 113, and then enters the flow chamber 300 through the first valve 111, and the liquid in the flow chamber 300 enters the waste liquid bottle 50 through the second valve 112 and the swab pump 620, realizing the process of emptying the flow chamber 300.

[0172] It should be noted that when it is necessary to discharge the flow chamber 300, the above-mentioned liquid path control method for emptying the flow chamber 300 can also be used.

[0173] Then, the process of filling the flow chamber 300 is performed, and specifically, the control method further comprises:

[0174] The waste liquid pump 500 and / or the sheath liquid pump 200 are controlled to work to make the sheath liquid pass through the flow chamber 300, the negative pressure tank 100 and the waste liquid pump 500 in sequence to fill the flow chamber 300.

[0175] Specifically, opening the fifth valve 115 and the first valve 111, the sheath liquid in the sheath liquid bottle 700 can flow into the flow chamber 300 through the sheath liquid pump 200, the first filter 900, the degasser 800 and the fifth valve 115 in sequence, and then flow into the waste liquid bottle 50 through the first valve 111, the inside of the negative pressure tank 100 and the waste liquid pump 500, thereby realizing the filling of the flow chamber 300 and completing the process of removing bubbles from the flow chamber 300.

[0176] It should be noted that when it is necessary to drain the flow chamber 300, the above-mentioned liquid path control method for filling the flow chamber 300 can also be used.

[0177] That is, in the embodiment, the step of controlling the swab pump 620 to work so that the external gas flows into the negative pressure tank 100, the flow chamber 300 and the swab pump 620 in sequence to empty the flow chamber 300 is the first step; the step of controlling the waste liquid pump 500 and / or the sheath liquid pump 200 to work so that the sheath liquid flows through the flow chamber 300, the negative pressure tank 100 and the waste liquid pump 500 in sequence to fill the flow chamber 300 is the second step. The control method further comprises that the first step is located before the second step, that is, the first step is performed first, and then the second step is completed to realize the process of removing bubbles from the flow chamber 300.

[0178] In order to realize the draining of the sampling channel, the control method further comprises:

[0179] Controlling the waste liquid pump 500 to work so that a negative pressure is formed in the negative pressure tank 100 to make the sampling needle 400 draw the draining liquid, the cleaning liquid or the air and flow through the flow chamber 300, the negative pressure tank 100, the waste liquid pump 500 and the waste liquid bottle 50 in sequence.

[0180] Specifically, starting the waste liquid pump 500 while opening the first valve 111 so that the sampling needle 400 draws the draining liquid, the cleaning liquid or the air and flows into the waste liquid bottle 50 through the flow chamber 300, the first valve 111, the inside of the negative pressure tank 100 and the waste liquid pump 500 in sequence.

[0181] In addition, it should be noted that the above control methods can be combined to complete the complex actions such as starting and shutting down the flow cytometer and deep cleaning.

[0182] For example, in the starting process of the flow cytometer, the first filter 900 bubble removal process can be performed at the same time, and the flow chamber 300 bubble removal process can also be performed, so as to facilitate the removal of bubbles from the liquid path and improve the detection stability.

[0183] During the shutdown of the flow cytometer, the flow chamber 300 cleaning process mentioned above can be performed. In addition, a deep cleaning can also be achieved.

[0184] In summary, the flow cytometer, optical signal collection lens and sample analysis device provided by the embodiment have at least the following advantages:

[0185] 1. The negative pressure is provided by the waste liquid pump 500, and the sheath liquid supply amount is controlled by the sheath liquid pump 200, so as to adjust the sample 2000 loading speed. The flow meter 70 is used to detect the sample 2000 flow, so as to realize absolute volume counting and continuous loading measurement. Compared with the injection pump and the plunger pump loading mode used in the prior art, the pipeline connection is simple, and the liquid path volume is greatly reduced.

[0186] 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 can accurately control the pressure in the negative pressure tank 100, effectively reducing the pressure fluctuation. In addition, the negative pressure tank 100 can realize real-time emptying of the liquid in the negative pressure tank 100, without the need to store waste liquid, so that a liquid level sensor does not need to be set. At the same time, the waste liquid pump 500 can select a diaphragm pump with lower precision and lower cost, which can also accurately control the pressure in the negative pressure tank 100, and realize the effect of smaller output pressure fluctuation.

[0187] 3. The de-pulsing device 10 can effectively remove the liquid flow pulsation generated by the sheath liquid pump 200, improve the stability of the liquid flow, and improve the detection accuracy.

[0188] 4. The cleaning liquid pump 20 can effectively clean the flow chamber 300 and the sampling channel, improve the consistency of the detection result, reduce the carryover pollution rate, and make the flow cytometer function more complete and the automation degree higher.

[0189] 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 buffered pressure of the negative pressure tank 100 can reduce the start-up time of the beginning of collection, and improve the detection efficiency of the flow cytometer.

[0190] 6. The air remover 800 is used to remove the air content in the sheath liquid, solve the influence of the bubbles on the liquid flow, improve the stability of the liquid flow, improve the detection accuracy, and expand the adaptability of the flow cytometer to different use environments.

[0191] The control method of the flow cytometer mentioned the prior art using the method of first suction and then sample loading, realizes the continuous sample loading process, and the operation and cleaning process are simple, solves the problems of complex pipeline connection, sample suction, sample loading and cleaning in the prior art, and causes poor consistency of detection results, high pollution rate and other problems.

[0192] The above is only a specific embodiment of the present disclosure, but the protection scope of the present disclosure is not limited thereto, and any person skilled in the art can easily think of changes or replacements within the technical range disclosed by the present disclosure, which should be covered within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be subject to the protection scope of the claims. Industrial applicability

[0193] The flow cytometer improves the sample loading method relative to the prior art, simplifies the operation procedure, and realizes continuous sample loading.

Claims

1. A flow cytometer, characterized by, The application relates to a liquid path system (1100) and an optical system (1200). The liquid path system (1100) comprises a sampling needle (400), a sheath liquid bottle (700) and a flow chamber (300), the sampling needle (400) and the sheath liquid bottle (700) are both communicated with the flow chamber (300) and are respectively configured to deliver a sample (2000) and sheath liquid to the flow chamber (300). The optical system (1200) comprises a light source (240) and a light signal collection lens (250), the light signal collection lens (250) comprises a fluid channel (261) communicated with the flow chamber (300), the light source (240) is configured to irradiate the sample (2000) flowing into the fluid channel (261) from the flow chamber (300), and the light signal collection lens is configured to collect and transmit a light beam generated by the sample (2000) after being irradiated. The liquid path system (1100) further comprises a negative pressure tank (100) and a waste liquid pump (500), the waste liquid pump (500) is communicated with the flow chamber (300), the negative pressure tank (100) is arranged between the waste liquid pump (500) and the flow chamber (300), and the waste liquid pump (500) is operated to form negative pressure in the negative pressure tank (100), so that the sample (2000) is transferred from the sampling needle (400) to the flow chamber (300) and into the fluid channel (261). The waste liquid pump (500) is controlled to dynamically maintain the pressure in the negative pressure tank (100) at a target pressure value.

2. The flow cytometer of claim 1, wherein, The negative pressure tank (100) comprises a first pressure sensor (140), the first pressure sensor (140) is configured to acquire a real-time pressure value in the negative pressure tank (100), the real-time pressure value is used as a feedback value, and a target pressure value is used as a target value, and the pressure in the negative pressure tank (100) is PID controlled by the waste liquid pump (500).

3. The flow cytometer of claim 2, wherein, The target pressure value is P1, wherein -60Kpa<=P1<=-10Kpa.

4. The flow cytometer of claim 2, wherein, The liquid path system (1100) further comprises a sheath liquid pump (200), the sheath liquid pump (200) is communicated between the sheath liquid bottle (700) and the flow chamber (300), and the sheath liquid pump (200) is controlled to adjust the sheath liquid supply amount to adjust the flow rate of the sample (2000) flowing to the flow chamber (300).

5. The flow cytometer according to any one of claims 1 to 4, wherein, The liquid path system (1100) further comprises a flow meter (70), 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 acquire a real-time flow rate value of the sample (2000) flowing to the flow chamber (300), the real-time flow rate value is used as a feedback value, a set flow rate value is used as a target value, and the flow rate of the sample (2000) entering the sampling needle (400) is PID controlled by the sheath liquid pump (200).

6. The flow cytometer of claim 5, wherein, ​ 7. The flow cytometer of claim 6, wherein, 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, wherein, The negative pressure tank (100) comprises a negative pressure tank body (120) and a damping device (130), the damping device (130) is arranged on the negative pressure tank body (120), and the damping device (130) and the negative pressure tank body (120) are in communication; 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 path channel (122) and a pressure relief port (121), one end of the first flow path channel (122) is in communication with the pressure relief port (121), and the other end of the first flow path channel (122) is configured to be in communication with the negative pressure tank body (120), and the pressure relief port (121) is configured to realize 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 on the first flow path channel (122) and is configured to realize the on-off of 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, wherein, The liquid path system (1100) further comprises a swab (610), and the swab (610) is provided with a cleaning channel (611); and the sampling needle (400) is in sliding fit with the cleaning channel (611). Preferably, the cleaning channel (611) is slidable relative to the sampling needle (400).

10. The flow cytometer of claim 9, wherein, 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 liquid bottle (700). And / or, the liquid path system (1100) further comprises a cleaning liquid pump (20) and a cleaning liquid bottle (30), one end of the cleaning liquid pump (20) is in communication with the cleaning liquid bottle (30), and the other end of the cleaning liquid pump (20) is in communication with the flow chamber (300).

11. The flow cytometer according to any one of claims 1 to 10, wherein, The liquid path system (1100) further comprises a waste liquid bottle (50), 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 path system (1100) further comprises a sheath liquid pump (200), a degasser (800) and a first pipeline (190), the sheath liquid bottle (700), the sheath liquid pump (200), the degasser (800) and the flow chamber (300) are sequentially arranged on the first pipeline (190); And / or, the liquid path system (1100) comprises a sheath liquid pump (200) and a de-pulsation device (10), one end of the de-pulsation device (10) is in communication with the sheath liquid pump (200), and the other end of the de-pulsation device (10) is in communication with the flow chamber (300).

12. The flow cytometer of any one of claims 1-11, wherein, The light signal collection lens (250) comprises a first transmission part (260), a second transmission part (270) and a collection part (280) arranged in sequence. The first transmission part (260) is provided with the 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 being irradiated by laser. The first transmission part (260) is formed with a baffle surface (262) on the side close to the second transmission part (270), and the baffle surface (262) is configured to correct spherical aberration of the lateral light beam (246).

13. The flow cytometer of claim 12, wherein, The baffle surface (262) is in a wave-shaped concave shape. The first transmission part (260) further comprises a reflecting surface (264), and the reflecting surface (264) is an arc surface and is located on the side of the fluid channel (261) away from the second transmission part (270). The first transmission part (260) further comprises a reflecting surface (264) and a refracting surface (263), and the refracting surface (263) is a plane and is located between the reflecting surface (264) and the fluid channel (261). The first transmission part (260) further comprises a reflecting surface (264), and the reflecting surface (264) is coated with a reflecting film (265), and the reflecting film (265) is a dielectric film or a metal film.

14. The flow cytometer of claim 13, wherein, The cross section of the fluid channel (261) is rectangular, and the transverse edge of the reflecting surface (264) is located on the extension line of the diagonal of the same rectangular cross section of 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 collection lens characterized by, Comprise: a first transmission part (260), a second transmission part (270) and a collection part (280) arranged in sequence; The first transmission part (260) is provided with the 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 being irradiated by laser. The first transmission part (260) is formed with a baffle surface (262) on the side close to the second transmission part (270), and the baffle surface (262) is configured to correct spherical aberration of the lateral light beam (246).

16. The light signal collection lens according to claim 15, characterized in that, The baffle surface (262) is in a wave-shaped concave shape.

17. The light signal collection lens of claim 15, wherein, The first transmission part (260) further comprises a reflecting surface (264), and the reflecting surface (264) is an arc surface and is located on the side of the fluid channel (261) away from the second transmission part (270).

18. The light signal collection lens according to claim 17, wherein, The first transmission part (260) further comprises a refracting surface (263), and the refracting surface (263) is a plane and is located between the reflecting surface (264) and the fluid channel (261). The reflecting surface (264) is coated with a reflecting film (265), and the reflecting film (265) is a dielectric film or a metal film.

19. The light signal collection lens according to any one of claims 15-18, wherein, The second transmission part (270) is a three-lens cemented lens, and comprises a first sub-lens (271), a second sub-lens (272) and a third sub-lens (273) arranged in sequence in the direction from the collection part (280) to the first transmission part (260).

20. The light signal collection lens of claim 19, wherein, The first sub-lens (271) is arc-shaped, and the middle position of the first sub-lens (271) is closer to the collection part (280) than the two 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) than the two ends.

21. A sample analysis apparatus, characterized by, A flow cytometer comprising the optical signal collection lens of any one of claims 1-14 or the light signal collection lens of any one of claims 15-20.

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