CN113125330B - Flow cytometry analyzer - Google Patents
Flow cytometry analyzer Download PDFInfo
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- CN113125330B CN113125330B CN202110332240.1A CN202110332240A CN113125330B CN 113125330 B CN113125330 B CN 113125330B CN 202110332240 A CN202110332240 A CN 202110332240A CN 113125330 B CN113125330 B CN 113125330B
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- pipeline
- waste liquid
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- liquid
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1404—Fluid conditioning in flow cytometers, e.g. flow cells; Supply; Control of flow
Abstract
The embodiment of the invention discloses a flow cytometry analyzer which comprises a reaction tank for reacting liquid to be analyzed, a flow chamber for analyzing the liquid to be analyzed after the reaction in the reaction tank, a liquid tank for temporarily storing waste liquid discharged from the flow chamber, a first pipeline, a second pipeline, a third pipeline and a three-way valve. The two ends of the first pipeline are respectively communicated with the flow chamber and the liquid tank, the two ends of the second pipeline are respectively communicated with one of the liquid tank and the three-way valve, the other two ports in the three-way valve are respectively used for being communicated with the positive pressure port and the normal pressure port, one end of the third pipeline is communicated with the liquid tank, and the other end of the third pipeline is used for being communicated with the waste liquid port. When the liquid tank receives the waste liquid, the three-way valve is communicated with the normal pressure port, so that the waste liquid stably flows into the liquid tank; when the waste liquid in the liquid tank is discharged, the three-way valve is communicated with the positive pressure port, so that the waste liquid enters the waste liquid interface through the third pipeline.
Description
Technical Field
The invention relates to the technical field of flow cytometry analysis, in particular to a flow cytometry analyzer.
Background
Flow cytometry, also known as sheath flow, is a technique used to detect, analyze and count cells.
In the prior art, a flow cytometry analyzer generally includes a reaction tank for reacting a sample, and a flow chamber for counting cells in the reacted sample, wherein the reaction tank is connected with one end of the flow chamber through a first pipeline to output a sample flow, and after counting in the flow chamber, waste liquid generated in the flow chamber is required to be discharged. At present, the waste liquid is treated by being communicated with a direct waste liquid interface through a pipeline, and by adopting the structure, pressure impact can be generated on a waste liquid pipe orifice when the waste liquid is discharged from other channels. Under the influence of this pressure impact, turbulence of the sample flow can result, thereby affecting the results of the analysis.
Disclosure of Invention
Accordingly, it is necessary to provide a flow cytometer capable of solving the problem of pressure impact on the waste liquid gate when the waste liquid is discharged.
A flow cytometry analyzer comprises a reaction tank for reacting liquid to be analyzed, a flow chamber for analyzing the liquid to be analyzed after the reaction in the reaction tank, a liquid tank for temporarily storing waste liquid discharged from the flow chamber, a first pipeline, a second pipeline, a third pipeline and a three-way valve;
two ends of the first pipeline are respectively communicated with the flow chamber and the liquid tank, two ends of the second pipeline are respectively communicated with one of the liquid tank and the three-way valve, the other two ports in the three-way valve are respectively used for communicating a positive pressure port and a normal pressure port, one end of the third pipeline is communicated with the liquid tank, and the other end of the third pipeline is used for communicating a waste liquid interface;
when the liquid tank receives the waste liquid, the three-way valve is communicated with the normal pressure port, so that the waste liquid stably flows into the liquid tank; when the waste liquid in the liquid tank is discharged, the three-way valve is communicated with the positive pressure port, so that the waste liquid enters the waste liquid interface through the third pipeline.
The flow cytometry provided by the embodiment of the invention is communicated with the normal pressure port through the three-way valve, so that the air pressure in the liquid tank is always unchanged, and the waste liquid flows into the liquid tank at a stable speed. And the waste liquid in the liquid tank enters a third pipeline and finally flows out from the waste liquid interface through the three-way valve and the positive pressure port. By adopting the structure, the impact of waste liquid on the pressure of a pipeline for transporting the waste liquid is avoided, thereby improving the accuracy of flow cytometry analysis.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Wherein:
fig. 1 is a schematic structural diagram of a flow cytometer provided in the first embodiment.
Fig. 2 is a schematic structural diagram of a flow cytometer provided in a second embodiment.
Fig. 3 is a schematic structural diagram of a flow cytometer provided in a third embodiment.
Fig. 4 is a schematic structural diagram of a flow cytometer provided in the fourth embodiment.
Reference numerals:
110-reaction tank, 120-flow chamber, 130-liquid tank, 140-three-way valve, 150-positive pressure port, 160-normal pressure port, 170-waste liquid interface, 180-injection component, 181-first injector, 182-second injector;
210-first line, 220-second line, 230-third line, 240-fourth line, 250-fifth line, 260-sixth line, 270-seventh line, 280-eighth line, 290-ninth line, 300-tenth line, 310-eleventh line;
410-first valve, 420-second valve, 430-third valve, 440-fourth valve, 450-fifth valve, 460-sixth valve, 470-seventh valve, 480-eighth valve, 490-ninth valve;
510-swab, 511-sampling needle, 512-wiper, 520-WBC pool.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that all directional indicators (such as up, down, left, right, front, and rear … …) in the embodiments of the present invention are merely used to explain the relative positional relationship, movement, etc. between the components in a specific posture (as shown in the drawings), and if the specific posture is changed, the directional indicators are correspondingly changed.
Furthermore, the description of "first," "second," etc. in this disclosure is for descriptive purposes only and is not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In addition, the technical solutions of the embodiments may be combined with each other, but it is necessary to base that the technical solutions can be realized by those skilled in the art, and when the technical solutions are contradictory or cannot be realized, the combination of the technical solutions should be considered to be absent and not within the scope of protection claimed in the present invention.
Referring to fig. 1, an embodiment of the present invention provides a flow cytometer including a reaction tank 110 for reacting a liquid to be analyzed, a flow chamber 120 for analyzing the liquid to be analyzed after reacting in the reaction tank 110, a liquid tank 130 for temporarily storing a waste liquid discharged from the flow chamber 120, a first pipeline 210, a second pipeline 220, a third pipeline 230, and a three-way valve 140. Two ends of the first pipeline 210 are respectively communicated with the flow chamber 120 and the liquid tank 130, two ends of the second pipeline 220 are respectively communicated with one of the liquid tank 130 and the three-way valve 140, the other two ports in the three-way valve 140 are respectively used for communicating the positive pressure port 150 and the normal pressure port 160, one end of the third pipeline 230 is communicated with the liquid tank 130, and the other end of the third pipeline 230 is used for communicating the waste liquid interface 170.
Specifically, when the liquid tank 130 receives the waste liquid, the three-way valve 140 communicates with the normal pressure port 160 so that the waste liquid stably flows into the liquid tank 130; when the waste liquid in the liquid tank 130 is discharged, the three-way valve 140 communicates with the positive pressure port 150 so that the waste liquid enters the waste liquid port 170 through the third pipe 230.
With the flow cytometer provided above, the three-way valve 140 is communicated with the normal pressure port 160, so that the air pressure in the liquid tank 130 is always unchanged, and thus the waste liquid flows into the liquid tank 130 at a stable speed. And then is communicated with the positive pressure port 150 through the three-way valve 140, so that the waste liquid in the liquid tank 130 enters the third pipeline 230 and finally flows out of the waste liquid interface 170. By adopting the structure, the impact of waste liquid on the pressure of a pipeline for transporting the waste liquid is avoided, thereby improving the accuracy of flow cytometry analysis.
The positive pressure port 150, i.e., a port having a pressure greater than normal pressure, may apply a certain force to the waste liquid when the positive pressure port 150 is in communication with the liquid tank 130. The atmospheric pressure port 160 is an atmospheric pressure. In addition, the positive pressure port 150 may be an output port of an air pump, and the normal pressure port 160 may be provided in the air.
The flow cytometer provided by the present invention is illustrated by the following four examples.
The present invention proposes a first embodiment, which is described below with reference to fig. 1.
The flow cytometer also includes an injection assembly 180 for controlling the flow of the fluid to be detected into the flow chamber 120, a fourth conduit 240, and a fifth conduit 250. Two ends of the fourth pipeline 240 are respectively communicated with the injection assembly 180 and the reaction tank 110, and two ends of the fifth pipeline 250 are respectively communicated with the flow chamber 120 and the fourth pipeline 240.
Further, the injection assembly 180 includes a first syringe 181 and a second syringe 182, and the first syringe 181 is communicated with the second syringe 182 through a sixth pipeline 260.
Further, the flow cytometer also includes a seventh conduit 270, the seventh conduit 270 being in communication with the sixth conduit 260 and the flow cell 120. The seventh pipeline 270 is used for inputting a diluent to facilitate analysis of the liquid to be detected.
To further refine the structure, a first valve 410 for controlling the on-off of the first pipeline 210 is provided on the first pipeline 210. The third pipeline 230 is provided with a second valve 420 for controlling the on-off of the third pipeline 230. The fourth pipeline 240 is provided with a third valve 430 for controlling the on-off of the fourth pipeline 240, and the third valve 430 is arranged between the reaction tank 110 and the fifth pipeline 250. The sixth pipeline 260 is provided with a fourth valve 440 for controlling the on-off of the sixth pipeline 260. The seventh pipeline 270 is provided with a fifth valve 450 for controlling the on-off of the seventh pipeline 270.
Specifically, when the liquid to be detected is inputted, the third valve 430 and the fourth valve 440 are opened, and the first syringe 181 operates to suck the liquid to be detected in the reaction tank 110 into the third pipe 230. The third valve 430 is then closed and the second syringe 182 is operated to push the liquid to be tested into the flow chamber 120 from the fifth line 250. At the same time, the fifth valve 450 is opened and the first syringe 181 is operated to push the diluent into the flow chamber 120 so that the diluent mixes with the liquid to be tested to form a sheath flow. After the analysis is completed, the flow cell 120 discharges the waste liquid.
When waste liquid is input into the liquid tank 130, the first valve 410 is opened, the second valve 420 is closed, and the three-way valve 140 is communicated with the normal pressure port 160. Next, the waste liquid enters the liquid tank 130 through the first pipe 210, and the liquid tank 130 temporarily stores the waste liquid.
When the liquid tank 130 outputs waste liquid, the first valve 410 is closed, the second valve 420 is opened, and the three-way valve 140 is communicated with the positive pressure port 150. At this time, the waste liquid enters the waste liquid interface 170 from the third pipe 230 under the pressure to discharge the waste liquid.
In addition, after the analysis is completed, the line in the flow cytometer is cleaned. Specifically, the injection assembly 180 operates to draw in the diluent, and then the injection assembly 180 passes the diluent into the fourth line 240 to clean the fourth line 240, and finally is discharged from the reaction cell 110.
Based on the first embodiment, the present invention proposes a second embodiment, which will be explained below with reference to fig. 2.
In the present embodiment, the flow cytometer further includes an eighth pipeline 280, wherein the eighth pipeline 280 communicates with the third pipeline 230 and the fourth pipeline 240, and an end communicating with the fourth pipeline 240 is disposed between the injection assembly 180 and the fifth pipeline 250.
To achieve better effect, a sixth valve 460 is also provided on the eighth conduit 280.
To conserve diluent, waste liquid is used in this embodiment to purge the fourth line 240. It will be appreciated that the waste liquid is composed of approximately 100ul of reaction solution with a cell particle content of about 1:150 and 7000ul of clean diluent, so that the cell particle number in the waste liquid is very low, and the main component is diluent, so that the waste liquid can be used to clean the fourth pipeline 240, and then a small amount of diluent can be used to clean the fourth pipeline 240 again.
Specifically, the three-way valve 140 communicates with the positive pressure port 150, the first valve 410 is closed, the second valve 420 is closed, the fourth valve 440 is closed, the third valve 430 is open, and the sixth valve 460 is open. The waste liquid enters the fourth pipeline 240 from the eighth pipeline 280 under the pressure of the positive pressure port 150, so as to clean the fourth pipeline 240, and then is discharged from the reaction tank 110.
By adopting the structure, the waste liquid utilization is realized, meanwhile, the use of the diluent is also saved, and the use cost is reduced. On the one hand, as the liquid is continuously pushed by using positive pressure, the liquid can be always cleaned, the cleaning amount is large, the cleaning time is short, the action similar to a syringe is not needed, and repeated suction-discharge-suction-discharge is needed; on the other hand, because the waste liquid with small cell particle number is cleaned by using the positive pressure pushing, only the final cleaning is needed to be performed by using a large injector to provide partial diluent for flushing, so that the diluent can be saved, and the utilization rate of the reagent can be improved.
Based on the second embodiment, the present invention proposes a third embodiment, which will be explained below with reference to fig. 3.
The flow cytometer also includes a swab 510, a ninth conduit 290, and a tenth conduit 300; the swab 510 includes a wiping part 512 for cleaning and a sampling needle 511 fixedly connected with the wiping part 512, the sampling needle 511 is movable along an extending direction thereof to enable the wiping part 512 to complete cleaning, two ends of the ninth pipeline 290 are respectively communicated with the third pipeline 230 and the wiping part 512, one end of the tenth pipeline 300 is communicated with the wiping part 512, and the other end of the tenth pipeline 300 is communicated with a waste liquid pump for communicating treatment waste liquid.
Further, a seventh valve 470 is provided on the ninth conduit 290.
It will be appreciated that by this method, the internal cleaned waste liquid of the flow cytometer may be used for other cleaning components as well, such as FIG. 3, where the collected waste liquid from the cleaning flow cell is used to clean the swab 510. When the flow chamber needs to consume sheath liquid for cleaning, not only the cell re-detection is carried out, but also the flow of cleaning the instrument by switching on and switching off, and the main purpose of the diluent cleaning the flow chamber at the moment is to clean and remove bubbles in the flow chamber and a pipeline thereof, so that the flow chamber is completely full of liquid, and the waste liquid collected at the moment is free of cell particles, can be completely used as a cleaning agent of other units, and does not produce carrying pollution.
Specifically, the three-way valve 140 communicates with the positive pressure port 150, the seventh valve 470 is open, and the remaining valves are closed. Under the action of pressure, the waste liquid enters the ninth pipeline 290 from the third pipeline 230, then enters the wiping part 512, and then the external cleaning is realized by moving the sampling needle 511. And the waste liquid finally flows out of the tenth pipe 300.
Referring to fig. 4, the present invention also provides a fourth embodiment, and the technical solution provided by the embodiment can be used to unblock the jewel hole of the impedance channel.
The flow cytometer further includes an eleventh pipeline 310 and a WBC tank 520, and both ends of the eleventh pipeline 310 are respectively connected to the third pipeline 230 and the WBC tank 520.
Further, an eighth valve 480 is disposed on the eleventh pipeline 310, and a ninth valve 490 is additionally disposed on the third pipeline 230.
It should be noted that WBC pool 520 is a white blood cell pool.
In addition, when a hole blockage occurs in the WBC pool 520, the eighth valve 480 is opened, the ninth valve 490 is opened, the rest valves are closed, and the three-way valve 140 is communicated with the positive pressure port 150. At this time, the left side of the third pipe 230 is in a closed state, and the pressure in the pipe is the pressure provided by the positive pressure port 150, that is, the pressure directly acts on the small hole of the WBC pool 520 relative to the positive pressure port 150, and the plugged WBC jewel hole is backflushed, and generally, the pressure can be built to 50kpa. Most of the slightly blocked jewel holes can be solved by the method.
In this embodiment, another method of backflushing a WBC pool plug is also provided. The first injector 181 is advanced back, the first valve 410, fourth valve 440, fifth valve 450, eighth valve 480 are opened, the remaining valves are closed, and the flow direction of the liquid acts on the jewel orifice of the blocked WBC cell along the fourth line 240, eighth line 280, eleventh line 310. Because the flow chamber is constructed as an elongated orifice, the fluid flow through such orifice is required to overcome a 120kpa pressure differential, and the eighth conduit 280 and the seventh conduit 270 are in communication, the pressures are identical, i.e., the pressure of 120kpa generated by the flow of the first syringe 181 through the flow chamber is used to recoil the gemstone orifice, and the flow chamber is then used as a 120kpa overflow valve, utilizing the pressure differential function within the instrument itself to effect the recoil and cleaning of the gemstone orifice.
The foregoing disclosure is illustrative of the present invention and is not to be construed as limiting the scope of the invention, which is defined by the appended claims.
Claims (7)
1. The flow cytometry analyzer is characterized by comprising a reaction tank for reacting a liquid to be analyzed, a flow chamber for analyzing the liquid to be analyzed after the reaction in the reaction tank, a liquid tank for temporarily storing waste liquid discharged from the flow chamber, a first pipeline, a second pipeline, a third pipeline and a three-way valve;
two ends of the first pipeline are respectively communicated with the flow chamber and the liquid tank, two ends of the second pipeline are respectively communicated with one of the liquid tank and the three-way valve, the other two ports in the three-way valve are respectively used for communicating a positive pressure port and a normal pressure port, one end of the third pipeline is communicated with the liquid tank, and the other end of the third pipeline is used for communicating a waste liquid interface;
when the liquid tank receives the waste liquid, the three-way valve is communicated with the normal pressure port, so that the waste liquid stably flows into the liquid tank; when the waste liquid in the liquid tank is discharged, the three-way valve is communicated with the positive pressure port, so that the waste liquid enters the waste liquid interface through the third pipeline;
the flow cytometer also comprises an injection assembly, a fourth pipeline and a fifth pipeline; two ends of the fourth pipeline are respectively communicated with the injection assembly and the reaction tank, and two ends of the fifth pipeline are respectively communicated with the flow chamber and the fourth pipeline;
a third valve for controlling the on-off of the fourth pipeline is arranged on the fourth pipeline, and the third valve is arranged between the reaction tank and the fifth pipeline;
the flow cytometry further comprises an eighth pipeline, wherein two ends of the eighth pipeline are respectively communicated with the third pipeline and the fourth pipeline, and one end of the eighth pipeline, which is communicated with the fourth pipeline, is positioned between the injection assembly and the fifth pipeline.
2. The flow cytometer of claim 1 wherein the first conduit is provided with a first valve for controlling the on-off of the first conduit and the third conduit is provided with a second valve for controlling the on-off of the third conduit.
3. The flow cytometer of claim 1 wherein the injection assembly comprises a first syringe, a second syringe, and a sixth conduit; the two ends of the sixth pipeline are respectively communicated with the first injector and the second injector, and the second injector is communicated with the fourth pipeline.
4. The flow cytometer of claim 3 wherein the sixth conduit is provided with a fourth valve for controlling the on-off of the sixth conduit.
5. The flow cytometer of claim 4 further comprising a seventh conduit that communicates with the sixth conduit and the flow chamber such that the seventh conduit is capable of providing a diluent to the flow chamber.
6. The flow cytometer of any of claims 1-5, further comprising a swab, a ninth conduit, and a tenth conduit; the swab comprises a wiping part for cleaning and a sampling needle fixedly connected with the wiping part, the sampling needle can move along the extending direction of the sampling needle so that the wiping part can finish cleaning, two ends of a ninth pipeline are respectively communicated with the third pipeline and the wiping part, one end of the tenth pipeline is communicated with the wiping part, and the other end of the tenth pipeline is communicated with a waste liquid pump for communicating and treating waste liquid.
7. The flow cytometer of any of claims 1-5 further comprising an eleventh conduit and a WBC reservoir, wherein the ends of the eleventh conduit are in communication with the third conduit and the WBC reservoir, respectively.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110332240.1A CN113125330B (en) | 2021-03-29 | 2021-03-29 | Flow cytometry analyzer |
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| CN202110332240.1A CN113125330B (en) | 2021-03-29 | 2021-03-29 | Flow cytometry analyzer |
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| CN113125330B true CN113125330B (en) | 2023-09-19 |
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| CN116087549A (en) * | 2023-04-12 | 2023-05-09 | 深圳市帝迈生物技术有限公司 | Control method of sample analyzer and sample analyzer |
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