CN113884427A - Sheath flow impedance particle analyzer and sample measuring method thereof - Google Patents
Sheath flow impedance particle analyzer and sample measuring method thereof Download PDFInfo
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- CN113884427A CN113884427A CN202010725172.0A CN202010725172A CN113884427A CN 113884427 A CN113884427 A CN 113884427A CN 202010725172 A CN202010725172 A CN 202010725172A CN 113884427 A CN113884427 A CN 113884427A
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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
- G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
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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—Optical investigation techniques, e.g. flow cytometry
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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—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
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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—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1486—Counting the particles
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Abstract
The invention provides a sheath flow impedance particle analyzer and a sample measuring method thereof. The invention can simplify the structure of the sheath flow impedance particle analyzer, and realize the parallel execution of the emptying operation of the back sheath waste liquid pool and the measurement operation of the sheath flow impedance counting pool, thereby achieving the dual purposes of improving the sample measurement speed and reducing the cost and the volume of the analyzer.
Description
The present application claims priority from the chinese patent application entitled "sheath flow impedance method particle analyzer and liquid level initialization method thereof" filed by the chinese patent office on 03/07/2020, application No. 202010635511.6, which is incorporated herein by reference in its entirety.
Technical Field
The invention relates to the technical field of particle analysis equipment, in particular to a sheath flow impedance particle analyzer and a sample measuring method thereof.
Background
The existing particle analyzer by the sheath flow impedance method generally comprises a premixing pool, a sample needle, a sheath flow impedance counting pool, a back sheath waste liquid pool, a waste liquid pool and the like.
The measurement process of the sheath flow impedance method particle analyzer mainly comprises the following steps: cleaning a premixing pool and discharging waste to a waste liquid pool, cleaning a sheath flow impedance counting pool and discharging waste to the waste liquid pool and a rear sheath waste liquid pool, adding a sample to the premixing pool and uniformly mixing, preparing the sample by a sample needle, forming front and rear sheath liquids, pushing the sample by the sample needle to the sheath flow impedance counting pool for measurement, and discharging the liquid to the rear sheath waste liquid pool, the rear sheath waste liquid pool and the like in the measurement process.
Disclosure of Invention
Referring to fig. 1, the sheath flow impedance counting cell in the prior art includes a front cell, a rear cell, and a partition plate between the front cell and the rear cell, the partition plate being provided with a jewel hole. The back sheath channel connected to the back cell of the sheath flow impedance counting cell will typically include two isolation cells: a back sheath isolation tank for injecting sheath liquid into the back sheath liquid port, and a back sheath waste liquid tank for collecting waste liquid discharged from the back tank waste liquid port.
In the process of sample measurement operation or cleaning of the sheath flow impedance counting cell, a certain amount of waste liquid is collected by the back sheath waste liquid cell, and in order to avoid influencing subsequent sample measurement operation, the back sheath waste liquid cell needs to be emptied.
Referring to fig. 1, the operation process may include: closing the valve V10 to isolate the back sheath waste liquid pool and the sheath flow impedance counting pool, opening a control valve (not shown in figure 1) between the waste liquid pool and the negative pressure source, opening the valve V7 and the valve V11, conducting the back sheath waste liquid pool and the negative pressure source in the waste liquid pool, emptying the back sheath waste liquid pool after a certain time, then closing the control valve between the waste liquid pool and the negative pressure source, closing the valve V7 and the valve V11, and opening the valve V10.
The valve V10 must be closed before the sheath waste liquid pool is emptied by using negative pressure to ensure that the sheath flow impedance counting pool is not affected by the negative pressure (the negative pressure generated in the sheath flow impedance counting pool affects the sample measurement), and the valve V7 and the valve V11 are closed after the sheath waste liquid pool is emptied to restore the sheath waste liquid pool to the normal pressure state, the valve V10 is opened so that the sheath flow impedance counting pool can measure in the normal pressure state, and the waste liquid in the sheath flow impedance counting pool can flow into the sheath waste liquid pool through the valve V10.
In addition, in the process of discharging liquid from the rear sheath waste liquid pool by using the negative pressure source, the valve V11 must be opened to keep a certain air suction flow rate so as to ensure that the waste liquid in the rear sheath waste liquid pool is pumped away, otherwise, after the negative pressure liquid is discharged for a period of time, the pressure in the rear sheath waste liquid pool is balanced (no negative pressure), and the rear sheath waste liquid pool becomes a closed space and cannot be discharged continuously.
Since the valve V10 is closed and the sheath flow impedance counting cell cannot perform measurement in the negative pressure environment, before the sheath waste liquid cell is emptied by the negative pressure source, the measurement operation in the sheath flow impedance counting cell needs to be stopped, that is, the emptying operation needs to be performed independently outside the measurement operation, and the measurement operation and the emptying operation cannot be performed in parallel, which results in low efficiency of sample measurement. Moreover, the sheath flow impedance method particle analyzer requires the valve V10 and the valve V11 to be able to perform the evacuation operation, which results in an increase in the instrument cost and volume of the sheath flow impedance method particle analyzer.
In view of this, the present invention provides a sheath flow impedance particle analyzer and a sample measuring method thereof, which can delete the valve V10 and the valve V11 on the basis of the existing sheath flow impedance particle analyzer, and implement the parallel execution of the emptying operation of the back sheath waste liquid pool and the measuring operation of the sheath flow impedance counting pool, so as to increase the sample measuring speed and reduce the instrument cost and volume.
In order to solve the above technical problems, the present invention provides the following technical features:
in order to achieve the above object, the present invention provides the following technical features:
a sheath flow impedance particle analyzer comprising:
the system comprises a premixing pool, a sample needle, a sheath flow impedance counting pool, a sheath liquid pool, a rear sheath isolation pool, a rear sheath waste liquid pool and a waste liquid pool, wherein the premixing pool is connected to an input port of the sample needle;
the sheath flow impedance counting cell comprises a front cell and a back cell, the first output port of the sample needle is connected to the front cell, the front cell comprises a front sheath fluid inlet and a front sheath fluid outlet, the front sheath fluid inlet is connected to the sheath fluid cell through a front cell sheath fluid channel to provide sheath fluid into the front cell through the front cell sheath fluid channel and the front sheath fluid inlet, the front sheath fluid outlet is connected to the waste fluid cell through a front cell cleaning channel, and the front cell cleaning channel is provided with a first control valve;
the rear pool comprises a rear pool sheath liquid port and a rear pool waste liquid port, the rear pool sheath liquid port is connected to the rear sheath isolation pool, the rear sheath isolation pool is connected to the sheath liquid pool through a rear pool sheath liquid channel, and sheath liquid is provided into the rear pool through the rear pool sheath liquid channel, the rear sheath isolation pool and the rear pool sheath liquid port;
the rear sheath waste liquid tank is connected with the rear sheath waste liquid tank through a rear sheath liquid discharge channel, and the rear sheath liquid discharge channel is provided with a second control valve;
in the sample measuring process of the sheath flow impedance counting cell, the rear sheath isolation cell, the rear sheath waste liquid cell and the waste liquid cell are in a closed environment and normal pressure state.
Optionally, the waste liquid pool is connected with a negative pressure source, and a third control valve is arranged between the waste liquid pool and the negative pressure source, so that the third control valve is closed to switch the waste liquid pool to a normal pressure state, and the third control valve is opened to switch the waste liquid pool to a negative pressure state.
Optionally, the waste liquid pool includes a normal pressure waste liquid pool and a negative pressure waste liquid pool, and the second control valve is a three-way valve, one end of the three-way valve is connected with the back sheath waste liquid pool, and the other two ends are respectively connected with the normal pressure waste liquid pool and the negative pressure waste liquid pool.
Optionally, the method further includes:
and the fourth control valve is arranged at the upper part of the rear sheath isolation pool and is communicated with air, so that the fourth control valve is opened to communicate the air with the rear sheath isolation pool, and the fourth control valve is closed to isolate the air.
Optionally, the method further includes:
the rear sheath isolation pool is connected to the sheath fluid pool through a rear pool cleaning channel;
the sheath fluid bath is connected to a positive pressure source.
Optionally, the method further includes:
a sample preparation power source connected to the second output port of the sample needle via a sample preparation channel; the sample preparation channel is provided with a fifth control valve.
Optionally, the method further includes:
the output port of the sample pushing injector is connected to the first input port of the sample needle, the input port of the sample pushing injector is connected to the sheath fluid pool through a sample pushing channel, and the sample pushing channel is provided with a sixth control valve.
A sample measurement method for use with the sheath flow impedance particle analyzer, the method comprising:
adjusting the waste liquid pool to be in a normal pressure state;
opening a second control valve between the rear sheath waste liquid pool and the waste liquid pool to control the rear sheath isolation pool, the rear sheath waste liquid pool and the waste liquid pool to be in a closed environment and normal pressure state;
and controlling the sheath flow impedance counting cell to perform measurement operation on the sample and continuously perform liquid drainage operation on the rear sheath waste liquid pool so as to continuously accumulate waste liquid in the rear sheath waste liquid pool, and automatically performing liquid drainage operation on the waste liquid pool under the condition that the pressure in the rear sheath waste liquid pool is greater than liquid drainage resistance in a closed environment.
Optionally, after the sheath flow impedance particle analyzer is started, performing a liquid level initialization operation on the back sheath isolation pool;
wherein the performing a level initialization operation on the back sheath isolation tank comprises:
emptying the diluent in the sheath fluid pool;
evacuating liquid from the back sheath isolation tank and the back sheath waste liquid tank;
filling the sheath liquid pool and filling a channel connected with the sheath flow impedance counting pool;
setting the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool to preset negative pressure;
adding a preset amount of diluent into the rear sheath isolation pool through a rear sheath cleaning channel;
setting the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool to be in a normal pressure state;
and after the liquid level in the back sheath isolation pool is stable, finishing the liquid level initialization of the back sheath isolation pool.
Optionally, the sheath flow impedance particle analyzer further includes a fourth control valve disposed at an upper portion of the rear sheath isolation pool and connected to air;
the sample measuring method further comprises the step of performing liquid level initialization operation on the rear sheath isolation pool after the sheath flow impedance particle analyzer is started;
wherein the performing a level initialization operation on the back sheath isolation tank comprises:
arranging the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool to be in a negative pressure state, and emptying liquid in the rear sheath isolation chamber and the rear sheath waste liquid pool;
setting the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool as preset negative pressure;
adding a preset amount of diluent into the rear sheath isolation pool through a rear sheath cleaning channel;
setting the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool to be in a normal pressure state;
and after the liquid level in the back sheath isolation pool is stable, finishing the liquid level initialization of the back sheath isolation pool.
The invention provides a sheath flow impedance particle analyzer, which is characterized in that a valve V10 and a valve V11 are deleted on the basis of the original sheath flow impedance particle analyzer, so that the structure of the sheath flow impedance particle analyzer is simplified, and the purposes of reducing the cost and the volume of the analyzer are achieved.
On the basis that the valve V10 and the valve V11 are deleted by the sheath flow impedance particle analyzer, in the sample measurement process of the sheath flow impedance counting cell, the back sheath isolation cell, the back sheath waste liquid cell and the waste liquid cell are in a sealed environment and in a normal pressure state, the sheath flow impedance counting cell can normally carry out measurement in the normal pressure state, and in addition, after the valve V10 is deleted, the liquid discharge operation can be normally carried out on the back sheath waste liquid cell in the measurement process.
In the sample measurement process of the sheath flow impedance counting cell, the back sheath isolation cell, the back sheath waste liquid cell and the waste liquid cell are in a closed environment, so that waste liquid is continuously accumulated in the back sheath waste liquid cell in the process of continuously performing liquid drainage operation on the back sheath waste liquid cell by the sheath flow impedance counting cell, and the purpose of automatically performing liquid drainage operation on the waste liquid cell is realized under the condition that the pressure in the back sheath waste liquid cell is greater than liquid drainage resistance in the closed environment.
The invention can simplify the structure of the sheath flow impedance particle analyzer, and realize the parallel execution of the emptying operation of the back sheath waste liquid pool and the measurement operation of the sheath flow impedance counting pool, thereby achieving the dual purposes of improving the sample measurement speed and reducing the cost and the volume of the analyzer.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic diagram of a sheath flow impedance particle analyzer;
FIG. 2 is a schematic structural diagram of a first embodiment of a sheath flow impedance particle analyzer provided by the present invention;
FIG. 3 is a schematic structural diagram of a second embodiment of a sheath flow impedance particle analyzer provided by the present invention;
FIG. 4 is a schematic structural diagram of a third embodiment of a sheath flow impedance particle analyzer provided by the present invention;
FIG. 5 is a flow chart of a sample measurement method provided by the present invention;
FIG. 6 is a flow chart of a first embodiment of liquid level initialization provided by the present invention;
FIGS. 7a-7c are schematic structural views of a fourth embodiment to a sixth embodiment of a sheath flow impedance particle analyzer according to the present invention;
FIG. 8 is a flow chart of a second embodiment of liquid level initialization provided by the present invention.
Detailed Description
The terms in the detailed description are to be construed as:
anterior pool sheath fluid channel: a pipeline between the sheath liquid pool 7 and the front sheath liquid inlet 43, and a valve V2 is arranged on a sheath liquid channel of the front pool.
A forebay cleaning channel: a pipeline between the front sheath liquid outlet 46 and the waste liquid pool 9, and a valve V8 is arranged on the front pool sheath liquid channel.
Back pool sheath fluid channel: a pipeline between the sheath liquid pool 7 and the back sheath isolation pool 6, and a valve V4 is arranged on a sheath liquid channel of the back pool
Rear sheath drainage channel: and a valve V3 is arranged on a sheath liquid channel of the rear pool of the other pipeline between the sheath liquid pool 7 and the rear sheath isolation pool 6.
And (3) cleaning a channel in a rear pool: and a valve V7 is arranged on a pipeline between the rear sheath waste liquid pool 5 and the waste liquid pool 9 and a cleaning channel of the rear pool.
Sample preparation channel: the sample preparation channel, which is a conduit between the sample needle 3 and the sample preparation power source 11, is provided with a valve V6.
Sample liquid pushing channel: a pipeline between the sheath fluid pool 7 and the sample pushing injector 2, and a valve V5 is arranged on a sample pushing channel.
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without inventive effort based on the embodiments of the present invention, are within the scope of the present invention.
References herein to "connected to" include direct connections and connections made indirectly through other components, such as control valves.
Referring to fig. 2, fig. 2 is a schematic diagram of a first embodiment of a sheath flow impedance particle analyzer according to the present invention.
The embodiment provides a particle analyzer by a sheath flow impedance method, which comprises a premixing pool 1, a sample needle 3, a sheath flow impedance counting pool 4, a sheath liquid pool 7, a rear sheath isolation pool 6, a rear sheath waste liquid pool 5 and a waste liquid pool 9. The premix cell 1 is connected to the input port 31 of the sample needle 3, and a valve V1 is provided between the premix cell 1 and the input port 31 of the sample needle 3.
The sheath flow impedance counting cell 4 includes a front cell 41 and a rear cell 42, and the first output port 33 of the sample needle 3 is connected to the front cell 41; the anterior chamber 41 includes an anterior chamber fluid inlet 43 and an anterior chamber fluid outlet 46, the anterior chamber fluid inlet 43 being connected to the sheath fluid chamber 7 via an anterior chamber sheath fluid channel for providing sheath fluid into the anterior chamber 41 through the anterior chamber sheath fluid channel and the anterior sheath fluid inlet 43. The front pool sheath liquid channel is provided with a valve V2.
The sheath fluid outlet 46 is connected to the waste fluid tank 9 via a tank cleaning passage, and the tank cleaning passage is provided with a first control valve (valve V8).
The rear sheath waste liquid port 45 is connected to the rear sheath waste liquid tank 5, the rear sheath waste liquid tank 5 is connected to the waste liquid tank 9 through a rear sheath liquid discharge channel, and the rear sheath liquid discharge channel is provided with a second control valve (valve V7). According to the actual use condition, the waste liquid pool can be adjusted to be in a normal pressure state or a negative pressure state.
Further, the particle analyzer by sheath flow impedance method also comprises the following components:
a partition plate 47 is arranged between the front tank 41 and the rear tank 42 of the sheath flow impedance counting tank 4, and a gem hole is arranged on the partition plate 47. The first output port 33 of the sample needle 3 is located opposite the keyhole.
A sample preparation power source 11, and the sample preparation power source 11 is connected to the second output port 32 of the sample needle 3 via a sample preparation channel; the sample preparation channel is provided with a fifth control valve (valve V6). Specifically, the sample preparation power source 11 may be a fixed displacement pump, a syringe, a liquid pump, or a peristaltic pump.
The rear sheath isolation tank 6 is connected to the sheath fluid tank 7 through a rear tank cleaning passage, and the rear tank cleaning passage is provided with a valve V3. The sheath fluid reservoir 7 is connected to a positive pressure source 8.
A sample push syringe 2, and an output port of the sample push syringe 2 is connected to the input port 31 of the sample needle 3, the input port of the sample push syringe 2 is connected to the sheath fluid pool 7 via a sample push channel, and the sample push channel is provided with a sixth control valve (valve V5).
According to the technical characteristics, the present embodiment has at least the following beneficial effects:
the present embodiment provides a sheath flow impedance particle analyzer, which deletes the valve V10 and the valve V11 on the basis of the original sheath flow impedance particle analyzer, thereby simplifying the structure of the sheath flow impedance particle analyzer and achieving the purpose of reducing the cost and volume of the analyzer.
Referring to fig. 3, fig. 3 is a schematic diagram of a particle analyzer using sheath flow impedance method according to a second embodiment of the present invention.
On the basis of fig. 2, a specific implementation of the waste liquid tank 9 is provided.
Waste liquid pond 9 is connected in the negative pressure source, just, waste liquid pond with be provided with third control valve (valve V10) between the negative pressure source, so that close the third control valve can switch waste liquid pond is normal pressure state, opens the third control valve can switch waste liquid pond is negative pressure state.
When it is necessary to control the waste liquid tank 9 to be in a normal pressure state, a controller (not shown) of the sheath flow impedance method particle analyzer closes the third control valve, so that the waste liquid tank becomes in a normal pressure state. When the waste liquid pool 9 is required to be controlled to be in a negative pressure state, the controller of the sheath flow impedance method particle analyzer opens the third control valve, so that the waste liquid pool is changed to be in the negative pressure state.
Referring to fig. 4, fig. 4 is a schematic diagram of a third embodiment of a sheath flow impedance particle analyzer according to the present invention.
The waste liquid pool 9 comprises a normal-pressure waste liquid pool 91 and a negative-pressure waste liquid pool 92, the second control valve is a three-way valve, one end of the three-way valve is connected with the back sheath waste liquid pool 5, and the other two ends of the three-way valve are respectively connected with the normal-pressure waste liquid pool 91 and the negative-pressure waste liquid pool 92.
When the waste liquid pool 9 is required to be controlled to be in a normal pressure state, the controller of the sheath flow impedance method particle analyzer controls the three-way valve to be communicated with the normal pressure waste liquid pool 91 to be in the normal pressure state; when the waste liquid pool 9 is required to be controlled to be in a negative pressure state, the controller of the sheath flow impedance particle analyzer controls the three-way valve to be communicated with the negative pressure waste liquid pool 92 to be in the negative pressure state.
According to the technical characteristics, the present embodiment has at least the following beneficial effects:
the second embodiment and the third embodiment respectively provide two implementation manners of the waste liquid pool, and both the two implementation manners can adjust the waste liquid pool to be in a negative pressure state or a normal pressure state.
Referring to fig. 5, the present invention provides a sample measurement method applied to any one of the sheath flow impedance particle analyzers of fig. 2-4. The sample measuring method comprises the following steps:
steps S501 to S503 are preparatory steps for performing sample measurement, and step S504 is a real step for performing sample measurement.
Step S501: and initializing the liquid level of the back sheath isolation pool by the sheath flow impedance method particle analyzer.
When the particle analyzer is used for the first time or after maintenance, the liquid level initialization operation needs to be performed on the back sheath isolation pool 6 so as to enable the back sheath isolation pool 6 to have a proper liquid level. Air is arranged above the liquid level of the rear sheath isolation pool 6, and an electric isolation effect can be achieved.
Step S502: a cleaning operation is performed on the sheath flow impedance counting cell.
Washing anterior pool 41 of sheath flow impedance counting pool 4:
a controller (not shown) controls opening valve V2, valve V8 and activates positive pressure source 8. Sheath fluid in sheath fluid reservoir 7 is supplied to anterior reservoir 41 through the anterior reservoir sheath fluid channel and anterior sheath fluid inlet 43 under positive pressure from positive pressure source 8. The sheath fluid is drained through the front sheath fluid outlet 46 to the waste reservoir 9 during the cleaning process.
Cleaning of the rear tank 42 of the sheath flow impedance counting tank 4:
a controller (not shown) controls the open valve V3 and activates the positive pressure source 8. Under the positive pressure of the positive pressure source 8, the sheath fluid in the sheath fluid tank 7 is conveyed into the rear tank 42 through the rear tank cleaning channel, the rear sheath isolation tank 6 and the rear tank sheath fluid port 44, and the cleaning operation is performed on the rear tank 42. The sheath liquid will be discharged to the rear sheath waste liquid tank 5 through the rear tank waste liquid port 45 during the cleaning process.
And (3) performing emptying operation on the rear sheath waste liquid pool 5 in the cleaning process:
the back sheath waste liquid tank 5 accumulates waste liquid in the cleaning process of the cleaning sheath flow impedance counting tank 4, so that the back sheath waste liquid tank 5 needs to be emptied in the cleaning process. The controller controls the waste liquid pool 9 to be in a negative pressure state, opens the valve V7, and empties the waste liquid in the back sheath waste liquid pool 5 in the negative pressure state.
In the second embodiment shown in fig. 3, the third control valve V10 between the waste liquid pool 9 and the negative pressure source may be opened to switch the waste liquid pool 9 to the negative pressure state. In a third embodiment shown in fig. 4, the three-way valve may be switched to a negative pressure waste reservoir to a negative pressure state.
Since the measurement operation is started only after the cleaning operation of the sheath flow impedance counting cell is completed, the back sheath waste liquid pool 5 can be emptied by directly using the negative pressure during the cleaning operation of the sheath flow impedance counting cell.
Step S503: a sample to be tested is prepared.
The premixed tank 1 is provided with the uniformly mixed sample. The controller opens valve V1 and valve V6, draws sample from premix cell 1 into sample needle 3 under the action of sample preparation power source 11, and closes valve V1 and valve V6.
The controller opens valve V5, and the sample push syringe 2 draws sheath fluid from the sheath fluid reservoir 7 and then pushes the sheath fluid to the sample needle 3 to push the sample in the sample needle 3 into the anterior reservoir 41 of the sheath fluid impedance counter 4.
In the sample measurement process of the sheath flow impedance counting cell, the rear sheath isolation cell 6, the rear sheath waste liquid cell 5 and the waste liquid cell 9 are in a closed environment and at normal pressure, and specifically include steps S504 to S506.
Step S504: the controller adjusts the waste liquid tank to be in a normal pressure state.
In order to avoid the negative pressure from influencing the measurement operation and causing inaccurate measurement results, the waste liquid tank 9 is adjusted to be in a normal pressure state before measurement.
In the second embodiment shown in fig. 3, the third control valve V10 between the waste liquid tank 9 and the negative pressure source may be closed to switch the waste liquid tank 9 to the normal pressure state. In a third embodiment shown in fig. 4, the three-way valve can be switched to the atmospheric waste liquid tank to change to the atmospheric state.
Step S505: and opening a second control valve between the rear sheath waste liquid pool and the waste liquid pool to control the rear sheath isolation pool, the rear sheath waste liquid pool and the waste liquid pool to be in a closed environment and normal pressure state.
The second control valve (valve V7) between the sheath waste reservoir and the waste reservoir was opened, which in the second instance shown in fig. 3 required opening of valve V7. In the third embodiment shown in fig. 4, the valve V7 is already opened when the three-way valve is switched to the atmospheric waste liquid pool in step S504.
Since the rear sheath separation tank 6, the rear sheath waste liquid tank 5, and the waste liquid tank 9 are in a communicated state after the valve V7 is opened, the rear sheath separation tank 6, the rear sheath waste liquid tank 5, and the waste liquid tank 9 can be kept in a normal pressure state even when the waste liquid tank is in a normal pressure state.
After the valve V10 and the valve V11 are deleted in the structure of the particle analyzer adopting the sheath flow impedance method, the rear sheath isolation pool 6 and the rear sheath waste liquid pool 5 are not communicated with air, so that the rear sheath isolation pool 6, the rear sheath waste liquid pool 5 and the waste liquid pool 9 are in a closed environment.
Step S506: the sample measurement operation and the evacuation operation are started. Namely: the sheath flow impedance counting cell performs measurement operation on a sample and continuously performs liquid drainage operation on the rear sheath waste liquid pool so as to enable the rear sheath waste liquid pool to continuously accumulate waste liquid, and liquid drainage operation is automatically performed on the waste liquid pool under the condition that the pressure in the rear sheath waste liquid pool is greater than liquid drainage resistance in a closed environment.
During the measuring operation, the positive pressure source 8, the valve V2 and the valve V4 are opened.
Under the action of the positive pressure source 8, the sheath fluid reservoir 7 inputs the sheath fluid into the anterior reservoir 41 through the valve V2 and the anterior reservoir sheath fluid inlet 43, and forms a sheath fluid flow in the anterior reservoir 41. The sheath fluid surrounds the sample introduced through the sample needle 3 through the jewel aperture so that particles in the sample line up through the jewel aperture for accurate measurement of the sample.
During the measurement operation, the sheath fluid reservoir 7 feeds sheath fluid into the back reservoir 42 through the valve V4, the back sheath isolation reservoir 6, and the back reservoir sheath fluid port 44 under the action of the positive pressure source 8, and forms a sheath fluid flow in the back reservoir, which wraps around the sample flow coming out of the jewel aperture (the sheath fluid flow wraps around the sample fed through the sample needle 3), and flows into the back sheath waste reservoir 5.
Because the back sheath isolation pool 6, the back sheath waste liquid pool 5 and the waste liquid pool 9 are in a closed environment, and in the measuring process, the sheath flow impedance counting pool 4 continuously executes liquid drainage operation to the back sheath waste liquid pool 5, waste liquid is continuously accumulated in the back sheath waste liquid pool 5, and when the pressure in the back sheath waste liquid pool 5 in the closed environment is greater than the liquid drainage resistance, the liquid drainage operation is automatically executed to the waste liquid pool 9.
According to the technical characteristics, the present embodiment has at least the following beneficial effects:
in the embodiment, the back sheath waste liquid pool 5 is not evacuated by using negative pressure, but the back sheath waste liquid pool 5 is set to be in a normal pressure state, and the sample measurement operation of the sheath flow impedance counting pool is not influenced in the normal pressure state, so that the purpose of parallel execution of evacuation operation and measurement operation can be realized.
In this embodiment, no additional power source is used to empty the back sheath waste liquid pool, but the waste liquid is continuously discharged by means of the measurement process of the sheath flow impedance counting cell 4 to increase the pressure in the back sheath waste liquid pool 5, and when the pressure in the back sheath waste liquid pool 5 in the closed environment is greater than the liquid discharge resistance, the back sheath waste liquid pool 5 can automatically discharge liquid to the waste liquid pool 9.
According to the technical characteristics, the present embodiment has at least the following beneficial effects:
on the basis of any one of the sheath flow impedance particle analyzers shown in fig. 2-4, in the embodiment, the evacuation operation of the rear sheath waste liquid pool and the measurement operation of the sheath flow impedance counting pool can be executed in parallel, so as to achieve the dual purposes of increasing the sample measurement speed and reducing the cost and volume of the analyzer.
In the embodiment shown in fig. 2 to 4, in order to further simplify the instrument structure, the fourth control valve (valve V9) for connecting air, which is provided at the upper part of the rear sheath isolation chamber, is also deleted in addition to the existing sheath flow resistance particle analyzer.
Referring to fig. 6, the present invention provides a first liquid level initialization method, which is applied to any one of the sheath flow impedance particle analyzers of fig. 2-4, and the liquid level initialization method includes the following steps:
step S601: the dilution in the sheath fluid bath was emptied.
The positive pressure source 8, the valve V5 and the valve V8 are opened, and the diluent in the sheath fluid reservoir 7 is drained to the waste fluid reservoir 9 through the front reservoir 41 of the sheath flow impedance counter reservoir 4 by the positive pressure, thereby emptying the sheath fluid reservoir 7. Valves V5 and V8 were closed.
Step S602: evacuating liquid from the back sheath isolation tank and the back sheath waste liquid tank;
valve V4 or valve V3 was opened to evacuate the rear sheath isolation tank 6 and rear sheath waste tank 5 using positive pressure.
Since the air control valve is connected to the post-sheath waste liquid tank 5 and the post-sheath isolation tank 6 in the embodiment shown in fig. 2 to 4, the post-sheath isolation tank 6 and the post-sheath waste liquid tank 5 cannot be evacuated by using negative pressure without air.
For this reason, in the present embodiment, it is proposed to evacuate the back sheath separation tank 6 and the back sheath waste liquid tank 5 by using the positive pressure source 8, but the sheath liquid tank 7 has the diluent, and for this purpose, it is necessary to evacuate the diluent of the sheath liquid tank by using the positive pressure source, and then evacuate the back sheath separation tank 6 and the back sheath waste liquid tank 5 by using the positive pressure source.
Step S603: and (4) filling the sheath liquid pool and filling a channel connected with the sheath flow impedance counting pool.
Opening the valve V2 and the valve V8, refilling the sheath fluid reservoir 7 and filling it to recover the sheath fluid emptied in step S601, and filling the sheath fluid channel of the anterior reservoir, the anterior reservoir 41 and the anterior reservoir cleaning channel through the opened valve V2 and the opened valve V8, so as to remove air bubbles during the filling process and avoid the influence of the air bubbles on the sample measurement process.
Step S604: and setting the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool to be preset negative pressure.
In the second embodiment shown in fig. 3, the negative pressure source is set to a predetermined negative pressure, the third control valve V10 between the waste liquid tank 9 and the negative pressure source is opened, and the waste liquid tank 9 is switched to a negative pressure state. In the third embodiment shown in fig. 4, the negative pressure waste liquid pool is set to a preset negative pressure, and the three-way valve is switched to the negative pressure waste liquid pool to be in a negative pressure state.
Then, the valve V7 is opened to set the rear sheath isolation chamber 6, the rear sheath waste liquid pool 5, and the waste liquid pool 9 at a preset negative pressure.
Step S605: and adding a preset amount of diluent into the rear sheath isolation pool through the rear sheath cleaning channel.
Valve V3 is opened and a predetermined amount of diluent is added to the back sheath isolation chamber 6 via the back sheath purge channel. Since the rear sheath isolation chamber 6 is in a negative pressure state, the sheath fluid can be sucked into the rear sheath isolation chamber 6 in a negative pressure state. Since the rear sheath isolation pool 6 is in a negative pressure state, liquid level fluctuation is caused in the liquid adding process.
Step S606: the back sheath channel is set to be in a normal pressure state.
In the second embodiment shown in fig. 3, the third control valve V10 between the waste liquid pool 9 and the negative pressure source is closed, and the waste liquid pool 9 is switched to the normal pressure state. In the third embodiment shown in fig. 4, the three-way valve is switched to the normal pressure waste liquid tank to become the normal pressure state.
Step S607: and after the liquid level in the back sheath isolation pool is stable, finishing the liquid level initialization of the back sheath isolation pool.
And waiting for the liquid level in the back sheath isolation pool 6 to be stable in the normal pressure state, thereby finishing the liquid level initialization of the back sheath isolation pool 6.
In the embodiment shown in fig. 6, the need to drain the sheath fluid reservoir 7 is followed by a return of the fluid level in the sheath fluid reservoir, which consumes a smaller amount of sheath fluid and time to return the sheath fluid reservoir. However, since the liquid level initialization function is not a common function, it is usually used when the instrument is first installed or maintained, and thus, no additional cost is incurred.
On the structure of the sheath flow impedance particle analyzer, a valve V9 on the upper part of the back sheath isolation pool 6 can be eliminated, the cost of the instrument can be obviously saved, the complexity can be reduced, and the device can be used for low-end instruments which are sensitive to the cost of the instrument and have low requirements on the execution time of a function flow.
Referring to fig. 7a-7c, the present invention provides fourth to sixth embodiments of the sheath flow impedance particle analyzer, further comprising, in addition to any one of the embodiments of the sheath flow impedance particle analyzer of fig. 2-4: and a fourth control valve (valve V9) arranged at the upper part of the back sheath isolation pool and communicated with air.
The embodiment of fig. 7a-7c allows the back sheath isolation chamber 6 to be connected to air, and allows the back sheath isolation chamber 6 and the back sheath waste chamber 5 to be more easily emptied during level initialization of the back sheath isolation chamber 6, based on the embodiment of any of the sheath flow impedance particle analyzers of fig. 2-4.
Referring to fig. 8, the present invention provides a second embodiment of a liquid level initialization method, applied to any one of the sheath flow impedance particle analyzers of fig. 7a-7c, the liquid level initialization method comprising the following steps:
step S801: and arranging a rear sheath isolation chamber, a rear sheath waste liquid pool and a waste liquid pool in a negative pressure state, and emptying the liquid in the rear sheath isolation chamber and the rear sheath waste liquid pool.
In the second embodiment shown in fig. 3, the third control valve V10 between the waste liquid pool 9 and the negative pressure source is opened to switch the waste liquid pool 9 to the negative pressure state. In the third embodiment shown in fig. 4, the three-way valve is switched to the negative pressure waste liquid tank to become a negative pressure state.
Then, the valve V7 is opened to place the rear sheath isolation chamber, the rear sheath waste liquid tank and the waste liquid tank in a negative pressure state. And in a negative pressure state, the liquid in the rear sheath isolation chamber 6 and the rear sheath waste liquid pool 5 is emptied by using the negative pressure.
Step S802: setting a rear sheath isolation chamber, a rear sheath waste liquid pool and a waste liquid pool as preset negative pressure;
in the second embodiment shown in fig. 3, the negative pressure source is set to a predetermined negative pressure, the third control valve V10 between the waste liquid tank 9 and the negative pressure source is opened, and the waste liquid tank 9 is switched to a negative pressure state. In the third embodiment shown in fig. 4, the negative pressure waste liquid pool is set to a preset negative pressure, and the three-way valve is switched to the negative pressure waste liquid pool to be in a negative pressure state.
Step S803: adding a preset amount of diluent into the rear sheath isolation pool through a rear sheath cleaning channel;
valve V3 is opened and a predetermined amount of diluent is added to the back sheath isolation chamber 6 via the back sheath purge channel. Since the rear sheath isolation chamber 6 is in a negative pressure state, the sheath fluid can be sucked into the rear sheath isolation chamber 6 in a negative pressure state. Since the rear sheath isolation pool 6 is in a negative pressure state, liquid level fluctuation is caused in the liquid adding process.
Step S804: the rear sheath isolation chamber, the rear sheath waste liquid pool and the waste liquid pool are set to be in a normal pressure state.
In the second embodiment shown in fig. 3, the third control valve V10 between the waste liquid pool 9 and the negative pressure source is closed, and the waste liquid pool 9 is switched to the normal pressure state. In the third embodiment shown in fig. 4, the three-way valve is switched to the normal pressure waste liquid tank to become the normal pressure state.
Step S805: and after the liquid level in the back sheath isolation pool is stable, finishing the liquid level initialization of the back sheath isolation pool.
And waiting for the liquid level in the back sheath isolation pool 6 to be stable in the normal pressure state, thereby finishing the liquid level initialization of the back sheath isolation pool 6.
The first liquid level initialization method embodiment is substantially the same as the second liquid level initialization method embodiment, except that: in the second embodiment, since the third control valve V10 is provided, the rear sheath isolation tank 6 and the rear sheath waste liquid tank 5 can be directly evacuated by using the negative pressure, so that it is not necessary to evacuate the diluent in the sheath liquid tank 7 and evacuate the rear sheath isolation tank 6 and the rear sheath waste liquid tank 5 by using the positive pressure source.
The foregoing is illustrative of embodiments of the present invention, and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the embodiments of the present invention and are intended to be within the scope of the present invention.
Claims (10)
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Cited By (1)
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