CN104914031A - Flow cytometry fluid path system and flow cytometry method - Google Patents
Flow cytometry fluid path system and flow cytometry method Download PDFInfo
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- CN104914031A CN104914031A CN201510324158.9A CN201510324158A CN104914031A CN 104914031 A CN104914031 A CN 104914031A CN 201510324158 A CN201510324158 A CN 201510324158A CN 104914031 A CN104914031 A CN 104914031A
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Abstract
The invention relates to the technical field of human blood detection and discloses a flow cytometry fluid path system. The system consists of a motor drive sample stream injector and a sheath fluid injector, wherein the outlet of the sample stream injector is respectively connected with a diluent bucket and a sample injection port by virtue of a pipeline and a three-way solenoid valve and is also connected with a reaction tank; the outlet of the sheath fluid injector respectively passes through a two-way solenoid valve by virtue of a pipeline and a liquid pressure sensor and is connected with a flow chamber sample injection port, a flow chamber sheath fluid injection port and a fluid suction port of the sample stream injector; a hemolytic agent injector is driven by the motor, and the outlet of the hemolytic agent injector is respectively connected with a hemolytic agent bottle and the reaction tank by virtue of a pipeline and a three-way solenoid valve; the outlet of a sampling injector is connected with a sampling needle by virtue of a pipeline; and a diluent injection port in the sampling injector is connected with the diluent bucket by virtue of a pipeline. The system disclosed by the invention is simple in structure, low in cost and simple and reliable in detection method, the detection data is accurate, and machine miniaturization can be realized.
Description
Technical field
The present invention relates to blood of human body detection technique field, particularly a kind of FCM analysis liquid-way system and FCM analysis method.
Background technology
Sheath Flow Technique is also flow cytometry, is to aim at a kapillary to detect aperture pipe, and cell to be measured or particle form sample stream from kapillary ejection.The sheath fluid (a kind of liquid of clean not disturbed test result) simultaneously allowing kapillary surrounding flow out wraps sample stream and flows fast through detection zone together, and ensures to become single word order between cell or particle are in the detection.FCM analysis liquid-way system is exactly a kind of liquid-way system being applied to medical blood cell analyzer device, is the control carrier realizing FCM analysis.FCM analysis refers to and controls cell by sheath Flow Technique (flow cytometry) or particle becomes single arrangement to complete the fluid control technology of test by detection zone, this technology can avoid cell bonding, the disorder of cell flow trace causes Detection Information to be forbidden through detection zone, the problem of the undetected flase drop of cell.Its sheath stream of existing FCM analysis liquid-way system is formed and mainly contains two kinds of modes: the first is that sheath stream constant voltage source of the gas drives, and sample stream uses syringe to promote.The second is that sheath stream and sample stream use the syringe of separate electrical motor and actuator drives thereof to promote all respectively.The constant voltage source of the gas of the first needs to be equipped with a set of source of the gas and stable-pressure device thereof, system complex, and cost compare is high.Each syringe of the second needs to join separately a set of motor and drive transmission drives, and because the holistic cost of motor and supporting drive transmission is higher, therefore the cost of the program is also higher.
Summary of the invention
The object of this invention is to provide that a kind of structure is simple, cost is low, detection is accurate, can realize the FCM analysis liquid-way system of machine miniaturization and FCM analysis method.
The object of the invention is to be realized by following approach:
FCM analysis liquid-way system, comprises flow chamber 8 and reaction tank 11,
By the sample streamer emitter 2 of motor 1 driving moment balanced arrangement and sheath liquid syringe 3,
Sample streamer emitter 2 exports and is connected dilution bucket 12 and flow chamber sample inlet 7 respectively with three-way solenoid valve V5 by the road, and by the road with pinch valve PV1 coupled reaction pond 11;
Sheath liquid syringe 3 connects sample streamer emitter 2 liquid sucting port after meeting flow chamber sheath fluid inlet 6, connection two-way electromagnetic valve V1 after meeting flow chamber sample inlet 7, connection two-way electromagnetic valve V2 after exporting and connecting two-way electromagnetic valve V3 respectively by the road;
Drive hemolytic agent syringe (13) by motor (14), described hemolytic agent syringe (13) outlet is connected hemolytic agent bottle (15) and reaction tank (11) by the road respectively with three-way solenoid valve V6;
Sampling syringe 18 exports and connects sampling needle 19 by the road, and the dilution inlet 17 on sampling syringe 18 connects dilution bucket 12 by the road.
Further: the sample streamer emitter 2 that described equalising torque is arranged and sheath liquid syringe 3 are that sample streamer emitter 2 and sheath liquid syringe 3 are arranged in motor 1 both sides, equalising torque formula is F1 × S1=F2 × S2, wherein S1 is the distance at the center of sample streamer emitter 2 and motor 1, S2 is the distance at the center of sheath liquid syringe 3 and motor 1, F1 is the promotion resistance of sample streamer emitter 2, and F2 is the promotion resistance of sheath liquid syringe 3.
Further: described motor 14 drives hemolytic agent syringe 13 to be have the mat woven of fine bamboo strips one hemolytic agent syringe 13 and the second hemolytic agent syringe 20, the mat woven of fine bamboo strips one hemolytic agent syringe 13 exports and is connected the mat woven of fine bamboo strips one hemolytic agent bottle 15 and reaction tank 11, second hemolytic agent syringe 20 by the road with three-way solenoid valve V6 respectively and exports and be connected the mat woven of fine bamboo strips two hemolytic agent bottle 21 and reaction tank (11) respectively with three-way solenoid valve V7 by the road.
Further: described sheath liquid syringe (3) outlet serial connection liquid-pressure pick-up LP.
FCM analysis method adopts following steps:
The sampling of the mat woven of fine bamboo strips one step and syringe are for liquid:
Sampling syringe is driven to draw blood sample by sampling needle with motor;
Sample streamer emitter and sheath streamer emitter is driven to draw quantitative dilution in dilution bucket with motor;
The first hemolytic agent syringe and the second hemolytic agent syringe is driven to draw quantitative first hemolytic agent and the second hemolytic agent from the first hemolytic agent bottle and the second hemolytic agent bottle respectively with motor;
The mat woven of fine bamboo strips two step sample incubation reacts:
First blood sample and the first hemolytic agent syringe that sampling needle is drawn being drawn the first quantitative hemolytic agent injects after reaction tank fully reacts, then draws the second quantitative hemolytic agent injection reaction tank with the second hemolytic agent syringe and fully react;
The mat woven of fine bamboo strips three step sample prepares:
By motor driving sheath streamer emitter, sample in reaction tank is drawn into the standby sample line segments of flow chamber sample inlet;
The mat woven of fine bamboo strips four step cell detection:
Sheath liquid syringe and sample streamer emitter is driven with motor, the quantitative dilution that the above-mentioned sheath liquid syringe for liquid is drawn pushes away sheath fluid to flow chamber sheath streamer entrance, the quantitative dilution that sample injections device is drawn pushes away sample stream to flow chamber sample inlet, allow build the pressure in flow chamber stable after open flow chamber outlet again, then sample stream by sheath stream be wrapped in queuing enter flow chamber optical detecting parts carry out analysis detect, with the pressure of liquid-pressure pick-up Real-Time Monitoring sheath streamer emitter, carry out overvoltage protection;
The mat woven of fine bamboo strips five step cleaning systems:
Sheath liquid syringe is driven in dilution bucket, to suck dilution by sample streamer emitter with motor, control the extrapolation of sheath liquid syringe after imbibition in flow chamber, note dilution cleaning, standby sample line segments through flow chamber sample inlet notes dilution cleaning in reaction tank, drives sampling syringe to draw dilution clean sampling needle with motor.
Good effect of the present invention: sample streamer emitter and sheath streamer emitter merge by same motor and actuator drives thereof and adopt equalising torque to arrange by FCM analysis liquid-way system of the present invention and FCM analysis method, not only avoid the sheath dither flow that the difference between different driving mechanism causes, and add integral rigidity and the inertia of system, and then make sheath stream more steady, improve detection accuracy, share motor driver and do not increase other parts in addition and reduce system cost.This invention simplifies the malleation source of the gas of prior art and stable-pressure device thereof and negative pressure source of the gas and stable-pressure device thereof, structure be simple, cost is low, detection method is simple, reliable, detect data accurately, machine miniaturization can be realized.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention.
Fig. 2 is sample streamer emitter of the present invention and sheath liquid syringe equalising torque arrangement schematic diagram.
Fig. 3 is embodiment of the present invention structural representation.
In figure: 1. motor, 2. sample streamer emitter, 3. sheath liquid syringe, 4. three-way connection, 5. three-way connection, 6. flow chamber sheath fluid inlet, 7. flow chamber sample streamer entrance, 8. flow chamber, 9. flow chamber waste liquid outlet, 10. three-way connection, 11. reaction tanks, 12. dilution buckets, 13. first hemolytic agent syringes, 14. motors, 15. first hemolytic agent bottles, 16. motors, 17. dilution inlets, 18. sampling syringe, 19. sampling needles, 20. second hemolytic agent syringes, 21. second hemolytic agent bottles, V1. two three-way electromagnetic valves, V2. two three-way electromagnetic valves, V3. two three-way electromagnetic valves, V4 two three-way electromagnetic valve, V5. three-way solenoid valve, V6 three-way solenoid valve, PV1 pinch valve, LP. liquid-pressure pick-up.
Embodiment
Below in conjunction with Figure of description, the present invention is elaborated:
Embodiment: as Fig. 1, Fig. 2, shown in Fig. 3, FCM analysis liquid-way system of the present invention comprises: sample streamer emitter 2, sheath liquid syringe 3, sampling syringe 18, first hemolytic agent syringe 13, second hemolytic agent syringe 20, sampling needle 19, reaction tank 11, flow chamber 8, dilution bucket 12, first hemolytic agent bottle 15, second hemolytic agent bottle 21, motor 1, motor 16, motor 14, two three-way electromagnetic valve V1, two three-way electromagnetic valve V2, two three-way electromagnetic valve V3, two three-way electromagnetic valve V4, three-way solenoid valve V5, three-way solenoid valve V6, three-way solenoid valve V7, pinch valve PV1, liquid-pressure pick-up LP, three-way connection 4, three-way connection 5, three-way connection 10.Flow chamber 8 is provided with sheath streamer entrance 6, sample streamer entrance 7 and flow chamber waste liquid outlet 9; Flow chamber waste liquid outlet 9 is connected with two three-way electromagnetic valve V4; Sample streamer emitter 2 is exported and is connected with three-way solenoid valve V5 common port by pipeline, and three-way solenoid valve V5 Chang Kaiduan is connected with dilution bucket 12, and three-way solenoid valve V5 normal-closed end is also connected with flow chamber sample inlet 7 with three-way connection 10 by pipeline simultaneously; Sample streamer emitter 2 liquid sucting port to be exported with sheath liquid syringe 3 by pipeline, three-way connection 4 and two three-way electromagnetic valve V1 and is connected, and sheath liquid syringe 3 is also connected with dilution bucket 12 and flow chamber sample inlet 7 by sample streamer emitter 2; Sheath liquid syringe 3 is exported and is also connected with flow chamber sheath fluid inlet 6 with two three-way electromagnetic valve V2 by pipeline, three-way connection 4, three-way connection 5; Sheath liquid syringe 3 is exported and is connected with flow chamber sample streamer entrance 7 by pipeline, liquid-pressure pick-up LP, three-way connection 4, three-way connection 5 and two three-way electromagnetic valve V3, and flow chamber sample streamer entrance 7 is connected with reaction tank 11 with pinch valve PV1 by pipeline, three-way connection 10 simultaneously; Sampling syringe 18 is connected with sampling needle 19 by pipeline, and arranges dilution inlet 17 on sampling syringe 18; First hemolytic agent syringe 13 is connected with three-way solenoid valve V6 common port by pipeline, and three-way solenoid valve V6 normal-closed end is connected with reaction tank 11, and three-way solenoid valve V6 Chang Kaiduan is connected with the first hemolytic agent bottle 15.Second hemolytic agent syringe 20 is connected with three-way solenoid valve V7 common port by pipeline, and three-way solenoid valve V7 normal-closed end is connected with reaction tank 11, and three-way solenoid valve V7 Chang Kaiduan is connected with the second hemolytic agent bottle 21.Sample streamer emitter 2 and sheath liquid syringe 3 common motor 1 drive; Sampling syringe 18 is driven by motor 16; First hemolytic agent syringe 13 and the second hemolytic agent syringe 20 merge and are driven by motor 14.In FCM analysis liquid-way system, sample streamer is penetrated 2 and is adopted equalising torque to arrange with sheath liquid syringe 3, as shown in Figure 2, equalising torque formula F 1 × S1=F2 × S2, wherein S1 is the distance at the center of sample streamer emitter 2 and motor 1, S2 is the distance at the center of sheath liquid syringe 3 and motor 1, and F1 is the promotion resistance of sample streamer emitter 2, and F2 is the promotion resistance of sheath liquid syringe 3.Sheath liquid syringe (3) outlet serial connection liquid-pressure pick-up LP, Monitoring and prevention overpressure is too high.
FCM analysis method of the present invention adopts following steps:
The sampling of the mat woven of fine bamboo strips one step and syringe are for liquid: motor 16 drives sampling syringe 18 to draw blood sample by sampling needle 19.While suction sample, motor 1 drives sample streamer emitter 2 and sheath streamer emitter 3, open two three-way electromagnetic valve V1, make two syringes all in dilution bucket 12, draw quantitative dilution, motor 14 drives the first hemolytic agent syringe 13 to draw quantitatively the first hemolytic agent from the first hemolytic agent bottle 15.
Second step sample incubation reacts: three-way solenoid valve V6 powers on, and motor 14 drives the first hemolytic agent syringe 13 that quantitative first hemolytic agent is joined reaction tank 11.Simultaneously sampling needle 19 moves to reaction tank 11 and distributes quantitative blood sample and react to reaction tank 11 and the first hemolytic agent, after sample and the first hemolytic agent react setting-up time, three-way solenoid valve V7 powers on, and motor 14 drives the second hemolytic agent syringe 20 quantitative second hemolytic agent to be joined reaction tank 11 and sample continues to react.
3rd step sample prepares: after sample and hemolytic agent are hatched, sheath streamer emitter 13 is driven sample in reaction tank 11 to be drawn into the standby sample line segments of flow chamber sample inlet 7 with motor 1, two three-way electromagnetic valve V5 power on, motor 1 drives sheath streamer emitter to inhale for 3 times, sample is sucked from reaction tank 11 pinch valve PV1 in the standby sample pipeline of flow chamber sample inlet 7, namely exceed inlet 7 certain distance and make standby sample.
4th step cell detection: after standby sample is complete, two three-way electromagnetic valve V2 and three-way solenoid valve V5 power on, motor 1 drives in sheath liquid syringe 3 and sample streamer emitter 2 and pushes away, sheath streamer emitter 3 pushes away sheath fluid to flow chamber sheath streamer entrance 6, sample injections device 2 pushes away sample stream to flow chamber sample inlet 7, two syringes push away simultaneously certain hour by build the pressure in flow chamber 8 stable after open flow chamber again and export two three-way electromagnetic valve V4, then sample stream will flow out by sheath stream be wrapped in make cell queue up enter flow chamber 8 optical detecting parts carry out analysiss detection, with the pressure of liquid-pressure pick-up LP Real-Time Monitoring sheath streamer emitter in whole process, carry out overvoltage protection.
5th step cleaning systems: after cell detection terminates, two three-way electromagnetic valve V1 power on, motor 1 drives sheath liquid syringe 3 in dilution bucket 12, to suck dilution by sample streamer emitter 2, after imbibition, sheath liquid syringe 3 is extrapolated, open two three-way electromagnetic valve V2 and can inject dilution in flow chamber 8, cleaning flow chamber 8, open two three-way electromagnetic valve V3 and can clean the standby sample pipeline that the specimen needle in flow chamber 8 is connected with flow chamber sample inlet 7, dilution flows into reaction tank 11 by pinch valve PV1, can rinse again to reaction tank 11.Motor 16 drives sampling syringe 18 to draw dilution and can clean sampling needle 19.After cleaning, all parts return to measurement standby condition.
The present invention by sample streamer emitter and sheath streamer emitter by same motor and actuator drives thereof, and adopt equalising torque to arrange, not only avoid the sheath dither flow that the difference between different driving mechanism causes, and add integral rigidity and the inertia of system, and then make sheath stream more steady, improve detection accuracy, share motor driver and do not increase other parts in addition, reduce system cost, sampling needle does not connect valve in the middle of being directly connected with sampling syringe simultaneously, ensure that the accuracy of blood sampling point blood, so be a kind of more accurately cheaper FCM analysis liquid-way system.Sample streamer emitter and sheath liquid syringe is driven with same motor, because the action of sample streamer emitter is little, and when forming sheath stream, sample streamer emitter and sheath streamer emitter need to move simultaneously, need the task of sample streamer emitter single movement also rationally can have been arranged by sheath liquid syringe in analytic process, the present invention allows under sample streamer emitter default conditions directly leads to dilution bucket, so sheath liquid syringe is executed the task separately, the sample streamer emitter interlock produced there is no impact to system works, and therefore test the speed also can not be influenced for complete machine.
Claims (5)
1. a FCM analysis liquid-way system, comprises flow chamber (8) and reaction tank (11), it is characterized in that
By sample streamer emitter (2) and the sheath liquid syringe (3) of motor (1) driving moment balanced arrangement,
The outlet of described sample streamer emitter (2) is connected dilution bucket (12) and flow chamber sample inlet (7) by the road respectively with three-way solenoid valve V5, and by the road with pinch valve PV1 coupled reaction pond (11);
Sample streamer emitter (2) liquid sucting port is connect after meeting flow chamber sheath fluid inlet (6), connection two-way electromagnetic valve V1 after meeting flow chamber sample inlet (7), connection two-way electromagnetic valve V2 after described sheath liquid syringe (3) outlet connects two-way electromagnetic valve V3 by the road respectively;
Drive hemolytic agent syringe (13) by motor (14), described hemolytic agent syringe (13) outlet is connected hemolytic agent bottle (15) and reaction tank (11) by the road respectively with three-way solenoid valve V6;
Sampling syringe (18) outlet connects sampling needle (19) by the road, and the dilution inlet (17) on sampling syringe (18) connects dilution bucket (12) by the road.
2. FCM analysis liquid-way system according to claim 1, it is characterized in that the sample streamer emitter (2) that described equalising torque is arranged and sheath liquid syringe (3) are that sample streamer emitter (2) and sheath liquid syringe (3) are arranged in motor (1) both sides, equalising torque formula is F1 × S1=F2 × S2, wherein S1 is the distance at the center of sample streamer emitter (2) and motor (1), S2 is the distance at the center of sheath liquid syringe (3) and motor (1), F1 is the promotion resistance of sample streamer emitter (2), F2 is the promotion resistance of sheath liquid syringe (3).
3. FCM analysis liquid-way system according to claim 1, it is characterized in that described motor (14) drives hemolytic agent syringe (13) to be have the mat woven of fine bamboo strips one hemolytic agent syringe (13) and the second hemolytic agent syringe (20), the mat woven of fine bamboo strips one hemolytic agent syringe (13) outlet is connected the mat woven of fine bamboo strips one hemolytic agent bottle (15) and reaction tank (11) by the road respectively with three-way solenoid valve V6, the second hemolytic agent syringe (20) outlet is connected the mat woven of fine bamboo strips two hemolytic agent bottle (21) and reaction tank (11) by the road respectively with three-way solenoid valve V7.
4. FCM analysis liquid-way system according to claim 1, is characterized in that described sheath liquid syringe (3) outlet serial connection liquid-pressure pick-up LP.
5. a FCM analysis method, is characterized in that adopting following steps:
The sampling of the mat woven of fine bamboo strips one step and syringe are for liquid:
Sampling syringe is driven to draw blood sample by sampling needle with motor;
Sample streamer emitter and sheath streamer emitter is driven to draw quantitative dilution in dilution bucket with motor;
The first hemolytic agent syringe and the second hemolytic agent syringe is driven to draw quantitative first hemolytic agent and the second hemolytic agent from the first hemolytic agent bottle and the second hemolytic agent bottle respectively with motor;
The mat woven of fine bamboo strips two step sample incubation reacts:
First blood sample and the first hemolytic agent syringe that sampling needle is drawn being drawn the first quantitative hemolytic agent injects after reaction tank fully reacts, then draws the second quantitative hemolytic agent injection reaction tank with the second hemolytic agent syringe and fully react;
The mat woven of fine bamboo strips three step sample prepares:
By motor driving sheath streamer emitter, sample in reaction tank is drawn into the standby sample line segments of flow chamber sample inlet;
The mat woven of fine bamboo strips four step cell detection:
Sheath liquid syringe and sample streamer emitter is driven with motor, the quantitative dilution that the above-mentioned sheath liquid syringe for liquid is drawn pushes away sheath fluid to flow chamber sheath streamer entrance, the quantitative dilution that sample injections device is drawn pushes away sample stream to flow chamber sample inlet, allow build the pressure in flow chamber stable after open flow chamber outlet again, then sample stream by sheath stream be wrapped in queuing enter flow chamber optical detecting parts carry out analysis detect, with the pressure of liquid-pressure pick-up Real-Time Monitoring sheath streamer emitter, carry out overvoltage protection;
The mat woven of fine bamboo strips five step cleaning systems:
Sheath liquid syringe is driven in dilution bucket, to suck dilution by sample streamer emitter with motor, control the extrapolation of sheath liquid syringe after imbibition in flow chamber, note dilution cleaning, standby sample line segments through flow chamber sample inlet notes dilution cleaning in reaction tank, drives sampling syringe to draw dilution clean sampling needle with motor.
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