CN113654972B - Flow cell detection liquid path system and flow cell detection method - Google Patents
Flow cell detection liquid path system and flow cell detection method Download PDFInfo
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- CN113654972B CN113654972B CN202111007429.XA CN202111007429A CN113654972B CN 113654972 B CN113654972 B CN 113654972B CN 202111007429 A CN202111007429 A CN 202111007429A CN 113654972 B CN113654972 B CN 113654972B
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- 239000007788 liquid Substances 0.000 title claims abstract description 134
- 238000001514 detection method Methods 0.000 title claims abstract description 81
- 239000006285 cell suspension Substances 0.000 claims abstract description 52
- 230000000694 effects Effects 0.000 claims abstract description 10
- 238000000684 flow cytometry Methods 0.000 claims abstract description 10
- 238000002835 absorbance Methods 0.000 claims description 11
- 230000003749 cleanliness Effects 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 11
- 230000003204 osmotic effect Effects 0.000 claims description 11
- 239000012530 fluid Substances 0.000 claims description 9
- 239000002245 particle Substances 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 238000004458 analytical method Methods 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 230000006835 compression Effects 0.000 claims description 3
- 238000007906 compression Methods 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 230000037361 pathway Effects 0.000 claims 5
- 238000007822 cytometric assay Methods 0.000 claims 2
- 239000006194 liquid suspension Substances 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 40
- 238000009434 installation Methods 0.000 description 7
- 238000012423 maintenance Methods 0.000 description 7
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- 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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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
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- 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
- G01N15/1434—Optical arrangements
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- 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
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
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- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
- G01N2015/1472—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle with colour
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Abstract
The invention discloses a flow type cell detection liquid path system, and relates to the field of cell detection. The flow cell detection liquid path system comprises a flow chamber outer shell, a cell suspension conveying pipe and a sheath liquid conveying pipe. This flow cytometry detection liquid way system is equipped with sheath liquid quality detection box body in the flow cytometry detection liquid way system that sets up when using, sheath liquid quality detection box body can detect the inside sheath liquid of sheath liquid conveyer pipe, reduce the inaccuracy of detection data, this liquid way system overall structure is comparatively ripe, the mixing effect of sheath liquid and cell suspension can be guaranteed to the hybrid tube that sets up, after sheath liquid receives pressure to get into the hybrid tube, sheath liquid conveyer pipe and hybrid tube are inclination, therefore sheath liquid produces the vortex, and finally the downward movement, form the sheath flow with the cell suspension mixture that cell suspension conveyer pipe bottom was carried, cell suspension is located central point, make the sheath liquid wrap up around the cell suspension.
Description
Technical Field
The invention relates to the technical field of cell detection, in particular to a flow cell detection liquid path system and a flow cell detection method.
Background
The sheath flow technique is also called flow cytometry, which is to use a capillary tube to align with a detection small hole tube, the cells or particles to be detected are ejected from the capillary tube to form a sample flow, and sheath liquid (a liquid which is clean and does not interfere with the test result) flowing out of the periphery of the capillary tube wraps the sample flow to flow through a detection area together and ensures that the cells or particles are arranged in a single line in the middle of the detection area, and the flow type cell detection liquid path system is a liquid path system applied to a medical blood cell analysis instrument and is a control carrier for realizing flow type cell detection.
In the prior art, in a liquid path system for detecting flow-type cells, cleanliness, pH value, absorbance and osmotic pressure of sheath liquid cannot be detected, so that in the prior art, the phenomenon that detection data are inaccurate due to the fact that one or more indexes of the sheath liquid are not up to standard often occurs in the using process of the sheath liquid, the data have great influence on detection results, the existing detection method is imperfect, in the whole cell detection process, the problem that detection information is inaccurate due to the fact that cell flow track is disordered when the traditional sheath liquid and cell suspension are mixed finally enter a laser detection pipeline is solved, through long-term experiments, most of ultrasonic vibrators arranged at a nozzle at the bottom of a flow chamber shell are damaged, in the case that the ultrasonic vibrators are damaged, parameter changes easily due to long-term high-frequency vibration in the working process of the ultrasonic vibrators, and therefore periodic maintenance and replacement are needed, in the prior art, the problem that later maintenance is inconvenient is caused by the ultrasonic vibrators arranged in the prior art is solved, and the method for detecting flow-type cell detection is disclosed in the prior art.
Disclosure of Invention
(One) solving the technical problems
Aiming at the defects of the prior art, the invention discloses a flow cell detection liquid path system and a flow cell detection method, which are used for solving the problems in the background art.
(II) technical scheme
In order to achieve the above purpose, the invention is realized by the following technical scheme: a flow cytometry detection fluid path system comprising:
The device comprises a flow outdoor shell, wherein a reinforced fixed nozzle pipe is arranged at the bottom of the flow outdoor shell, a vibrating pipe is connected inside the reinforced fixed nozzle pipe in a sliding manner, an ultrasonic vibrator is arranged at the bottom of the flow outdoor shell, a vibrating head is arranged at the bottom of the ultrasonic vibrator, and the vibrating head is fixedly connected with the vibrating pipe;
the cell suspension conveying pipe is fixedly connected to the top of the outdoor shell of the flow chamber, the outer wall of the cell suspension conveying pipe is fixedly connected with the mixing pipe, and the mixing pipe is arranged in the inner cavity of the outdoor shell of the flow chamber;
Sheath liquid conveyer pipe, sheath liquid conveyer pipe fixed connection is in the side of the outdoor casing of flow, sheath liquid conveyer pipe passes the outdoor casing of flow and fixedly links to each other with the outer wall of hybrid tube, fixedly connected with sheath liquid quality detection box body on the sheath liquid conveyer pipe, be equipped with cleanliness factor sensor, PH value sensor, absorbance sensor and osmotic pressure sensor on the sheath liquid quality detection box body.
Preferably, the upper end of the mixing tube is fixedly connected with a plurality of groups of array distributed fixing strips, and the array distributed fixing strips are fixedly connected to the outer wall of the cell suspension conveying tube.
Preferably, the bottom fixedly connected with fixed base of outdoor casing flows, ultrasonic vibrator fixed connection is in fixed base's below, fixed base's bottom fixedly connected with spacing post, ultrasonic vibrator's both sides fixedly connected with limit sleeve, limit sleeve activity cup joints on spacing post.
Preferably, the clamping grooves are formed in two sides of the ultrasonic vibrator, the vertical plate is fixedly connected to the bottom of the fixing base, the hollow square column is fixedly connected to the side wall of the vertical plate, the square positioning sliding block is slidingly connected to the inside of the hollow square column, the first pressure spring is fixedly connected to one side of the square positioning sliding block, the wedge-shaped clamping fixing block is fixedly connected to the other side of the square positioning sliding block, and the wedge-shaped clamping fixing block penetrates through the hollow square column and the vertical plate and is movably clamped in the clamping grooves.
Preferably, the bottom of square location slider fixedly connected with handle, the fluting that corresponds with square location slider handle is seted up to the bottom of cavity square column.
Preferably, the lateral wall fixedly connected with second connecting block of vibrations head, the lateral wall fixedly connected with chucking piece of counterpointing of second connecting block, the lateral wall fixedly connected with first connecting block of vibration pipe, the lateral wall fixedly connected with U-shaped cassette of first connecting block, U-shaped cassette and chucking piece joint of counterpointing, the spout that corresponds with first connecting block has been seted up to the lateral wall of the fixed nozzle pipe of strenghthened type.
Preferably, the side walls of the U-shaped clamping seat and the alignment clamping block are movably inserted with a U-shaped clamping piece, and two groups of round holes corresponding to the U-shaped clamping piece are formed in the side walls of the U-shaped clamping seat and the alignment clamping block.
Preferably, the inside of counterpoint chucking piece is equipped with the cylinder groove, sliding connection has the slip plectane in the cylinder groove of counterpoint chucking piece, the bottom fixedly connected with locking lever of slip plectane, the locking lever passes counterpoint chucking piece and activity grafting in U-shaped chucking spare, the round hole that corresponds with the locking lever has been seted up on the U-shaped chucking spare, the top fixedly connected with second compression spring and the pull rod of slip plectane, the top of pull rod passes counterpoint chucking piece and upwards extends.
Preferably, the detection method comprises the following steps:
step S1: preparing cells or particles to be tested into a cell suspension, and then pressing a sample to be tested into a cell suspension conveying pipe under a certain pressure;
step S2: conveying the sheath liquid by a cell suspension conveying pipe, detecting the sheath liquid when the sheath liquid passes through a sheath liquid quality detection box body, and directly pressurizing and conveying the sheath liquid into the mixing pipe when the cleanliness, the PH value, the absorbance and the osmotic pressure of the sheath liquid are normal;
Step S3: when the cell suspension moves downwards, the cell suspension is finally mixed with the sheath liquid to form mixed liquid, the sheath liquid is wrapped outside the cell suspension for protection under the action of pressure, and the mixed liquid moves downwards and passes through the inner cavity of the outer shell of the flow chamber and is sprayed out by the reinforced fixed nozzle pipe;
step S4: when passing through the reinforced fixed nozzle pipe, the vibrating pipe is continuously vibrated by the ultrasonic vibrator to move up and down, the mixed liquid is broken into uniform liquid drops, cells to be detected are dispersed in the liquid drops, and finally the liquid drops are injected into the laser detection pipeline;
Step S5: when the fluorescent marker passes through the laser irradiation area, fluorescent signals representing different substances and different wavelengths inside the cell are generated, and the signals are emitted to a spatial omnibearing solid angle by taking the cell as the center to generate scattered light and fluorescent signals;
Step S6: the fluorescence generated by the fluorescence-dyed cells after being excited by the appropriate light waves is converted into an electric signal through a photoelectric converter, and the output electric signal is amplified and then transmitted to a computer, and finally data processing and analysis are carried out, so that the detection of the cells is formed.
The invention discloses a flow cell detection liquid path system, which has the following beneficial effects:
1. The flow type cell detection liquid path system is characterized in that a sheath liquid quality detection box body is arranged in the flow type cell detection liquid path system when the flow type cell detection liquid path system is used, the sheath liquid quality detection box body can detect sheath liquid in the sheath liquid conveying pipe, the sheath liquid can be monitored by a cleanliness sensor, a PH value sensor, an absorbance sensor and an osmotic pressure sensor, when certain data are abnormal, the detection process can be alarmed and prevented by a computer, the inaccuracy of the detection data is reduced, the whole structure of the flow type cell detection liquid path system is mature, the mixing pipe can ensure that the mixing effect of the sheath liquid and the cell suspension is good when the flow type cell detection liquid path system is used, after the sheath liquid is subjected to pressure, the sheath liquid conveying pipe and the mixing pipe are inclined, so that vortex is generated by the sheath liquid, and finally moves downwards, the sheath liquid is mixed with the cell suspension conveyed by the bottom of the cell suspension conveying pipe to form sheath liquid, and the cell suspension is located at the center position, the periphery of the cell suspension is wrapped, the better protection is formed, and the use process of the flow path system is ensured to be more perfect.
2. This flow cytometry liquid way system, when using, because ultrasonic vibrator's life is shorter relatively, belong to the wearing parts, consequently in the later maintenance, structural design in the device is utilized for ultrasonic vibrator conveniently fixes in the below of the outdoor casing of flow, and be convenient for fix with the vibrating tube, ultrasonic vibrator conveniently carries out the dismouting in the later stage, convenient maintenance, guarantee the normal work of vibrating tube, consequently, make the fixed nozzle pipe of strenghthened type carry out the blowout of mixed liquid, can be continuous with mixed liquid fracture for even liquid drop, guarantee to treat the processing of survey cell good, guarantee the stability of device, guarantee the detection effect.
3. This flow cell detection liquid way system, the cooperation of spacing post and the stop collar that set up has been realized carrying out ultrasonic vibrator's installation fixedly on the one hand, realize the installation location to ultrasonic vibrator, on the other hand realized that ultrasonic vibrator installs fixedly the back, ultrasonic vibrator receives the side direction to support, guarantee stably, be equipped with the inclined plane on the wedge chucking fixed block that sets up, therefore when installing ultrasonic vibrator, the operation is simpler, direct with spacing post and stop collar alignment cup joint when ultrasonic vibrator installs, and promote ultrasonic vibrator chucking can, when ultrasonic vibrator offsets with the inclined plane of wedge chucking fixed block, can promote the automatic shrinkage of wedge chucking fixed block in the inner chamber of cavity square post, and ultrasonic vibrator is when promoting fixed position department, wedge chucking fixed block receives the outside spring again, and insert to the chucking inslot and form the fixing to ultrasonic vibrator, the installation step has been simplified.
Drawings
FIG. 1 is a perspective view of a first perspective view of a three-dimensional structure of the present invention;
FIG. 2 is a perspective view of a second perspective view of the stereographic structure of the present invention;
FIG. 3 is a cross-sectional view of the present invention at a first view angle;
FIG. 4 is a cross-sectional view of the present invention at a second perspective;
FIG. 5 is a bottom view of the flow outdoor housing of the present invention;
FIG. 6 is an internal structural view of the reinforced fixed nozzle pipe of the present invention;
FIG. 7 is a block diagram of an alignment clamp block of the present invention;
FIG. 8 is a cross-sectional view of the alignment clamping block of the present invention;
FIG. 9 is a block diagram of a stationary base of the present invention;
FIG. 10 is an exploded view of the shock tube and reinforced fixed nozzle tube of the present invention;
FIG. 11 is an exploded view of the hollow square column and square positioning slide of the present invention.
In the figure: 1. a flow outdoor housing; 2. a cell suspension delivery tube; 3. a sheath fluid delivery tube; 4. a mixing tube; 401. an array distributed fixing strip; 5. sheath fluid quality detection box body; 6. a cleanliness sensor; 7. a pH sensor; 8. an absorbance sensor; 9. an osmotic pressure sensor; 10. a reinforced fixed nozzle tube; 11. a laser detection pipeline; 12. a vibrating tube; 13. an ultrasonic vibrator; 1301. a limit sleeve; 1302. a clamping groove; 14. a vibrating head; 15. a fixed base; 16. a limit column; 17. a vertical plate; 18. a hollow square column; 19. square positioning slide block; 20. a first pressure spring; 21. wedge-shaped clamping fixing blocks; 22. a first connection block; 2201. a chute; 23. a U-shaped clamping seat; 24. a second connection block; 25. aligning and clamping blocks; 26. a U-shaped clamping piece; 27. sliding the circular plate; 28. a locking lever; 29. a second pressure spring; 30. and (5) a pull rod.
Detailed Description
Example 1:
The embodiment of the invention discloses a flow cell detection liquid path system, as shown in figures 1-11, comprising:
The flow chamber comprises a flow chamber housing 1, wherein a reinforced fixed nozzle pipe 10 is arranged at the bottom of the flow chamber housing 1, a vibrating pipe 12 is connected in a sliding manner in the reinforced fixed nozzle pipe 10, an ultrasonic vibrator 13 is arranged at the bottom of the flow chamber housing 1, a vibrating head 14 is arranged at the bottom of the ultrasonic vibrator 13, and the vibrating head 14 is fixedly connected with the vibrating pipe 12;
The cell suspension conveying pipe 2 is fixedly connected to the top of the flow outdoor shell 1, the outer wall of the cell suspension conveying pipe 2 is fixedly connected with the mixing pipe 4, and the mixing pipe 4 is arranged in the inner cavity of the flow outdoor shell 1;
sheath liquid conveyer pipe 3, sheath liquid conveyer pipe 3 fixed connection is in the side of the outdoor casing 1 of flow, sheath liquid conveyer pipe 3 pass the outdoor casing 1 of flow and fixedly link to each other with the outer wall of hybrid tube 4, fixedly connected with sheath liquid quality detection box body 5 on the sheath liquid conveyer pipe 3, be equipped with cleanliness factor sensor 6, PH value sensor 7, absorbance sensor 8 and osmotic pressure sensor 9 on the sheath liquid quality detection box body 5.
According to fig. 4, the upper end fixedly connected with multiunit array distributing type fixed strip 401 of hybrid tube 4, array distributing type fixed strip 401 fixed connection is on the outer wall of cell suspension conveyer pipe 2, and multiunit array distributing type fixed strip 401 that sets up has realized the fixed to hybrid tube 4, and hybrid tube 4 is fixed on the outer wall of cell suspension conveyer pipe 2 and with cell suspension conveyer pipe 2 contactless, cell suspension conveyer pipe 2 and the axis coincidence of hybrid tube 4, and the hybrid tube 4 of setting can guarantee that cell suspension and sheath liquid can better mix and form the sheath flow.
The detection method comprises the following steps:
step S1: preparing cells or particles to be tested into a cell suspension, and then pressing a sample to be tested into the cell suspension conveying pipe 2 under a certain pressure;
step S2: the sheath liquid is conveyed by a cell suspension conveying pipe 2, the sheath liquid is detected when passing through a sheath liquid quality detection box body 5, and the sheath liquid is directly pressurized and conveyed into the mixing pipe 4 when the cleanliness, the PH value, the absorbance and the osmotic pressure of the sheath liquid are normal;
Step S3: when the cell suspension moves downwards, the cell suspension is finally mixed with the sheath liquid to form a mixed liquid, the sheath liquid is wrapped outside the cell suspension for protection under the action of pressure, and the mixed liquid moves downwards and passes through the inner cavity of the flow chamber outer shell 1 and is sprayed out by the reinforced fixed nozzle pipe 10;
step S4: when passing through the reinforced fixed nozzle pipe 10, the vibrating pipe 12 is continuously vibrated by the ultrasonic vibrator 13 to move up and down, the mixed liquid is broken into uniform liquid drops, cells to be detected are dispersed in the liquid drops, and finally the liquid drops are injected into the laser detection pipeline 11;
Step S5: when passing through the laser detection pipeline 11, the side surface of the laser detection pipeline 11 is provided with a laser source, the focused light beam vertically irradiates on the mixed liquid, when cells carry fluorescein markers to pass through a laser irradiation area, fluorescent signals representing different substances and different wavelengths in the cells are generated, and the signals are emitted to a spatial omnibearing solid angle by taking the cells as the center to generate scattered light and fluorescent signals;
Step S6: the fluorescence generated by the fluorescence-dyed cells after being excited by the appropriate light waves is converted into an electric signal through a photoelectric converter, and the output electric signal is amplified and then transmitted to a computer, and finally data processing and analysis are carried out, so that the detection of the cells is formed.
When the device is used, the sheath liquid quality detection box body 5 is arranged in the flow type cell detection liquid path system, the sheath liquid quality detection box body 5 can detect sheath liquid in the sheath liquid conveying pipe 3, the sheath liquid is guaranteed to be monitored by the cleanliness sensor 6, the PH value sensor 7, the absorbance sensor 8 and the osmotic pressure sensor 9, when one item of data is abnormal, the detection process can be warned and prevented by a computer, the inaccuracy of the detection data is reduced, the whole structure of the liquid path system is mature, the mixing pipe 4 can ensure that the mixing effect of the sheath liquid and the cell suspension is good when the liquid path system is used, after the sheath liquid is pressurized, the sheath liquid conveying pipe 3 and the mixing pipe 4 are inclined, so that the sheath liquid generates vortex and finally moves downwards, the sheath liquid is mixed with the cell suspension conveyed by the bottom of the cell suspension conveying pipe 2 to form sheath flow, and the cell suspension is located at the center position, the periphery of the cell suspension is wrapped, and the use process of the liquid path system is guaranteed to be more perfect.
Example 2:
The embodiment of the invention discloses a flow cell detection liquid path system, as shown in fig. 1-11, comprising:
The flow chamber comprises a flow chamber housing 1, wherein a reinforced fixed nozzle pipe 10 is arranged at the bottom of the flow chamber housing 1, a vibrating pipe 12 is connected in a sliding manner in the reinforced fixed nozzle pipe 10, an ultrasonic vibrator 13 is arranged at the bottom of the flow chamber housing 1, a vibrating head 14 is arranged at the bottom of the ultrasonic vibrator 13, and the vibrating head 14 is fixedly connected with the vibrating pipe 12;
The cell suspension conveying pipe 2 is fixedly connected to the top of the flow outdoor shell 1, the outer wall of the cell suspension conveying pipe 2 is fixedly connected with the mixing pipe 4, and the mixing pipe 4 is arranged in the inner cavity of the flow outdoor shell 1;
sheath liquid conveyer pipe 3, sheath liquid conveyer pipe 3 fixed connection is in the side of the outdoor casing 1 of flow, sheath liquid conveyer pipe 3 pass the outdoor casing 1 of flow and fixedly link to each other with the outer wall of hybrid tube 4, fixedly connected with sheath liquid quality detection box body 5 on the sheath liquid conveyer pipe 3, be equipped with cleanliness factor sensor 6, PH value sensor 7, absorbance sensor 8 and osmotic pressure sensor 9 on the sheath liquid quality detection box body 5.
According to fig. 5, 7 and 10, the bottom fixedly connected with fixed base 15 of outdoor casing 1 flows, ultrasonic vibrator 13 fixedly connected with is in the below of fixed base 15, the bottom fixedly connected with spacing post 16 of fixed base 15, the both sides fixedly connected with spacing sleeve 1301 of ultrasonic vibrator 13, spacing sleeve 1301 activity cup joints on spacing post 16, and the cooperation of spacing post 16 and spacing sleeve 1301 that sets up has realized when carrying out the installation fixation of ultrasonic vibrator 13 on the one hand, realizes the installation location to ultrasonic vibrator 13 on the other hand has realized that after the installation is fixed to ultrasonic vibrator 13, ultrasonic vibrator 13 receives the side direction support, guarantees stably.
According to fig. 5, 7, 10 and 11, the clamping groove 1302 is formed on two sides of the ultrasonic vibrator 13, the vertical plate 17 is fixedly connected to the bottom of the fixing base 15, the hollow square column 18 is fixedly connected to the side wall of the vertical plate 17, the square positioning slide block 19 is slidably connected to the inside of the hollow square column 18, the first pressure spring 20 is fixedly connected to one side of the square positioning slide block 19, the wedge clamping fixing block 21 is fixedly connected to the other side of the square positioning slide block 19, the wedge clamping fixing block 21 passes through the hollow square column 18 and the vertical plate 17 and is movably clamped in the clamping groove 1302, and an inclined plane is formed on the wedge clamping fixing block 21, so that when the ultrasonic vibrator 13 is installed, the operation is simpler, the limit column 16 and the limit sleeve 1301 are directly aligned and sleeved, and the ultrasonic vibrator 13 is pushed to clamp, when the ultrasonic vibrator 13 and the inclined plane of the wedge clamping fixing block 21 are pushed, the wedge fixing block 21 is automatically contracted into the inner cavity of the hollow square column 18, and when the ultrasonic vibrator 13 is pushed to the fixing block 21 is pushed to the fixing position, the wedge clamping spring 21 is pushed out, and the first pressure spring 20 is pushed to the outside the wedge clamping fixing block is ejected to the clamping groove 1302, and the step of the ultrasonic vibrator is formed, and the ultrasonic vibrator is mounted to be clamped and the ultrasonic vibrator is ejected.
According to fig. 10 and 11, the bottom of the square positioning slide block 19 is fixedly connected with a handle, a slot corresponding to the handle of the square positioning slide block 19 is formed in the bottom of the hollow square column 18, and the set handle facilitates the user to slide the square positioning slide block 19, so that the wedge-shaped clamping fixed block 21 is conveniently controlled to move.
According to fig. 6, 7, 9 and 10, the lateral wall fixedly connected with second connecting block 24 of vibrations head 14, the lateral wall fixedly connected with chucking piece 25 of counterpoint of second connecting block 24, the lateral wall fixedly connected with first connecting block 22 of vibration pipe 12, the lateral wall fixedly connected with U-shaped cassette 23 of first connecting block 22, U-shaped cassette 23 and counterpoint chucking piece 25 joint, spout 2201 that corresponds with first connecting block 22 has been seted up to the lateral wall of the fixed nozzle pipe 10 of strenghthened type, counterpoint chucking piece 25 and the "U" shape bayonet socket lock of U-shaped cassette 23 of setting.
According to fig. 6, 7, 9 and 10, the side walls of the U-shaped clamping seat 23 and the alignment clamping block 25 are movably inserted with a U-shaped clamping piece 26, two groups of round holes corresponding to the U-shaped clamping piece 26 are formed in the side walls of the U-shaped clamping seat 23 and the alignment clamping block 25, the set U-shaped clamping seat 23 and the alignment clamping block 25 are limited mainly through the U-shaped clamping piece 26, and two groups of inserting columns are arranged on the U-shaped clamping piece 26 and inserted into the round holes formed in the side walls of the U-shaped clamping seat 23 and the alignment clamping block 25.
According to fig. 5, 7 and 8, the inside of counterpoint chucking piece 25 is equipped with the cylinder groove, sliding connection has a slip plectane 27 in the cylinder inslot of counterpoint chucking piece 25, the bottom fixedly connected with locking lever 28 of slip plectane 27, locking lever 28 passes counterpoint chucking piece 25 and activity grafting in U-shaped chucking piece 26, the round hole that corresponds with locking lever 28 has been seted up on the U-shaped chucking piece 26, the top fixedly connected with second compression spring 29 and pull rod 30 of slip plectane 27, the top of pull rod 30 passes counterpoint chucking piece 25 and upwards extends, and the pull rod 30 that sets up makes things convenient for the user to upwards pull slip plectane 27, controls locking lever 28 motion promptly, consequently conveniently utilizes locking lever 28 to fix and pull down U-shaped chucking piece 26, and U-shaped chucking piece 26 forms the fixed to U-shaped cassette 23 and counterpoint chucking piece 25.
The ultrasonic vibrator 13 that sets up is convenient to be dismantled and assembled, therefore the later maintenance is convenient, and convenient quick changes the operation to ultrasonic vibrator 13 when ultrasonic vibrator 13 damages, guarantees the normal use of device.
When the ultrasonic vibrator 13 is installed and fixed, the ultrasonic vibrator 13 is firstly fixed at the bottom of the fixed base 15, and then the U-shaped clamping seat 23 and the alignment clamping block 25 are clamped and fixed.
When the ultrasonic vibrator 13 is fixed at the bottom of the fixed base 15, the limiting sleeve 1301 is directly aligned with the limiting column 16 and sleeved, then the ultrasonic vibrator 13 is pushed to be propped against the fixed base 15 for clamping, when the ultrasonic vibrator 13 moves, the ultrasonic vibrator 13 can prop against the inclined surface of the wedge-shaped clamping fixed block 21, after the inclined surface of the wedge-shaped clamping fixed block 21 is pressed, the ultrasonic vibrator can move towards the inner cavity of the hollow square column 18 and drive the square positioning slide block 19 to move, the square positioning slide block 19 is synchronously enabled to squeeze the first pressure spring 20, when the ultrasonic vibrator 13 continues to move, the ultrasonic vibrator is finally propped against the bottom of the fixed base 15, at the moment, the position of the wedge-shaped clamping fixed block 21 and the position of the clamping groove 1302 are propped against each other, and the first pressure spring 20 stretches at the moment and ejects the wedge-shaped clamping fixed block 21 into the clamping groove 1302, so that the ultrasonic vibrator 13 is fixed.
When the ultrasonic vibrator 13 needs to be disassembled, the handles of the two groups of square positioning slide blocks 19 are pulled first to drive the square positioning slide blocks 19 to move and enable the wedge-shaped clamping fixing blocks 21 to be separated from the clamping grooves 1302, and the ultrasonic vibrator 13 can be directly pulled out.
When the U-shaped clamping seat 23 and the alignment clamping block 25 are clamped and fastened, the U-shaped clamping seat 23 and the alignment clamping block 25 are abutted firstly, the alignment clamping block 25 is aligned to the bayonet of the U-shaped clamping seat 23 and clamped, then the pull rod 30 is pulled upwards, the pull rod 30 drives the sliding circular plate 27 to slide, the sliding circular plate 27 drives the second pressure spring 29 to compress and simultaneously drives the locking rod 28 to move upwards, at the moment, the U-shaped clamping piece 26 is aligned to the alignment clamping block 25 and inserted into the side circular hole of the U-shaped clamping seat 23, after the U-shaped clamping piece is completely inserted, the second pressure spring 29 pushes the sliding circular plate 27 to move under the elastic force of the second pressure spring 29, and the sliding circular plate 27 drives the locking rod 28 to be inserted into the surface circular hole of the U-shaped clamping piece 26, so that the position of the U-shaped clamping piece 26 is fixed.
In the subsequent process, the clamping and fixing of the U-shaped clamping seat 23 and the alignment clamping block 25 are required to be released, and the operation opposite to the installation and fixing is required, the pull rod 30 is lifted upwards, the U-shaped clamping piece 26 is removed, and then the U-shaped clamping seat 23 and the alignment clamping block 25 can be separated.
When the device is used, because the service life of the ultrasonic vibrator 13 is relatively short, the device belongs to a vulnerable part, therefore, in the later maintenance, the structural design in the device is utilized, the ultrasonic vibrator 13 is conveniently fixed below the outdoor flowing shell 1 and is convenient to fix with the vibrating tube 12, the ultrasonic vibrator 13 is convenient to disassemble and assemble in the later period, the maintenance is convenient, the normal work of the vibrating tube 12 is ensured, and therefore, the reinforced fixed nozzle tube 10 can continuously break the mixed liquid into uniform liquid drops when the mixed liquid is sprayed, the treatment of cells to be detected is ensured to be good, the stability of the device is ensured, and the detection effect is ensured.
The foregoing has shown and described the basic principles and main features of the present invention and the advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (6)
1. A flow cytometry detection fluid path system comprising:
The device comprises a flow outdoor shell (1), wherein a reinforced fixed nozzle pipe (10) is arranged at the bottom of the flow outdoor shell (1), a vibrating pipe (12) is connected inside the reinforced fixed nozzle pipe (10) in a sliding mode, an ultrasonic vibrator (13) is arranged at the bottom of the flow outdoor shell (1), a vibrating head (14) is arranged at the bottom of the ultrasonic vibrator (13), and the vibrating head (14) is fixedly connected with the vibrating pipe (12);
The cell suspension conveying pipe (2), the cell suspension conveying pipe (2) is fixedly connected to the top of the outdoor flowing casing (1), the outer wall of the cell suspension conveying pipe (2) is fixedly connected with the mixing pipe (4), and the mixing pipe (4) is arranged in the inner cavity of the outdoor flowing casing (1);
The sheath liquid conveying pipe (3), the sheath liquid conveying pipe (3) is fixedly connected to the side face of the outdoor flowing casing (1), the sheath liquid conveying pipe (3) penetrates through the outdoor flowing casing (1) and is fixedly connected with the outer wall of the mixing pipe (4), the sheath liquid conveying pipe (3) is fixedly connected with a sheath liquid quality detection box body (5), and the sheath liquid quality detection box body (5) is provided with a cleanliness sensor (6), a PH value sensor (7), an absorbance sensor (8) and an osmotic pressure sensor (9);
The upper end of the mixing tube (4) is fixedly connected with a plurality of groups of array distributed fixing strips (401), and the array distributed fixing strips (401) are fixedly connected to the outer wall of the cell suspension conveying tube (2);
The bottom of the outdoor flowing casing (1) is fixedly connected with a fixed base (15), the ultrasonic vibrator (13) is fixedly connected below the fixed base (15), the bottom of the fixed base (15) is fixedly connected with a limit column (16), two sides of the ultrasonic vibrator (13) are fixedly connected with limit sleeves (1301), and the limit sleeves (1301) are movably sleeved on the limit column (16);
Clamping grooves (1302) are formed in two sides of the ultrasonic vibrator (13), vertical plates (17) are fixedly connected to the bottom of the fixing base (15), hollow square columns (18) are fixedly connected to the side walls of the vertical plates (17), square positioning sliding blocks (19) are slidably connected to the inside of the hollow square columns (18), first pressure springs (20) are fixedly connected to one sides of the square positioning sliding blocks (19), wedge-shaped clamping fixing blocks (21) are fixedly connected to the other sides of the square positioning sliding blocks (19), and the wedge-shaped clamping fixing blocks (21) penetrate through the hollow square columns (18) and the vertical plates (17) and are movably clamped in the clamping grooves (1302).
2. A flow cytometry detection fluid pathway system as described in claim 1 wherein: the bottom of square location slider (19) is fixedly connected with handle, the fluting that corresponds with square location slider (19) handle is seted up to the bottom of cavity square column (18).
3. A flow cytometry detection fluid pathway system as described in claim 1 wherein: the side wall fixedly connected with second connecting block (24) of vibrations head (14), the lateral wall fixedly connected with chucking piece (25) of counterpoint of second connecting block (24), the lateral wall fixedly connected with first connecting block (22) of vibration pipe (12), the lateral wall fixedly connected with U-shaped cassette (23) of first connecting block (22), U-shaped cassette (23) and counterpoint chucking piece (25) joint, spout (2201) corresponding with first connecting block (22) have been seted up to the lateral wall of strenghthened type fixed nozzle pipe (10).
4. A flow cytometry detection fluid pathway system as described in claim 3 wherein: the side walls of the U-shaped clamping seat (23) and the alignment clamping block (25) are movably inserted with a U-shaped clamping piece (26), and two groups of round holes corresponding to the U-shaped clamping piece (26) are formed in the side walls of the U-shaped clamping seat (23) and the alignment clamping block (25).
5. The flow cytometry detection fluid pathway system of claim 4, wherein: the inside of counterpoint chucking piece (25) is equipped with the cylinder groove, sliding connection has slip plectane (27) in the cylinder groove of counterpoint chucking piece (25), the bottom fixedly connected with locking lever (28) of slip plectane (27), locking lever (28) pass counterpoint chucking piece (25) and the activity peg graft in U-shaped chucking piece (26), set up the round hole that corresponds with locking lever (28) on U-shaped chucking piece (26), the top fixedly connected with second compression spring (29) and pull rod (30) of slip plectane (27), the top of pull rod (30) passes counterpoint chucking piece (25) and upwards extends.
6. A flow cytometric assay method of a flow cytometric assay fluid pathway system according to any of claims 1-5, characterized in that:
The detection method comprises the following steps:
step S1: preparing cells or particles to be tested into a cell suspension, and then pressing a sample to be tested into a cell suspension conveying pipe (2) under a certain pressure;
Step S2: the sheath liquid is conveyed by a cell suspension conveying pipe (2), the sheath liquid is detected when passing through a sheath liquid quality detection box body (5), and the sheath liquid is directly pressurized and conveyed into the interior of a mixing pipe (4) when the cleanliness, the PH value, the absorbance and the osmotic pressure of the sheath liquid are normal;
step S3: when the cell suspension moves downwards, the cell suspension is finally mixed with the sheath liquid to form mixed liquid, the sheath liquid is wrapped outside the cell suspension for protection under the action of pressure, and the mixed liquid moves downwards and passes through the inner cavity of the flow chamber outer shell (1) to be sprayed out by the reinforced fixed nozzle pipe (10);
step S4: when passing through the reinforced fixed nozzle pipe (10), the vibrating pipe (12) is continuously vibrated by the ultrasonic vibrator (13) to move up and down, the mixed liquid is broken into uniform liquid drops, cells to be detected are dispersed in the liquid drops, and finally the liquid drops are injected into the laser detection pipeline (11);
Step S5: when the fluorescent substance passes through the laser detection pipeline (11), a laser source is arranged on the side surface of the laser detection pipeline (11), the focused light beam vertically irradiates on the mixed liquid, when the cell carries the fluorescent substance to pass through a laser irradiation area, fluorescent signals representing different substances and different wavelengths in the cell are generated, and the signals are emitted to a spatial omnibearing solid angle by taking the cell as the center to generate scattered light and fluorescent signals;
Step S6: the fluorescence generated by the fluorescence-dyed cells after being excited by the appropriate light waves is converted into an electric signal through a photoelectric converter, and the output electric signal is amplified and then transmitted to a computer, and finally data processing and analysis are carried out, so that the detection of the cells is formed.
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| US6003678A (en) * | 1997-08-21 | 1999-12-21 | University Of Washington | Particle separating apparatus and method |
| CN101290313B (en) * | 2007-04-16 | 2013-04-17 | 深圳迈瑞生物医疗电子股份有限公司 | Stream type cell device and method |
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