CN106840817A - A kind of Full-automatic circulation positive tumor cell enriching apparatus - Google Patents
A kind of Full-automatic circulation positive tumor cell enriching apparatus Download PDFInfo
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Classifications
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- G—PHYSICS
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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
- G01N1/40—Concentrating samples
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Abstract
The invention discloses a kind of Full-automatic circulation positive tumor cell enriching apparatus, including magnetic enrichment unit, centrifugal unit, buanch unit, dye unit and storing unit；The magnetic enrichment unit is used to be enriched with the circulating tumor cell for being connected with magnetic-particle, the centrifugal unit is used for the centrifugation of liquid, the buanch unit is used for the transfer of sample, reagent and material, and the dye unit is used to dye target cell, and the storing unit is used for the storage of reagent and material.The present invention is using magnetic immuno capture principle enrichment CTC, centrifugation of the invention is easy to cooperative mechanical arm to work using step-by-step movement movement sample channel position, mechanical arm is coordinated by shortest path with sample channel, mechanical arm can be saved and move repeatedly required time, accelerate the progress of whole flow process；The deceleration curve of centrifuge using intelligence it is controllable have brake deceleration, compared with traditional centrifuge free parking, that is, avoid disturbance of the centrifugation parking to liquid, centrifugation time has been saved again.
Description
Technical field
It is thin the present invention relates to circulating tumor cell fully-automated synthesis apparatus field, more particularly to a kind of Full-automatic circulation tumour
Born of the same parents' positive enriching apparatus.
Background technology
Circulating tumor cell (Circulating Tumor Cell, CTC) is that all kinds of tumours being present in peripheral blood are thin
The general designation of born of the same parents, it can be spontaneous or be come off from entity tumor focus (primary tumor or MET) by operation of diagnosis and treatment, into body circulation
System (such as peripheral blood).Most of CTC can occur apoptosis or swallowed after entering peripheral blood, and minority can escape and anchor focus
On tissue in addition or organ, and then develop into MET, increase malignant tumor patient mortality risk.Therefore, CTC detections can
It is effectively applied to external early diagnosis, prognostic evaluation, (including rapid clinical chemotherapeutics is screened and drug resistance individualized treatment
Detection), tumor recurrence monitoring and the exploitation of new tumour medicine etc..
Conventional CTC detections are divided into two steps, i.e. CTC enrichments (or capture) step and CTC detection and analysis steps.It is conventional
CTC capture techniques have Flow Cytometry, microfluidic chip technology and immunomagnetic bead technique.Wherein, immunomagnetic bead technique is fitted
Conjunction is separated under the conditions of large volume loading to a small amount of special cells, especially advantageous for CTC detection applications, its operation step
It is rapid relatively easy, it is easy to accomplish automation.
At present, in the market has occurred CTC enrichments and product or the service of detection, need manually operated mostly；It is existing
Automation equipment is generally import equipment and controlling organization is more, system running efficiency (time-consuming) and flux are relatively low.For example,
The CellSearch systems of Johson ＆ Johnson are managed based on immunological magnetic bead sorting primary, only realize the partial automation of detection operation, its
In cell enrichment (or capture) operation need to carry out by hand, automaticity is low, causes the efficiency of whole CTC detections program low
Under.Specifically, there is following deficiency in the CellSearch systems of Johson ＆ Johnson：(1) reagent cartridge is mixed, it is necessary to clean pipe
, there is cross contamination risk on road；(2), it is necessary to transfer CTC repeatedly in enrichment process；(3) hatching process is static；(4) need
Biological specimen is pre-processed；(5) flux is low.
The content of the invention
The present invention is intended to provide a kind of Full-automatic circulation positive tumor cell enriching apparatus, including magnetic enrichment unit, centrifugation
Unit, buanch unit, dye unit and storing unit；The magnetic enrichment unit is used to be enriched with the circulating tumor for being connected with magnetic-particle
Cell, the centrifugal unit is used for the centrifugation of liquid, and the buanch unit is used for the transfer of sample, reagent and material, the dye
Color element is used to dye target cell, and the storing unit is used for the storage of reagent and material.
Further, the centrifugal unit and the dye unit are arranged at the both sides on described device operating surface respectively,
The buanch unit is arranged at the top of described device operating surface, its can centrifugal unit, dye unit and storing unit it
Between transport sample, reagent and material, the magnetic enrichment unit and the storing unit are arranged on device operating surface suitable position
Put, so that the structure of the device is as compact as possible.
Preferably, the sample of present invention treatment is blood sample.
Further, the magnetic enrichment unit includes multiple bar magnets or magnetic patch, can be electromagnet or permanent magnet；The bar magnet
For directly being adsorbed in sample liquid and the CTC for being connected with magnetic-particle is enriched with, the magnetic patch is arranged at outside sample liquid, and
Magnetic field is produced in sample liquid, the CTC for being connected with magnetic-particle is enriched at one.Preferably, the present invention uses magnetic immuno
Capture agent, such as magnetic bead coupled antibody, carry out specific recognition and with reference to circulating tumor cell surface antigen so as to realize catching for CTC
Obtain and be enriched with.
In the preferred embodiment of the present invention, the magnetic enrichment unit includes multiple bands whether there is the magnetic of magnetic overcoat
Rod, each bar magnet one sample of correspondence, when rich magnetic particle in bar magnet insertion sample liquid, this is placed on described without magnetic overcoat
The outside of bar magnet, magnetic-particle can be enriched in the surface without magnetic overcoat, when the magnetic-particle of enrichment needs to be released
Bar magnet need to only be extracted out the effect for being capable of achieving degaussing from overcoat.Preferably, the bar magnet only has the bottom surface to have magnetic, so
Captured CTC can be made to be enriched in the bottom surface of bar magnet.Preferably, the material without magnetic overcoat is plastics.
Further, the centrifuge that the centrifugal unit is used is step-by-step movement centrifuge, and sample can be arbitrarily moved as needed
Channel position；Further, the step-by-step movement movement sample channel position is：Placed on centrifuge during machine on operator from
When heart pipe or buanch unit addition and absorption liquid, centrifuge can be moved slowly clockwise or counterclockwise, will have been operated
Passage is removed, and adjacency channel is gone into operative position.Using step-by-step movement movement sample channel position, it is easy to upper machine to place sample
This pipe, while buanch unit can be made to coordinate by shortest path with sample channel, Neng Goujie in order to coordinate buanch unit to work
About buanch unit moves repeatedly the time that position is consumed, and accelerates the progress of whole flow process.
Preferably, the centrifugal unit can in real time remember and record the position of each sample channel, be easy at the beginning of sample
The correspondence of the numbering that begins and extract/analyte sample numbering, and optional position of each biological specimen in centrifugal process
Having path memory and dynamic can set and control with static position.
It is highly preferred that the centrifuge is digitlization centrifuge, in centrifugal process, operator can be according to the centrifugation of sample
It is required that setting accelerates and deceleration curve, the deceleration curve of use be intelligence it is controllable have brake deceleration, do not disturbing sample layering
On the basis of, can Quick brake, save the time, and existing centrifuge slows down, in order to avoid the layering of sample is disturbed, all
It is to take without brake deceleration, deceleration time is very long.
Further, centrifugal unit of the present invention also has mixing mechanism, and the mixing mechanism is arranged on centrifuge
Each sample channel bottom, the mechanism can drive centrifuge tube to be spinned and/or jigging motion.
It is further preferred that the centrifugal unit also include visual identifying system, for centrifugal liquid is layered imaging,
Each layer draws the calculating of amount of liquid and height.
Preferably, the computational methods of the visual identifying system are divided into two parts, are (upper and lower according to visual identity photo
Layer is distinguish between with photoelectric sensor to color) come what is calculated：
1) the supernatant liquid height value that need to be taken out is calculated：Height subtracts photo and connects with lower floor at the middle and upper levels according to where sample
The difference of the height at place, adds the height (preventing the excessively multipair lower floor of drawing liquid from causing disturbance) of adjustment；
2) the liquid height value that need to be added is calculated：With the final height for highly subtracting existing liquid level of setting, go back if necessary
The height of the low dose of reagent of required addition before subtracting addition target liq.
Further, the buanch unit is made up of one or more mechanical arms, the mechanical arm can carry out up and down, left and right
And/or pivot；Preferably, the mobile system of the compatible hamilton of the mechanical arm；In a kind of preferred reality of the invention
Apply in mode, the multiple mechanical arm is respectively used to transfer sample, waste liquid, reagent and solid material；Of the invention another
In kind of preferred embodiment, the characteristics of the mechanical arm has " arm is multiplex ", can be used to pipette liquid or solid material.
In the preferred embodiment of the present invention, the Full-automatic circulation positive tumor cell enriching apparatus also include
Stirring rod.Preferably, the bar magnet and/or the stirring rod are captured by the mechanical arm by electromagnetic field and realize transfer.
It is further preferred that the stirring rod is propeller-like.
Preferably, the mechanical arm can drive the stirring rod or the bar magnet to carry out spinning motion, so as to realize stirring
With the action for mixing.
Further, the dye unit includes dyeing platform and cleaning platform, and the dyeing platform provides immunofluorescence dye
Color reagent, for being dyeed for CTC, the cleaning platform provides cleaning fluid, for cleaning impurity and unnecessary coloring agent.
Preferably, when the magnetic enrichment unit is bar magnet, the dye unit includes dyeing platform, cleaning platform and magnetic
Rod support frame, the dyeing platform and the cleaning platform are the rack for test tube equipped with dyeing liquor and cleaning fluid, when dyeing or cleaning
Only need to be by bar magnet enrichment positions insertion test tube.
Further, the storing unit includes reagent storage bin, consumptive material storage bin, bar magnet storage bin and discarded object storing
Storehouse, further may also include stirring rod storage bin.
Preferably, the reagent includes that circulating tumor cell immunocapture reagent, immunofluorescence dyeing reagent, cell are fixed
Reagent, buffer solution；Wherein, the circulating tumor cell immunocapture reagent contain can specific recognition and combine circulating tumor it is thin
The magnetic bead coupled antibody of cellular surface antigen；The stain for cell is preferably can specific recognition and combination circulating tumor cell table
The fluorescence antibody of face antigen.Further, the reagent can also be including in nucleic acid staining agent, capture Contrast agent, penetrating reagent
One or more.
Preferably, the consumptive material includes disposable liquid transfer gun head.
In the preferred embodiment of the present invention, the dye unit by the rack for test tube that can move up and down and
Bar magnet support frame is constituted, and dyeing test tube and cleaning test tube are placed with the rack for test tube, when the CTC being enriched with bar magnet needs to be cleaned
When, cleaning test tube need to be only moved to below corresponding bar magnet and bar magnet is immersed in cleaning fluid, when what is be enriched with bar magnet
When CTC needs to be colored, dyeing test tube need to be only moved to below corresponding bar magnet and bar magnet is immersed in dyeing liquor.Can
Mobile can be not only rack for test tube, can also be in another embodiment that bar magnet support frame may move.
Further, Full-automatic circulation positive tumor cell enriching apparatus of the present invention also include collecting and surveying unit, magnetic
Property the circulating tumor cell that obtains of enrichment be collected in this element, and can further be counted or be analyzed.
Preferably, the collection and analysis unit also has adsoption-magnetic field, when the CTC that bar magnet carries enrichment inserts collection and analysis
After in unit, bar magnet is extracted out from without magnetic overcoat, while opening the adsoption-magnetic field for collecting and surveying unit, so connection is magnetic
The magnetic field generation face that the CTC of particle is more easy to be collected analytic unit is adsorbed, and is laid on the face, is conducive to follow-up cell
Count and analysis work.
Further, centrifugal unit of the present invention also includes outer magnetic system, and the outer magnetic system includes permanent magnet or electricity
Magnet, for producing externally-applied magnetic field to the sample liquid in centrifuge tube in centrifuge, when immunocapture reagent is added into sample
Immunocapture reagent can be just promoted to be completely combined with CTC, it is necessary to be incubated a period of time when middle, can be by outer during incubation
Plus capture of the reinforced immunological magnetic capture agent to CTC is carried out in magnetic field, immune magnetic is such as controlled to catch by changing the direction of externally-applied magnetic field
The motion that reagent is oriented is obtained, so as to more fully be combined from the CTC in all directions in sample.
In the preferred embodiment of the present invention, the outer magnetic system is provided in different directions around centrifuge tube
Polylith magnet, these magnets can produce periodically variable magnetic field to strengthen capture effect.Of the invention another preferred
In implementation method, the outer magnetic system is provided in the magnet of specific location on centrifuge circumference, by low speed rotary centrifugal
Machine makes the distance between centrifuge tube and described outer magnetic system change, so as to produce periodically variable magnetic to centrifuge tube
, invisible spectro immunocapture reagent can carry out the directed movement of all directions under the influence of a magnetic field, so that immunocapture
Reagent is preferably combined with CTC.
Additionally, the present invention also provides a kind of Full-automatic circulation positive tumor cell enrichment method, comprise the following steps：
1) in centrifugal unit, after whole blood sample is centrifuged, carries out visual identity and calculate；Abandoning supernatant, adds slow
Fliud flushing, mixes, and adds immunocapture reagent, is incubated under the effect of outer magnetic system, then captures circulating tumor cell with bar magnet；
2) in dye unit, bar magnet transfer and cleaning are carried out, adds immunofluorescence dyeing reagent to carry out stain incubation, then
Cleaned；
3) in unit is collected and surveyed, bar magnet transfer is carried out, is demagnetized, be circulated the magnetic transfer of tumour cell, and carry out
Count or analyze.
Preferably, aforesaid operations are completed by mechanical arm.
The invention has the advantages that：
(1) (the immune magnetic capture and immune similar to CellSearch systems of device of the invention is used capture principle
Fluorescent staining), but compared with circulating tumor cell positive enrichment system (for example, CellSearch) for having listed, its main spy
Point has been compatible Sample pretreatment process and (blood sample has been diluted, and CellSearch systems are needed before upper machine manually
Coordinate desk centrifuge that treatment is diluted to sample).
(2) centrifugal unit of the invention is easy to cooperative mechanical arm to work using step-by-step movement movement sample channel position, makes machine
Tool arm is coordinated with sample channel by shortest path, can be saved mechanical arm and be moved repeatedly required time, accelerates whole flow process
Progress；The deceleration curve of centrifuge used by the centrifugal platform using intelligence it is controllable have brake deceleration, with traditional centrifuge
Free parking is compared, that is, avoid disturbance of the centrifugation parking to liquid, and centrifugation time has been saved again.
(3) compared with CellSearch, the advantage of device or method of the present invention has：A. the sample of each passage and examination
Agent operation is independent, and sample and reagent do not have cross pollution, and CellSearch is to clean pipeline the need for intersecting；
B.CellSearch is in enrichment process, it is necessary to transfer CTC repeatedly, the present invention is once to capture in enrichment process, not
Need transfer；C. hatching process, CellSearch is static, and the present invention is dynamic, has stirring, mixes and outer magnetic strength；
D.CellSearch needs to pre-process biological specimen, and the present invention is full automatic；E. passage of the invention is more, higher
Effect.
Brief description of the drawings
Fig. 1 is the structural representation of a preferred embodiment of the invention；
Fig. 2 is the structural representation of another preferred embodiment of mechanical arm in the present invention；
Fig. 3 is a structural representation for preferred embodiment of bar magnet in the present invention.
Specific embodiment
The present invention is described in further detail with embodiment below in conjunction with the accompanying drawings, but those skilled in the art are according to following
The replacement of concrete scheme, conversion or combination are both contained within protection scope of the present invention in embodiment.Filled using with the present invention
The liquid working station form of similar designs flow sample process is put, or uses the enrichment method similar with the inventive method,
Within concept of the invention.
Embodiment 1
Present embodiments provide a kind of Full-automatic circulation positive tumor cell enriching apparatus (Fig. 1), including magnetic enrichment unit,
Centrifugal unit, buanch unit, dye unit and storing unit；Wherein magnetic enrichment unit is made up of 12 bar magnets 1；Centrifugal unit
Critical piece is can be while process 12 digitlization centrifuges 2 of sample；Buanch unit is by with transverse direction (X) and longitudinal direction (Y) rail
Two of road mechanical head 3-1,3-2 compositions of transfer, this two mechanical heads of transfer can also on the direction in vertical operation face (Z, in figure
It is not shown) it is mobile；Dye unit include with sliding rail rack for test tube 4 and can vertical operation face movement bar magnet support frame 5,
Test tube equipped with staining reagent or cleaning fluid is set on demand in the rack for test tube；Storing unit includes reagent storage bin 6, liquid-transfering gun
Head storage bin 7, waste liquid storage bin 8, bar magnet storage bin 9 and stirring rod storage bin 10.Additionally, the present embodiment also includes collecting and surveying
Unit, the collection and analysis unit includes to be collected 12 samples the reading box 11 of analysis, in the top of the reading box
It is provided with bar magnet support frame 12.
Centrifugal unit and dye unit are separately positioned on the present embodiment the left and right sides of operation planar, are set therebetween
Storing unit, buanch unit is arranged on the top of operating surface, and its moving range can cover whole operation face, and this design can be excellent
The transfer distance of compound material, shortens run time.
Further, the centrifuge 2 of centrifugal platform is step-by-step movement centrifuge, and the passage position of sample can be arbitrarily moved as needed
Put, in a preferred embodiment, a as shown in Figure 1 and b is the operative position of mechanical arm, and pipetting is needed every time
When, sample centrifuge tube is gone to a and b, two transfer machinery head 3-1,3-2 are added or liquid draw mobile so far position simultaneously
Body, after fixed operative position, because the mobile route of mechanical arm is shortened, and mechanical arm and centrifuge simultaneously participate in displacement, make total
Operational process shorten.In addition the digitlization centrifuge can accelerate and deceleration curve according to the centrifugation of sample requirement setting, use
Deceleration curve to be that intelligence is controllable have brake deceleration, on the basis of sample layering is not disturbed, can Quick brake, one can be entered
Step is time-consuming.The centrifuge can also in real time remember and record the position of each sample channel, be easy to sample initial number
The correspondence numbered with extract/analyte sample, and optional position Shi You road of each biological specimen in centrifugal process
Footpath is remembered, and dynamic can set and control with static position.
The mechanical arm of the buanch unit is connected with two transfers mechanical head 3-1 and 3-2, can insert and take and discharge liquid-transfering gun
Head, draw and blowout liquid, additionally can by electromagnetic field capture, transfer and rotation bar magnet and stirring rod, the bar magnet and
The upper end of stirring rod is both provided with ferromagnetic material, can be transferred mechanical head crawl.
In another embodiment of the invention, mechanical arm can also be arranged in addition to it can set rectilinear orbit
The form being pivoted, as shown in Fig. 2 the centrifugal unit has two rotary mechanical arms 3-3 and 3-4, their storages from left side
Pipette tips, reagent etc. are pipetted in feed bin 13, is that centrifuge 2 provides liquid transfer.
Bar magnet structure in the present embodiment is as shown in figure 3, in addition to upper end carries ferromagnetic material 11, its bottom surface 14 can
Release magnetic field, the magnetic field be by bar magnet inside magnetic core 12 discharge, outside it also have without magnetic overcoat 13, magnetic during work
Property particle can be enriched in the bottom surface without magnetic overcoat 13, only need to extraction be releasable magnetic from overcoat by bar magnet after the completion of enrichment
Property particle, in addition its top be additionally provided with screens part 15, for bar magnet to be suspended on into bar magnet support frame.
The centrifugal unit of the present embodiment also includes visual identifying system, and the imaging of centrifugal liquid layering, each layer can be inhaled
Take amount of liquid and height is calculated.
Dye unit in the present embodiment includes dyeing platform, cleaning platform and bar magnet support frame 5, and the dyeing platform is carried
For immunofluorescence dyeing reagent, the cleaning platform provides cleaning fluid, and they are merged to be arranged on and same can be slided along track
On rack for test tube 4, bar magnet support frame can vertical operation face move up and down, therefore staining reagent need to be only moved under bar magnet when dyeing
Side, bar magnet support frame is moved down by bar magnet is immersed in staining reagent, it is necessary to then move under bar magnet cleaning reagent when cleaning
Side, you can cleaned bar magnet immersion cleaning reagent.
The present embodiment also includes collecting and surveying unit, is made up of reading box 11 and bar magnet support frame 12, after it will be enriched with
Bar magnet insert in corresponding reading box, extract bar magnet out and only retain without magnetic overcoat, while open the adsoption-magnetic field in reading cassette bottom face,
The CTC for being connected with magnetic-particle can be adsorbed by bottom surface, and be laid on the face, be that follow-up cell count and analysis are provided with
The condition of profit.
Further, centrifugal unit described in the present embodiment also includes outer magnetic system (not shown), the outer magnetic system bag
Permanent magnet or electromagnet are included, for producing externally-applied magnetic field to the sample liquid in centrifuge tube in centrifuge, when immunocapture examination
Immunocapture reagent can be just promoted to be completely combined with CTC, it is necessary to be incubated a period of time when agent is added into sample, in the mistake of incubation
Captures of the Cheng Zhongke by externally-applied magnetic field come reinforced immunological magnetic capture agent to CTC, such as by change the direction of externally-applied magnetic field come
The motion that the immune magnetic capture agent of control is oriented, so as to more fully be combined from the CTC in all directions in sample.
In the preferred embodiment of the present invention, the outer magnetic system is provided in different directions around centrifuge tube
Polylith magnet, these magnets can produce periodically variable magnetic field to strengthen capture effect.Of the invention another preferred
In implementation method, the outer magnetic system is provided in the magnet of specific location on centrifuge circumference, by low speed rotary centrifugal
Machine makes the distance between centrifuge tube and described outer magnetic system change, so as to produce periodically variable magnetic to centrifuge tube
, invisible spectro immunocapture reagent can carry out the directed movement of all directions under the influence of a magnetic field, so that immunocapture
Reagent is preferably combined with CTC.
Full-automatic circulation positive tumor cell enriching apparatus in the present embodiment can simultaneously process 12 samples, and compatibility
Sample pretreatment process, run time is short (fully loaded whole process operation time-consuming about 2 hours 40 points), realizes the efficient place of sample
Reason.
Embodiment 2
A kind of circular of preferred embodiment of visual identifying system of the present invention includes two parts, respectively
It is the calculating for drawing liquid height and the calculating for needing adding liquid height, is all the photo according to captured by visual identifying system
Calculated.
The supernatant liquid height value that calculating need to be taken out：Subtracted according to height where sample photo at the middle and upper levels with lower floor joint
Height difference, add the height (prevent the excessively multipair lower floor of drawing liquid from causing disturbance) of adjustment；
The liquid height value that calculating need to be added：With the final height for highly subtracting existing liquid level of setting, also if necessary
The height of the low dose of reagent of required addition before subtracting addition target liq.
For example, acquiescence whole blood sample volume is 10mL, sample is divided into upper plasma and lower floor's haemocyte after centrifugation, in absorption
After layer blood plasma, the immunocapture reagent and capture Contrast agent of 0.2mL need to be added, then add dilution buffer to 5mL, wherein
The height of adjustment is the corresponding height of 0.8mL liquid, then total step that it is calculated is two steps：
1) height value of the upper plasma that need to be taken out is calculated：Height where 10mL samples subtract photo at the middle and upper levels blood plasma with
The difference of the height of layer haemocyte joint, adds the height (the corresponding height of 0.8mL liquid) of 7mm；
2) the dilution buffer height value that need to be added is calculated：Height value where required dilution buffer is 5mL subtracts extraction
The low dose of reagent sum of liquid level after blood plasma and the 0.2mL that need to be added before dilution buffer；
Example：The upper plasma of 7mL is had in 10mL samples, then decimation value is 6.2mL (7-0.8), the dilution that need to be added is delayed
Fliud flushing is 1mL (5-3.8-0.2).
Embodiment 3
A kind of positive enrichment method of Full-automatic circulation tumour cell, mainly includes the following steps that：
1) in centrifugal platform, after whole blood sample is centrifuged, carries out visual identity and calculate；Abandoning supernatant, adds buffering
Liquid, mixes, and adds immunocapture reagent, is incubated under the effect of outer magnetic system, then captures circulating tumor cell with bar magnet；
2) in dyeing platform, bar magnet transfer and cleaning are carried out, adds immunofluorescence dyeing reagent to carry out stain incubation, then enter
Row cleaning；
3) in unit is collected and surveyed, bar magnet transfer is carried out, is demagnetized, the magnetic transfer of tumour cell is circulated, with after an action of the bowels
It is continuous further it to be counted and is analyzed..
Aforesaid operations are completed by mechanical arm.
Specifically, the automation mechanized operation step of the positive enrichment method can be found in table 1.
The positive enrichment automation mechanized operation steps of the CTC of table 1
Preferred embodiment of the invention described in detail above.It should be appreciated that one of ordinary skill in the art without
Need creative work just can make many modifications and variations with design of the invention.Therefore, all technologies in the art
Personnel are available by logical analysis, reasoning, or a limited experiment on the basis of existing technology under this invention's idea
Technical scheme, all should be in the protection domain being defined in the patent claims.
Claims (14)
1. a kind of Full-automatic circulation positive tumor cell enriching apparatus, it is characterised in that including magnetic enrichment unit, centrifugal unit, turn
Move unit, dye unit and storing unit；The magnetic enrichment unit is used to be enriched with the circulating tumor cell for being connected with magnetic-particle, institute
Centrifugal unit is stated for the centrifugation of liquid, the buanch unit is used for the transfer of sample, reagent and material, the dye unit is used
Dyeed in target cell, the storing unit is used for the storage of reagent and material.
2. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the magnetic enrichment list
Unit includes multiple bar magnets or magnetic patch.
3. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the magnetic enrichment list
Unit includes multiple bands whether there is the bar magnet of magnetic overcoat.
4. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the centrifugal unit
The centrifuge for using is step-by-step movement centrifuge, and the channel position of sample can be arbitrarily moved as needed.
5. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 4, it is characterised in that operator can basis
The centrifugation of sample requires acceleration and the deceleration curve of the setting centrifuge.
6. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the centrifugal unit
Including visual identifying system, the calculating of imaging, each layer absorption amount of liquid and height for being layered to centrifugal liquid.
7. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the buanch unit
Be made up of one or more mechanical arms, the mechanical arm can carry out up and down, left and right and/or pivot.
8. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that also including stirring rod.
9. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the dye unit
Including dyeing platform and cleaning platform, the dyeing platform provides immunofluorescence dyeing reagent, described clear for being dyeed for CTC
Wash platform and cleaning fluid is provided, for cleaning impurity and unnecessary coloring agent.
10. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the storing unit
Including reagent storage bin, consumptive material storage bin, bar magnet storage bin and discarded object storage bin.
11. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that also including collection point
Analysis unit, the circulating tumor cell that magnetite gathering is obtained is collected in this element and is further counted or analyzed.
12. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 11, it is characterised in that the collection point
Analysis unit has adsoption-magnetic field.
13. Full-automatic circulation positive tumor cell enriching apparatus as claimed in claim 1, it is characterised in that the centrifugal unit
Also include outer magnetic system, for producing externally-applied magnetic field to the sample liquid in centrifuge tube in centrifuge.
14. a kind of Full-automatic circulation positive tumor cell enrichment methods, it is characterised in that comprise the following steps：
1) in centrifugal unit, after whole blood sample is centrifuged, carries out visual identity and calculate；Abandoning supernatant, adds buffer solution,
Mix, add immunocapture reagent, be incubated under the effect of outer magnetic system, then capture circulating tumor cell with bar magnet；
2) in dye unit, bar magnet transfer and cleaning are carried out, adds immunofluorescence dyeing reagent to carry out stain incubation, then carry out
Cleaning；
3) in unit is collected and surveyed, bar magnet transfer is carried out, is demagnetized, the magnetic transfer of tumour cell is circulated, subsequently to enter
One step is counted and analyzed to it.
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