CN206593934U - A kind of negative enriching apparatus of Full-automatic circulation tumour cell - Google Patents
A kind of negative enriching apparatus of Full-automatic circulation tumour cell Download PDFInfo
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- CN206593934U CN206593934U CN201720246751.0U CN201720246751U CN206593934U CN 206593934 U CN206593934 U CN 206593934U CN 201720246751 U CN201720246751 U CN 201720246751U CN 206593934 U CN206593934 U CN 206593934U
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Abstract
The utility model discloses a kind of negative enriching apparatus of Full-automatic circulation tumour cell, centrifugal unit, buanch unit, storing unit and collection and analysis unit；The centrifugal unit is used for the centrifugation of liquid and the dyeing of cell, cleaning and is incubated, the buanch unit is used for the transfer of sample, reagent and material, the storing unit is used for the storage of reagent and material, and the circulating tumor cell that the collection and analysis unit is used to collect enrichment acquisition is counted or analyzed.Centrifugation of the present utility model is easy to cooperative mechanical arm to work, mechanical arm is coordinated with sample channel by shortest path using step-by-step movement movement sample channel position, can be saved mechanical arm and be moved 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 saved again.
Description
Technical field
The utility model is related to circulating tumor cell fully-automated synthesis apparatus field, more particularly to a kind of Full-automatic circulation swells
Oncocyte feminine gender 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 come off by operation of diagnosis and treatment from entity tumor focus (primary tumor or transfer stove), 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 or organ in addition, and then develop into transfer stove, 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..
At present, in the market has occurred CTC enrichments and product or the service of detection, unique CTC detections for obtaining FDA approvals
Equipment is CellSearch systems, and the system is the detection device developed based on immuno magnetic cell separation technology, and it utilizes mark
Note has the magnetic for epithelial cell adhesion molecule (Epithelial Cell Adhesion Molecule, EpCAM) antibody
Grain carries out CTC capture, and this method has the advantages that high specificity, but its None- identified does not express the unknown thin of target label
Born of the same parents, and its capture rate or relatively low, the interference of leucocyte is larger.In addition, current some CTC detection devices and technology do not have still
There is the automatic control for realizing detection process, that is, be not carried out the automatic sampling of blood testing sample and reagent, cell
Automation capture and identification and analysis, therefore there is excessive human intervention in CTC detection so that CTC detection accuracy is low, inspection
Survey cost too high.
I.e. in view of a total of 15 partings of circulating tumor cell CTC, and CellSearch is only able to detect wherein one
Individual type, loss is very high, therefore, and those skilled in the art are directed to exploitation one kind and are screened with exclusive method principle and be enriched with CTC
Full-automatic device.
Utility model content
The utility model is intended to provide one kind negative enrichment cycles tumour cell from whole blood based on density-gradient centrifugation method
Method and its fully-automatic equipment, this method is layered using the density variation of variety classes cell, and is discarded with the precipitation method
Main interference cell, so as to obtain the circulating tumor cell for meeting certain density criterion.
To achieve the above object, the utility model provides a kind of negative enriching apparatus of Full-automatic circulation tumour cell, bag
Include centrifugal unit, buanch unit, storing unit and collect and survey unit；The centrifugal unit is used for centrifugation and the cell of liquid
Dyeing, cleaning and be incubated, the buanch unit be used for sample, reagent and material transfer, the storing unit be used for reagent
With the storage of material, the circulating tumor cell that the collection and analysis unit is used to collect enrichment acquisition is counted or analyzed.
Further, the centrifugal unit is using centrifuge as core, and the storing unit is arranged at the week of the centrifugal unit
Enclose, the buanch unit is arranged at the top of described device operating surface, it can be in the centrifugal unit and the storing unit
Between transhipment sample, reagent and material.
Preferably, the sample of the utility model processing is blood sample.
Further, the storing unit includes reagent storage bin, consumptive material storage bin and discarded object storage bin；Preferably, also
It may include stirring rod storage bin.
Further, density gradient centrifugation liquid, multi-joint antibody reagent, cell dyeing are at least stored in the reagent storage bin
Reagent and buffer solution.
Preferably, the reagent includes density gradient centrifugation liquid, multi-joint antibody reagent, immunofluorescence dyeing reagent, cell
Fixating reagent, buffer solution；Wherein, the multi-joint antibody reagent can combine at least two interference cells, be preferably that can combine blood
In leucocyte and red blood cell；The stain for cell is preferably can specific recognition and combination circulating tumor cell surface antigen
Fluorescence antibody.Further, the reagent can also include one kind in nucleic acid staining agent, capture Contrast agent, penetrating reagent
Or it is a variety of.
Maximum interference is leucocyte and red blood cell in negative concentration method, the above method using many len antibodies by leucocyte with
Erythrocyte binding gets up；In a kind of preferred embodiment of the present utility model, the density of the combination is larger, through density level bands
Degree is centrifuged and precipitated, and is located at as the CTC of target in intermediate layer, the layer in addition to target CTC, it is also possible to had and its density
Similar other cells, these other cells may be also CTC, or the worth non-CTC cells further analyzed and researched.
Preferably, the consumptive material includes disposable liquid transfer gun head.
Further, the centrifugal unit also include visual identifying system, for centrifugal liquid is layered be imaged, each layer inhale
Take the calculating of amount of liquid and height.
Preferably, the computational methods of the visual identifying system are divided into two parts, be according to visual identity photo (up and down
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 connected at the middle and upper levels with lower floor 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 added needed for before subtracting addition target liq.
Further, the centrifuge that the centrifugal unit is used can arbitrarily move sample as needed for step-by-step movement centrifuge
Channel position；Further, the step-by-step movement movement sample channel position is：Placed on operator during machine on centrifuge 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 the step-by-step movement mobile example channel position, it is easy to upper machine to place sample
QC, while in order to coordinate buanch unit to work buanch unit can be made to coordinate with sample product passage by shortest path, can
Save buanch unit and move repeatedly the time that position is consumed, accelerate the progress of whole flow process.
Preferably, the centrifugal unit can remember and record the position of each sample channel in real time, 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
There are path memory and dynamic to 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, and is disturbed in order to avoid the layering of sample, all
It is to take no brake deceleration, deceleration time is very long.
Further, centrifugal unit described in the utility model also has mixing mechanism, and the mixing mechanism is arranged on centrifugation
Each sample channel bottom of machine, the mechanism can drive centrifuge tube to be spinned and/or jigging motion.
Further, the buanch unit is made up of one or more mechanical arms, the mechanical arm can carry out above and below, left and right
And/or pivot；Preferably, the compatible hamilton of mechanical arm mobile system；It is excellent in one kind of the present utility model
Select in embodiment, the multiple mechanical arm is respectively used to transfer sample, waste liquid, reagent and solid material；It is new in this practicality
In another preferred embodiment of type, the characteristics of mechanical arm has " arm use ", available for pipetting liquid or solid material
Material.
In a kind of preferred embodiment of the present utility model, the negative enriching apparatus of the Full-automatic circulation tumour cell is also
Including stirring rod.Preferably, the stirring rod is captured by the mechanical arm by electromagnetic field and realizes transfer.
It is further preferred that the stirring rod is propeller-like.
Preferably, the mechanical arm can drive the stirring rod to carry out spinning motion, so as to realize that stirs and mix moves
Make.
Further, the collection and analysis unit has observation window, is available for operator to observe or machine automatic camera.
In addition, the utility model also provides a kind of Full-automatic circulation tumour cell negative enrichment method, comprise the following steps：
1) in centrifugal unit, be staggeredly placed the test tube A equipped with whole blood sample and the test tube equipped with density gradient centrifugation liquid
Clean test tube C is placed in B, the rack for test tube outside centrifugal unit；
2) buffer solution and cross-linking antibody reagent are added into whole blood sample test tube A, is mixed and is incubated；
3) whole blood sample after incubation is gently added in the test tube B that density gradient centrifugation liquid has been pre-installed on side, through centrifugation
Visual identity is carried out afterwards, and circulating tumor cell layer is drawn with calculating, then；
4) circulating tumor cell of absorption layer is moved into the test tube C outside centrifugal unit, and with buanch unit by test tube B from
Removed in centrifugal unit；
5) test tube C is moved into centrifugal unit with buanch unit, adds after buffer solution and centrifuge, supernatant discarding；
6) immunofluorescence dyeing reagent is added into test tube C, after dyed incubation, buffer solution is added, then abandon through centrifugation
After clear, add fixating reagent and mix；
7) circulating tumor cell after being resuspended with fixating reagent is transferred in collection and analysis unit and further divided
Analysis.
The utility model has the advantages that：
(1) negative enriching apparatus of the present utility model is that the automation for realizing negative enrichment cycles tumour cell first is set
Standby, compact conformation, the enrichment of circulating tumor cell and staining procedure can be completed in centrifugal unit, it is not necessary to which ad hoc dyeing is flat
Platform；
(2) centrifugal unit of the present utility model is easy to cooperative mechanical arm to work using step-by-step movement movement sample channel position,
Mechanical arm is coordinated with sample channel by shortest path, mechanical arm can be saved and move repeatedly required time, accelerate whole stream
The progress of journey；The deceleration curve of centrifuge used in the centrifugal platform using intelligence it is controllable have brake deceleration, with it is traditional from
Scheming free parking is compared, that is, avoids disturbance of the centrifugation parking to liquid, centrifugation time has been saved again.
(3) sample and reagent operation of each passage of the utility model are independent, and sample and reagent do not have cross pollution,
Incubation process of the present utility model is dynamic, there is stirring and outer magnetic strength, and the utility model needs to carry out biological specimen
The process of pretreatment is full automatic.
Brief description of the drawings
Fig. 1 is the structural representation of a preferred embodiment of the present utility model；
Fig. 2 is the structural representation of another preferred embodiment of mechanical arm in the utility model.
Embodiment
The utility model is described in further detail with embodiment below in conjunction with the accompanying drawings, but those skilled in the art according to
The replacement of concrete scheme, conversion or combination are both contained within protection domain of the present utility model in following examples.Using with
The liquid working station form of the utility model device similar designs flow sample process, or use and the utility model method class
As enrichment method, within concept of the present utility model.
Embodiment 1
Present embodiments provide a kind of negative enriching apparatus (Fig. 1) of Full-automatic circulation tumour cell, including centrifugal unit, turn
Move unit, storing unit and collect and survey unit；Wherein, the core component of centrifugal unit is the numeral with 12 sample channels
Change centrifuge 1, the centrifugal unit is used for the centrifugation of liquid and the dyeing of cell, cleaning and is incubated；Buanch unit by with
Laterally mechanical head 2-1, the 2-2 of (X) and longitudinal direction (Y) track two transfers is constituted, and this two mechanical heads of transfer can also grasped vertically
Make on the direction in face (Z, figure not shown in) mobile；Storing unit includes reagent storage bin 3, liquid transfer gun head storage bin 4, stirring rod
At least store density gradient centrifugation liquid in storage bin 5, test tube storage bin 6 and waste liquid storage bin 7, the reagent storage bin 3, it is many
Len antibody reagent, immunofluorescence dyeing reagent, cell fixating reagent and buffer solution, multi-joint antibody reagent can combine leucocyte with it is red
Cell；Unit is collected and surveyed, the collection and analysis unit includes multiple reading boxes 8, the circulating tumor for collecting enrichment acquisition is thin
Born of the same parents are counted or analyzed.
Further, the centrifuge 1 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 the operative position that b are mechanical arm, pipetting is needed every time
When, sample centrifuge tube is gone to a and b, mobile so far position is added or liquid draw two transfer machinery head 2-1,2-2 simultaneously
After body, 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 require that setting accelerates and deceleration curve according to the centrifugation of sample, 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
Walk the saving time.The centrifuge can also remember and record the position of each sample channel in real time, be easy to sample initial number
The correspondence numbered with extract/analyte sample, and optional position of each biological specimen in centrifugal process is that have road
Footpath is remembered, and dynamic can set and control with static position.
The centrifugal unit of the present embodiment also include visual identifying system, can to centrifugal liquid be layered be imaged, each layer inhale
Amount of liquid and height is taken to be calculated；In addition the channel bottom of centrifuge is additionally provided with mixing mechanism, can be by spinning and/or shaking
Rolling makes the liquid blending in sample.
The mechanical arm of the buanch unit is connected with two transfers mechanical head 2-1 and 2-2, can insert and take and discharge liquid-transfering gun
Head, draws and blowout liquid, can additionally be captured by electromagnetic field, shift stirring rod, the upper end of the stirring rod is provided with
Ferromagnetic material, can be transferred mechanical head crawl, Grasp Modes are not unique herein, and it is all feasible that can meet desired Grasp Modes
's.
In another embodiment of the present utility model, mechanical arm can also be set in addition to it can set rectilinear orbit
The form being pivoted is set to, as shown in Fig. 2 the centrifugal unit has two rotary mechanical arms 2-3 and 2-4, they are from left side
Pipette tips, reagent etc. are pipetted in the storage bin 9 that can be slided along track, is that centrifuge 1 provides liquid transfer.
As shown in fig. 1, in the centrifuge in the present embodiment sample channel be two one group, totally 6 groups, machine operation before
Whole blood sample is placed in a positions in every group of passage, and density gradient centrifugation liquid is placed in b positions, and buanch unit then can be in centrifuge a
Transfer liquid between passage and b passages, and transfer liquid and centrifuge tube between centrifuge and rack for test tube.
In a kind of preferred embodiment of the present utility model, the dilution step and density gradient centrifugation step of sample exist respectively
A the and b positions of centrifuge are carried out, and are collected the CTC cells obtained and are transferred in the clean tube on test tube storage bin 6, then
The test tube is transferred in centrifuge and carries out follow-up staining procedure, in this embodiment the transferable test tube of buanch unit, is shifted
The mode of test tube can be clamping type, magnetic gripping-type etc.；In another preferred embodiment of the present utility model, the dilution of sample
In test tube of the step on test tube storage bin 6 carry out, mixing is stirred with stirring rod, the liquid sample after mixing be transferred to from
A positions in scheming, density gradient centrifugation step is carried out in b positions, and test tube need not be shifted in this embodiment.
Device in the present embodiment can be with 6 samples of batch enrichment and detection, and compatible Sample pretreatment process, during operation
Between short, efficiency high.
Embodiment 2
A kind of circular of preferred embodiment of visual identifying system described in the utility model includes two parts,
It is to draw the calculating of liquid height and need the calculating of adding liquid height respectively, is all according to captured by visual identifying system
Photo is calculated.
Calculate the supernatant liquid height value that 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；
Calculate the liquid height value that 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 added needed for 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, absorption after centrifugation
After layer blood plasma, 0.2mL reagent need to be added, dilution buffer is then added to 5mL, wherein the height adjusted is 0.8mL liquid
Corresponding height, then its total step calculated is two steps：
1) height value for 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 7mm height (the corresponding height of 0.8mL liquid)；
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 and the 0.2mL that need to be added before dilution buffer after blood plasma；
Example：7mL upper plasma 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 negative enrichment method of Full-automatic circulation tumour cell, is mainly included the following steps that：
1) in centrifugal unit, be staggeredly placed the test tube A equipped with whole blood sample and the test tube equipped with density gradient centrifugation liquid
Clean test tube C is placed in B, the rack for test tube outside centrifugal unit；
2) buffer solution and cross-linking antibody reagent are added into whole blood sample test tube A, is mixed and is incubated；
3) whole blood sample after incubation is gently added in the test tube B that density gradient centrifugation liquid has been pre-installed on side, through centrifugation
Visual identity is carried out afterwards, and circulating tumor cell layer is drawn with calculating, then；
4) circulating tumor cell of absorption layer is moved into the test tube C outside centrifugal unit, and with buanch unit by test tube B from
Removed in centrifugal unit；
5) test tube C is moved into centrifugal unit with buanch unit, adds after buffer solution and centrifuge, supernatant discarding；
6) immunofluorescence dyeing reagent is added into test tube C, after dyed incubation, buffer solution is added, then abandon through centrifugation
After clear, add fixating reagent and mix；
7) circulating tumor cell after being resuspended with fixating reagent is transferred in collection and analysis unit and further divided
Analysis.
Specifically, the automation mechanized operation step of the negative enrichment method can be found in table 1.
Table 1CTC feminine gender enrichment automation mechanized operation steps
Preferred embodiment of the present utility model described in detail above.It should be appreciated that the ordinary skill people of this area
Member just can make many modifications and variations without creative work according to design of the present utility model.Therefore, all this technology necks
Technical staff passes through logic analysis, reasoning or limited reality on the basis of existing technology according to design of the present utility model in domain
Available technical scheme is tested, all should be in the protection domain being defined in the patent claims.
Claims (9)
1. the negative enriching apparatus of a kind of Full-automatic circulation tumour cell, it is characterised in that including centrifugal unit, buanch unit, storing
Unit and collection and analysis unit；The centrifugal unit is used for the centrifugation of liquid and the dyeing of cell, cleaning and is incubated, described turn
Moving unit is used for the transfer of sample, reagent and material, and the storing unit is used for the storage of reagent and material, the collection and analysis
The circulating tumor cell that unit is used to collect enrichment acquisition is counted or analyzed.
2. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 1, it is characterised in that the storing unit
Including reagent storage bin, consumptive material storage bin and discarded object storage bin.
3. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 2, it is characterised in that in reagent storage bin
At least store density gradient centrifugation liquid, multi-joint antibody reagent, cell staining reagent and buffer solution.
4. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 1, it is characterised in that the centrifugal unit
The centrifuge used can arbitrarily move the channel position of sample as needed for step-by-step movement centrifuge.
5. the negative enriching apparatus of Full-automatic circulation tumour cell 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. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 1, it is characterised in that the centrifugal unit
Including visual identifying system, for centrifugal liquid is layered be imaged, each layer draws the calculating of amount of liquid and height.
7. the negative enriching apparatus of Full-automatic circulation tumour cell 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 above and below, left and right and/or pivot.
8. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 1, it is characterised in that also including stirring rod.
9. the negative enriching apparatus of Full-automatic circulation tumour cell as claimed in claim 1, it is characterised in that the centrifugal unit
Also include mixing mechanism.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201720246751.0U CN206593934U (en) | 2017-03-14 | 2017-03-14 | A kind of negative enriching apparatus of Full-automatic circulation tumour cell |
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| CN201720246751.0U CN206593934U (en) | 2017-03-14 | 2017-03-14 | A kind of negative enriching apparatus of Full-automatic circulation tumour cell |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106840816A (en) * | 2017-03-14 | 2017-06-13 | 骏实生物科技(上海)有限公司 | A kind of Full-automatic circulation tumour cell feminine gender enriching apparatus |
| CN111760607A (en) * | 2020-06-08 | 2020-10-13 | 六盘水市第二人民医院 | A sample storage device for clinical laboratory |
| CN112899232A (en) * | 2021-03-03 | 2021-06-04 | 河南省肿瘤医院 | Automatic system for separating circulating tumor cells from blood sample |
-
2017
- 2017-03-14 CN CN201720246751.0U patent/CN206593934U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106840816A (en) * | 2017-03-14 | 2017-06-13 | 骏实生物科技(上海)有限公司 | A kind of Full-automatic circulation tumour cell feminine gender enriching apparatus |
| CN111760607A (en) * | 2020-06-08 | 2020-10-13 | 六盘水市第二人民医院 | A sample storage device for clinical laboratory |
| CN112899232A (en) * | 2021-03-03 | 2021-06-04 | 河南省肿瘤医院 | Automatic system for separating circulating tumor cells from blood sample |
| CN112899232B (en) * | 2021-03-03 | 2022-02-01 | 河南省肿瘤医院 | Automatic system for separating circulating tumor cells from blood sample |
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