WO2018058286A1 - In vitro extraction device and method for peripheral blood mononuclear cells - Google Patents

In vitro extraction device and method for peripheral blood mononuclear cells Download PDF

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Publication number
WO2018058286A1
WO2018058286A1 PCT/CN2016/100293 CN2016100293W WO2018058286A1 WO 2018058286 A1 WO2018058286 A1 WO 2018058286A1 CN 2016100293 W CN2016100293 W CN 2016100293W WO 2018058286 A1 WO2018058286 A1 WO 2018058286A1
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WIPO (PCT)
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assembly
drive
pipetting
axis
connected
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PCT/CN2016/100293
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French (fr)
Chinese (zh)
Inventor
马墨
薛博夫
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深圳市港科深研生物科技有限公司
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Priority to PCT/CN2016/100293 priority Critical patent/WO2018058286A1/en
Publication of WO2018058286A1 publication Critical patent/WO2018058286A1/en

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00Apparatus for enzymology or microbiology

Abstract

Disclosed is an in vitro extraction device (10) for peripheral blood mononuclear cells, comprising a base (100), a circuit control assembly (200), a container assembly (300), a first axis moving assembly (400), a second axis moving assembly (500), a moving frame (590), a pipetting mechanism (600), a drive assembly (700) and a visual sensor assembly (800). The first axis moving assembly (400) and the second axis moving assembly (500) drive the pipetting mechanism (600) to slide via the moving frame (590). The pipetting mechanism (600) sucks up and releases substances under the action of the drive assembly (700). The circuit control assembly (200) calculates the position of the buffy coat according to an image acquired by the visual sensor assembly (800), and controls, according to the position of the buffy coat, the mechanism (600) to suck up the buffy coat.

Description

Peripheral blood mononuclear cells in vitro extraction methods and apparatus FIELD

The present invention relates to the field of medical devices, particularly to a Peripheral Blood Mononuclear Cells in vitro extraction apparatus and method of the PBMC extracts.

Background technique

After blood anticoagulated whole blood is called, is divided into three layers after centrifugation of whole blood, from top to bottom is a plasma layer, a buffy coat, and red blood cell layer. Plasma layer mainly comprising plasma, water, protein, salts and other various ion; buffy coat including platelets, lymphocytes, peripheral blood mononuclear cells (PBMC, Peripheral blood mononuclear cell) and granulocytes, the proportion of these cells in close physical , therefore, are gathered in the buffy coat; Young red blood cell layer can be simply divided into red blood cells and normal red blood cells, since both the red blood cells in the different growth periods of slightly different specific gravity.

Wherein the current peripheral blood mononuclear cells in vitro lymphocyte, individualized cancer immunotherapy, stem cell therapy and related studies have important applications, therefore, it will usually extracted for testing, but the current cell nucleus are extracted by the manual operation, which must require the operator to hand dexterity, and the operation is to be skilled because of the white film layer at the time of suction If after several agitation will cause the red blood cells mixed buffy coat layer, thereby reducing the white extraction rate of the film, on the extraction of peripheral blood mononuclear cells. When dilutions were collected manually and the white film layer laid on Ficoll solution, the diluent is added manually operated intensity control is not good, it is likely to impact on the liquid surface of Ficoll solution, and dilution with white film layer Ficoll the mixed solution, resulting in a change in the density of Ficoll solution, according to theoretical density to make the right separation of peripheral blood mononuclear cells after centrifugation subsequent impact, thus affecting the purity of peripheral blood mononuclear cells.

SUMMARY

Based on this, it is necessary to provide a higher extraction rate in vitro peripheral blood mononuclear cells, higher purity extract of peripheral blood mononuclear cells of peripheral blood mononuclear cells of the extraction device.

In addition, also provides an in vitro method of extraction of peripheral blood mononuclear cells.

One kind of peripheral blood mononuclear cells in vitro extraction device, comprising:

Base;

A control circuit assembly is fixedly mounted on the base;

Container assembly fixedly mounted on said base, said container assembly comprises a plurality of containers, each container of said plurality of means for supporting the sample, diluent and lymph separated liquid;

Moving the first shaft assembly fixed to said base assembly and connected to the electrical control circuit, wherein said control circuit controls operation of the first shaft assembly moving assembly;

Moving the second shaft assembly, fixedly mounted on said first shaft moving assembly, and the assembly is electrically connected to the control circuit, wherein the first drive shaft assembly to move said second shaft moving assembly along a first axis move, and the control circuit controls operation of the second shaft assembly moving assembly;

Moving frame fixed to the moving assembly of the second shaft, said second shaft such that movement of the drive assembly to move the slide frame along the second axis;

Pipetting means, along the third shaft slidably mounted to the moving frame, so that the pipetting means capable of extending into the container, and able to draw substance and a releasable substance, said pipette mechanism and the a control circuit assembly electrically connected to said circuit control means controls said working fluid shift mechanism;

A drive assembly connected to the pipetting means, and electrically connected to said circuit control assembly, the drive assembly for providing a force to said material, and a suction force of a release material pipetting mechanism, wherein said driving circuit control means controls said working assembly;

Vision sensor assembly along a third axis parallel to said fourth shaft slidably mounted to the moving frame, said visual sensor assembly and control assembly electrically connected to said circuit, said visual sensor assembly for collecting the said image of the sample container, and the image can be transmitted to said circuit control module, said control circuit assembly can calculate the position of the white film of the sample according to the image, and can be in accordance with the position control of the white film of the first shaft moving assembly, moving said second shaft assembly, said drive assembly and said pipetting mechanism operates to cause the suction of the sample pipetting mechanism in white film.

One kind of peripheral blood mononuclear cells in vitro extraction method, comprising the steps of:

Providing the above-described in vitro peripheral blood mononuclear cells extracting means;

The diluted fluid, lymph fluid and separating the three samples were loaded into a container;

Said circuit control module controls said first shaft and said second shaft moving assembly moving at least one working assembly, so that the moving frame and the drive mechanism of the pipetting assembly to move the visual sensor, then the circuit control means for controlling the movement of the visual sensor assembly along said fourth axis slide frame, so that the vision sensor at a bearing assembly to move the container containing the sample, said control circuit controls said assembly vision sensor assembly acquiring images of the sample, the visual sensor assembly of the image transmitted to said circuit control module, said control circuit assembly of the sample is calculated according to the white layer of the image using the following formula position, wherein, assuming that P represents a point in space, defining the point P in the second coordinate axis and the coordinate system established as a third shaft (PAx, PAy), defining the point P the two-dimensional coordinate visual sensor coordinate system of the image acquisition assembly is located in the range (PCx, PCy), defining the original coordinate system of the image acquisition vision sensor assembly is located Point, and the coordinate origin of the second coordinate axis and the fourth axis to establish a coordinate system (OCx, OCy) defining the second axis of the fourth shaft to establish a second axis and the coordinate system is established in a third coordinate axis (OBx, OBy), R1, R2, R3 and R4 each represent a space between the rotation axis as transformation, scaling and translation factor:

Figure PCTCN2016100293-appb-000001

Said circuit controls the pipetting mechanism assembly along said third axis slide position control of the white film layer calculated, and the pipetting means extending into the vessel containing the sample up to the calculated the position of said white film layer, said circuit control means further controls the drive assembly to the pipetting means providing a suction force to cause the substance pipetting mechanism draw the white film of the sample;

The control circuit controls the pipetting assembly means slidable along said third axis inverted, so that the pipetting means is removed from the vessel containing the sample, said circuit control means controls said first axis assembly and the second shaft moving in at least one working assembly, so that the moving frame driving the pipetting means slidably fitted to said container at a diluent, said circuit control means controls the pipetting mechanism in the third axis slide until the slide to a container containing the diluent, the circuit control module controls said drive assembly work, the pipetting means to send a force release of the substance albuginea injection layer containing said diluent vessel mixing, dilution to obtain a white film layer;

The control circuit assembly for controlling the movement of said pipetting mechanism frame slides along the third axis, so that the pipetting means into said white film layer diluent, said circuit control means controls the drive assembly to the pipetting means providing a suction force to cause the substance pipetting mechanism draw the diluent white film layer;

Said circuit control module controls said first shaft and said second shaft moving assembly moving at least one working assembly, to drive the movement of the slide frame to move the pipetting means to the bearing containing the isolated lymph the liquid container, and said circuit control means controls the pipetting mechanism of the mobile frame along the third axis, and controlling the drive of the pipetting assembly to provide a means to force a release of a substance the buffy coat was diluted into the container with the bearing lymph separated liquid, and finally by centrifugation to obtain peripheral blood mononuclear cells.

Extracting means above in vitro peripheral blood mononuclear cells to control operation of the various components of the circuit by the control means, the first shaft and the second shaft moving assembly driven carriage assembly slidably moving the pipetting mechanism and the visual sensor and the first shaft assembly sliding second shaft, a third shaft pipetting mechanism to slide along the pipetting mechanism can extend into the vessel, and a driving motor to provide a force mechanism pipetting substance and a releasable substance suction force to pipetting mechanism able to draw substance and a releasable substance, circuit control means controls each component according to the position of the buffy coat collected image calculation obtained vision sensor assembly work to draw a sample pipetting mechanism of a white layer, it is possible to accurately determine albuginea position of the layer of the white film layer to extract, help to improve the extraction efficiency of the white film layer, white layer such that the extraction efficiency up to 80%, thus improving the extraction rate of peripheral blood mononuclear cells; and purified by using the above peripheral blood mononuclear cells in vitro the extracting means extracts only the control circuit components disposed corresponding to the operating parameters according to the actual situation than staffing Stable and effectively prevent the separated liquid diluent and lymph not manually control the intensity level of separated liquid lymph impact caused over the white film layer mixture on the purity of the separation of peripheral blood mononuclear cells, and therefore, With the above peripheral blood mononuclear cells in vitro extraction apparatus can be improved purity of peripheral blood mononuclear cells, and more simple operation.

BRIEF DESCRIPTION

In order to more clearly illustrate the technical solutions in the embodiments or the prior art embodiment of the present invention, the accompanying drawings briefly described embodiments or the prior art needed to be used in describing the embodiments. Apparently, the drawings in the following description only some embodiments, those of ordinary skill in the art is concerned, without creative efforts, reference may also be other embodiments of the present invention obtained according to these drawings.

1 is a configuration in vitro embodiment of the peripheral blood mononuclear cells schematic extracting means;

FIG 2 is a peripheral blood mononuclear cells in vitro as shown in FIG. 1 extracts a first position sensor means, attached to the structure of a first state and a second state of the sensor the sensor slide, tube rack and the guide plate assembled with schematic;

FIG 3 is an exploded view of the peripheral blood mononuclear cells in vitro as shown in FIG. 2 slide sliding extracting means;

Another angle of the device 4 is peripheral blood mononuclear cells in vitro as shown in FIG. 1 a schematic view of a structure extracted;

In vitro test tube rack exploded view in FIG. 5 is shown in Figure 3 peripheral blood mononuclear cells of the extraction apparatus;

FIG 6 is a peripheral blood mononuclear cells in vitro as shown in FIG 1 an exploded view of a first extraction device assembly axis;

7 is peripheral blood mononuclear cells in vitro as shown in FIG 1 an exploded view of a second extraction device assembly axis;

The liquid-absorbent structure diagram of FIG. 8 is a cylindrical peripheral blood mononuclear cells in vitro as shown in FIG. 1 the extraction means;

FIG 9 is a peripheral blood mononuclear cells in vitro as shown in FIG. 1 extraction device carriage, exploded view of the slide assembly and the drive assembly;

FIG 10 is a peripheral blood mononuclear cells in vitro as shown in FIG. 1 the extraction means bracket pipetting, liquid-absorbent plug and cylinder assembly take structural diagram assembled together;

FIG 11 is a peripheral blood mononuclear cells in vitro as shown in FIG extraction device 10 pipetting stent, inserting and removing pipetting assembly and an exploded view of the cartridge;

FIG 12 is an exploded view of the drive assembly in vitro peripheral blood mononuclear cells shown in FIG. 1 the extraction device;

13 is another peripheral blood mononuclear cells in vitro as shown in FIG. 12 extraction device drive assembly exploded view;

FIG exploded visual sensor assembly apparatus 14 is peripheral blood mononuclear cells in vitro as shown in Figure 1 extracted;

In vitro 15 is shown in FIG. 8 peripheral blood mononuclear cells extraction device carriage, another perspective exploded view of the slide assembly and the drive assembly;

FIG 16 is a flowchart of a method of extracting an embodiment of the in vitro peripheral blood mononuclear cells.

detailed description

To facilitate understanding of the present invention, with reference to the following drawings related to the present invention will be more fully described. It gives the preferred embodiments of the invention in the drawings. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments of the object is to be understood that the present disclosure more comprehensible comprehensive.

Incidentally, when an element is referred to as being "fixed", which may or may present centering element directly to the other element. When an element is regarded as being "connected", it can be directly connected to another element or intervening elements may be present simultaneously. As used herein, the term "vertical", "horizontal", "left", "right" and similar expressions are merely for illustrative purposes.

Unless defined otherwise, the same meaning as used herein, all technical and scientific terms used with the present invention belongs to the field of the art technology generally understood. In the description herein, the terms used in the present invention only for the purposes of describing particular embodiments and are not intended to limit the invention. As used herein, the term "and / or" includes any one or more of the associated listed items and all combinations thereof.

As shown, in vitro Peripheral blood mononuclear cells to an embodiment of the apparatus 110 for extracting peripheral blood mononuclear cells from whole blood. The peripheral blood mononuclear cells in vitro extraction apparatus 10 includes a base 100, a control circuit assembly 200, container assembly 300, a first axis moving means 400, a second axis moving means 500, moving frame 590, a pipetting mechanism 600, the drive assembly 700 and a visual sensor assembly 800.

Referring to FIG. 2, the base 100 is a whole peripheral blood mononuclear cells in vitro extraction apparatus 10 of the support member. Specifically, the base 100 includes a base plate 110, the guide plate 120 and the slide plate 130.

Plate 110 is substantially rectangular plate.

Referring to FIG 3, the guide plate 120 is parallel to two 120, two guide plates spaced and fixed relative to the base plate 110. Wherein each two guide plates 120 defines a sliding slot (not labeled), and the extending direction of the guide chute two parallel plates 120. In the particular embodiment illustrated embodiment, the guide plate 120 is a strip-shaped plate, the same as the extending direction of the sliding guide plate 120.

The slide plate 130 along the extending direction of the guide plate 120 is slidably disposed between the two guide plates 120 and 132 attached to rail 122 and the chute cooperating on opposite sides of the slide plate 130. In the particular embodiment illustrated, the length of the guide plate 120 is slightly greater than the length of the slide plate 130.

Further, the position sensor 140 is mounted for sensing the position of the slide plate 130 on the backplane 110. Specifically, the position sensor 140 is provided near one end of the guide plate 120, the slide plate 130 is mounted from the other end of the guide plate 120.

Further, one end of the slide plate 130 defines a handle aperture 134. Wherein the slide plate 130 is mounted on two guide plates 120 from the end remote from the handle hole 134.

Referring to FIG. 4, the control circuit assembly 200 is fixed on the bottom plate 110 of the base 100. Circuit and the data processing apparatus 10 controls a circuit assembly 200 for the entire peripheral blood mononuclear cells in vitro extraction means. Specifically, the control circuit 200 includes a circuit assembly 210 and the display control processor 220, processor 210 controls circuits for in vitro whole peripheral blood mononuclear cells of the extraction circuit and the data processing means 10, the display circuit 220 and the control processor 210 is electrically connected to the display 220 through the input parameters to control the entire apparatus. Wherein the position sensor 140 is electrically connected to the circuit control module 200.

The container assembly 300 mounted on the base 100, container assembly 300 includes a plurality of containers 310, 310 for respectively supporting a plurality of containers containing the sample, and lymph separated liquid diluent. Wherein, lymph Ficoll separation liquid may be a solution or the like, for example. Specifically, a plurality of containers 310 are fixed to the slide plate 130. Wherein, the container 310 may be a tube, a reagent bottle and the like. In the particular embodiment illustrated, there are twelve tubes 130 on the slide and two reagent bottles; two reagent bottles are used for installing the lymph and the separation liquid diluent; diluent may be physiological saline or PBS solution; tube They are used for installing the sample, lymph experimental desired separation liquid and the diluent.

Further, the slide 130 defines a groove 136 accommodating the reagent bottle is partially received in the receiving groove 136, the receiving groove 136 is attached to a first state of the sensor 150 for sensing whether the reagent bottle is present. Wherein a first state of the sensor circuit 150 is connected to the control circuit 200. The control circuit assembly processor 210. Specifically, two receiving slot 136, a reagent bottle corresponding to a respective receiving groove 136.

Referring to FIG 5, a further container assembly 300 further includes a tube holder 320 for placing the tubes, tube rack 320 is detachably mounted to the slide plate 130. Wherein a test tube rack 320 includes a positioning plate 322, side panels 324 and 326, the positioning plate 322 and the slide plate 130 secured detachably connected to a plurality of first positioning holes 322a defines the positioning plate 322; the panel 324 defines a plurality of the second positioning holes 324a, 324a and a second positioning hole in a first position and a position of the positioning holes 322a correspond; one of the two side plates 326, 326 respectively at both ends of a side plate 322 of the positioning plate, panel One end 324 fixedly connected to both ends of the other side plate 326, respectively, and the other end of the positioning plate 322, the other end of the panel 324 is fixedly connected. A second tube passes through a first positioning hole 324a and positioning hole 322a is positioned in the tube rack 320. Wherein a test tube rack 320 close to the receiving groove 136 is provided.

Please Referring again to FIG. 3, further, the slide plate 130 is also mounted on a second plurality of state sensor senses the presence or absence of the cuvette 160, the processing control circuit of each inductor 160 and the second state circuit 200 of the control assembly 210 is electrically connected to a second state of the sensor 160 corresponding to a first positioning hole 322a.

Further, the positioning plate 322 further defines a third positioning hole 322b, the positioning hole 322b is provided with cooperating third positioning post 138, post 138 is positioned through the third positioning hole 322b is provided on the slide plate 130 and the test tube rack 320 130 is positioned on the slide.

Referring to FIG 6, a first axis moving means 400 is fixed on the base 100, and a control circuit connected to the circuit assembly 200 electrically control processor 210. Wherein the control circuit components of the circuit 200 of the control processor 210 controls the first shaft 400 of the moving assembly work. Specifically, a first axis moving means 400 is fixed on the base plate 110. Wherein the first shaft 400 includes a first beam moving assembly 410, a first motor 420, a first roller 430, a first belt 440 and the first slider 450.

A first cross member 410 is fixed on the bottom plate 110 of the base 100. Specifically, a first cross member 410 extending along a first axis; ends of the first cross member 410 by two bottom plate 110 and the base 100 of the bracket 460 is fixedly connected, a first cross member 120 and 410 disposed close to a guide plate. In the particular embodiment illustrated embodiment, the extending direction of the first cross member 410 extending in the direction parallel to the guide plate 120; a first beam and the guide plate 410 and the length 120 is substantially equal.

The first motor 420 is fixed to the first cross member 410, a first motor 200 and the electric circuit components of the control circuit 420 and control processor 210 is connected to a first motor 200 controls the circuit control module 420 work. Specifically, the first motor 420 is located on the end opposite the base plate 410 and the side 110 of the first cross member.

The first roller 430 is two. Wherein a first roller 430 connected to the first drive motor 420, the other first roller 430 is rotatably mounted on the first beam 410. Specifically, the other of the first roller 430 is located remote from the first end of the first cross member 410 of the motor 420.

The first timing belt 440 arranged in two sets of the first roller 430, so that the first timing belt 440 is connected to the two first drive rollers 430. Wherein the first motor 420 driving a first roller 430 is rotated, the timing belt 440 to drive the first movement, and the other of the first drive roller 430 is rotated.

The first slider 450 is fixed on the first timing belt 440, so that the first timing belt 440 drives the first slider 450 along a first axis slide.

Further, the first shaft assembly 400 further includes two moving a first end cap 470 and the first cover 480, the two first end caps 470 are fixed to the ends of the first cross member 410, the two first roller 430 rotatably received in the first two end caps 470, 480 at both ends of the first cover 470 are fixedly connected to the two first end cap, a first shield plate 480, the first belt 440 and the first slider 450.

Referring to FIG 7, the second shaft 500 is fixedly mounted to the moving assembly moves the first shaft assembly 400, and electrically connected to the circuit control module processor 210 controls circuits 200, wherein the first axis of the moving assembly 400 can be driven two-axis moving means 210 controls the operation of the second shaft 500 moving assembly 500 along a first axis, a control circuit 200 of the control processor assembly circuit. Specifically, the second shaft assembly 500 moves the first shaft assembly 400 moves the first slider 450 is fixedly connected. Wherein the moving assembly 500 includes a second axis second beam 510, a second motor 520, second roller 530, a second timing belt 540 and the second slider 550.

Second beam 510 with the first shaft assembly 400 moves the first slider 450 is fixedly connected to the first moving assembly 400 to drive the second cross member 510 along a first axis. Specifically, the second cross member 510 extending along a second axis; a second axis perpendicular to the first axis; 120 perpendicular to the guide plate 510 and the second beam.

Further, the first shaft assembly 400 further includes a first mobile moving block 485 and the two first guide rails 490, 485 of the first moving block 450 is fixedly connected to the first slider, the first two 485 defines a first moving block mating groove (not labeled); the two first guide rails 490 disposed in parallel and opposite sides of the first belt 440, and extends along the sliding direction of the first slider 450, the two first rail portions 490 are received in the two first mating groove, so that the first moving block 485 is slidable on the first guide rail 490. Wherein the first moving block 450 and the first cover 485 is located between the first slider 480; second cross member 510 is fixed to the first moving block 485 is connected to the second cross member 510 with the first shaft 400 connected to the moving assembly more stable.

Further, the first shaft assembly 400 further includes a first moving sliding base 495, 485 of the first sliding base 495 is fixedly connected to the first moving block, a first plate 480 positioned between the first sliding base 495, and a first moving block 485 , the second beam 510 with the first sliding base 495 is fixedly connected.

A second motor 520 and a second cross member 510 is fixedly connected, a second circuit 520 and the motor control circuit assembly 200 control processor 210 is electrically connected to the second motor control circuit 210 control processor 200 control assembly work. Specifically, the second motor 520 is located near one end of the second cross member 510 of the second shaft 500 of the moving assembly.

Second roller 530 is two, wherein the second roller 530 and a second drive motor 520 is connected to the other of the second roller 530 is rotatably mounted on the second beam 510. Specifically, the other second roller 530 located at the second end of the second cross member 510 remote from the motor 520.

The second timing belt 540 is sleeved on the two second rollers 530, 540 so that the second timing belt 530 is connected to the two second drive rollers. Wherein the second motor 520 drives a second roller 530 rotates, the timing belt 540 to drive the second movement, and a second drive roller 530 is rotated further.

The second slider 550 is fixed to the second timing belt 540, so that the second timing belt 540 drives the second slider 550 slides along the second axis.

Further, the second shaft moving assembly 500 further includes two second end cap 560 and a second cover plate 570, two second end cap 560 are fixed to the ends of the second cross member 510, the two second rollers 530 rotatably housed within the two second end cap 560, both ends of the second cover 570 are fixedly connected to the two second end cap 560. Wherein the second motor 520 is fixed to the second cover plate 570.

Referring to FIGS. 1 and 4, the second moving frame 590 is fixed to the moving assembly 500 to move the second assembly 500 can drive the second carriage 590 along the axis slide. Specifically, the carriage 590 and the second slider 550 is fixedly connected.

Please refer again to Figure 7, a further, second shaft moving assembly 500 further includes a second moving block 580 and a second guide rail 585, a second moving block 580 and the second slider 550 is fixedly connected, a second moving block 580 defines two second mating grooves (not labeled), two second guide rails 585 disposed on opposite sides of the second timing belt 540, and extends along a second axis, the two second guide rails 585 are respectively received in the two second portions mating groove, so that the second moving block 580 is slidable on the second guide rail 585. Wherein the second moving block 580 is located between the second slider 550 and a second cover 570. Wherein the second moving frame 590 and the moving block 580 is fixedly connected.

Further, a second carriage 590 and 580 fixed to the moving block 586 connected by two webs, each web 586 and the second end of the moving block 580 is fixed, and the other end of the moving frame 590 is fixedly connected.

A pipetting mechanism 600 along the third shaft slidably mounted on the carriage 590, so that the pipetting mechanism 600 can extend into the container 310, and be able to draw substance and a releasable substance. I.e. pipetting mechanism 600 can be a mass transfer vessel 310 to another container 310. Pipetting mechanism 600 and electrically connected to the circuit control module 200 of the processor control circuit 210, the control circuit of the circuit assembly 200 control processor 210 controls the pipetting mechanism 600 of the work. Wherein the third axis is perpendicular to the second axis. In the particular embodiment illustrated, the third axis is a vertical direction.

Referring to FIG. 8, specifically, the pipetting mechanism 600 comprises a liquid-absorbent tube 610.

Pipetting tube along the third shaft 610 is slidably mounted on the carriage 590. Specifically, the absorbent tube 610 comprises a barrel 612, the needle 614 and the cover 616.

Cylinder housing 612 is open at one end, the other end of the cylindrical body 612 defines a fluid suction openings 612a, the cylindrical body 612 along the third shaft slidably mounted to the carriage 590. Specifically, the cartridge 612 further includes an end fixed to the cylindrical body 612, and the liquid through the tubular suction hole 612a of the cylinder 612 is connected to the connection portion 612b, and a connecting portion 612b with the fluid suction openings 612a arranged coaxially.

Needle 614 is fixedly connected with the cylinder 612, and the fluid suction openings 612a communicating. Specifically, one end of the needle 614 is housed in the connecting portion 612b.

Cover 616 is provided on the lid 612 of the cylinder opening and connected to the seal cylinder 612, the cover member 616 defines with a cylindrical body 612 in communication with vent (not shown), one end of the cover body 616 is provided with the cover member 616 is fixedly connected, and the communicating portion communicating with the vent hole tubular 616a.

Referring to FIG. 1 and FIG. 9, a further, peripheral blood mononuclear cells in vitro extraction apparatus 10 further includes a liquid suction tube 610 for driving the carriage 590 along the sliding shaft 620 of the third slide assembly, the slide assembly 620 fixedly mounted on the carriage 590. Wherein the slide assembly 620 includes a motor 621 pipette, pipette roller 622, the timing belt 623 pipette, pipette screw 624, a guide member 625 and the pipette pipetting slider 626.

Pipetting motor 621 is fixed to the moving frame 590, and a control circuit connected to the circuit assembly 200 electrically control processor 210, the control circuit 210 controls the processor circuit 200 of the control assembly 621 to the pipetting operation of the motor.

Pipetting two rollers 622, one roller 622 is connected to the pipette pipette drive motor 621, the roller 622 is rotated another pipette mounted on the carriage 590.

Pipetting belt 623 provided in two sets of rollers 622 on the pipette and pipette connected to two drive rollers 622.

Pipetting screw 624 and the motor 621 away from the pipette pipetting roller 622 is fixedly connected to the screw 624 pipetting with a pipetting roller 622 rotates. Pipetting screw 624 extending along a third axis.

Pipetting guide member 625 fixed to the carriage 590, the pipette extension direction of the guide member 625 and the spindle 624 parallel pipetting.

Pipetting the slider 626 is slidably sleeved on the spindle 624 and pipetting pipetting guide member 625, the liquid suction tube 612 of the cylinder 610 is fixed to the slider 626 pipetting. Specifically, the pipette 626 and the slider 624 threadedly engaged with the threaded pipette. Wherein the motor 621 to drive its pipetting a pipette roller 622 rotates, the timing belt 623 and the sliding pipette, pipette pipette belt drive roller 623 further rotates 622, the screw 624 with another pipette pipetting connected the roller 622 is rotated, the pipette 626 in the pipette slide screw 624 is slid to the slider 626 driven by pipetting the liquid suction tube 610 slides along the third axis 590 on the moving frame.

Further, the slide assembly 620 further includes a pipetting plate 627, the groove 592 defines a stripe-shaped carriage 590, a timing belt 623 and pipetting pipetting threaded bar 624 are received in the groove 592, the cover 627 pipetting the lid groove 592 provided on the strip, the strip slot 592 and the shielding portion.

Referring to FIG. 10, further, a pipetting mechanism 600 further comprises a bracket 630 and pipette assembly 640 inserted and removed. Pipette holder 630 and assembly 640 for inserting and removing pipetting cylinder 610 is fixed to the slide assembly 620 of the slider 626 on the pipette.

Pipette holder 630 is slidably mounted on the carriage 590. Specifically, the pipette holder 630 and the sliding component 620 of pipette 626 is fixedly connected to the slider to enable the sliding bracket 630 on the moving frame 590 pipetting. In the particular embodiment illustrated, the pipette holder 630 comprises a housing 632, an upper cover 634 and lower cover 636, an opening end of the housing 632, the upper cover 634 and lower cover 636 are provided on the housing cover 632 of two openings on.

Referring to FIG. 11, the inserting and removing pipetting assembly 640 is fixed to the bracket 630. Wherein the plug assembly 640 comprises a connecting member taken 641, the resilient member 642 and the adapter tube connector 643.

Connecting member 641 slidably received in the pipette holder 630. Wherein the connector member 641 comprises a plate-like connecting body 641a, 641a connected to one end of the plate-like hollow cylindrical body 641b and fixedly connected to one end of the body 641c connection boss 641a is fixedly connected to the plate-like. A plate connected to the body 641a defines a through hole (not shown), a hollow post through hole 641b, 641c boss homogeneous communication connection, so that the connection boss 641c communicating with the hollow column 641b. Wherein the boss 641c is connected to one end of the body 616 communicates with the socket portion 616a away from the cover, so that the liquid suction tube 610 and the pipette holder 630 fixedly connected to, and communicates with the connector portion 616a is connected through the boss 641c so not only to achieve the liquid suction tube 610 mounted on the pipette holder 630, but also to achieve a liquid-absorbent tube 610 communicates with the connecting member 641.

An elastic member 642 accommodated in the pipette holder 630, the end of the elastic member 642 and the pipette holder 630 elastically abuts the other end of the connecting member 641 is connected to the plate-like elastic body 641a abuts to the elastic member 642 connected to member 641 and providing an elastic support force of a resilient restoring force, to prevent excessive back connecting member 641, and the connecting member 641 can be reset. Specifically, two of the elastic member 642; 642 and the elastic member is a spring.

Further, the insert and withdrawal assembly 640 further includes a lubricating sleeve 644, sleeve 644 is fixedly accommodated lubrication to the stent pipette 630, open bars connecting member 641b 641 is slidably disposed through sleeve 644 lubrication. I.e. lubricating sleeve 644 to ensure that the connection member 641 smoothly slides within the pipette holder 630.

Adapter tube connector 643 is fixedly mounted on the pipette holder 630, and partially accommodated within the pipette holder 630, the adapter 643 and the tube connector 641 connected to the hollow cylindrical member 614 away from the plate-shaped end of the body 641a of the socket connector, tube connector so that the adapter 643 communicates with the hollow cylinder 614, i.e., to achieve the communication adapter 643 and the tube connector 610 of the liquid suction tube. Specifically, the lubricating sleeve 644 at the transfer pipe-shaped connector 643 and the connector 641 is connected between the body 641a.

Take the insertion assembly 640 to achieve pipetting tube 610 is fixed as follows:

Fixing the absorbent tube 610: sliding component 620 of the slider 626 driven by the pipette moving bracket 630 slides downward, the inserting and removing pipetting assembly 640 with the slide holder 630, until the plug connector component taken connecting stud 640 is inserted into the cap 641c 641 616 communication unit 616a, the elastic compression member 643 to be fully inserted into the connection boss 641c communicating portion 616a, the slider 626 stops sliding pipette, pipette holder 630 so that the stop slide, the slider 626 and then reverse pipetting sliding, to drive the pipette holder 630 slides backward, by an elastic member 630, the connecting member 641 is reset at this time, the liquid suction tube 610 is fixed to the inserting and removing the assembly 640.

Further, in order to achieve automatic separation of the absorbent tube 610, inserting and removing the motor assembly 640 further includes a separation 645, connecting post 646, 647 separated from the slider, the connecting rod 648 and the sleeve 649 separate.

Separating motor 645 fixed to the pipette holder 630, the motor circuit 645 and a control circuit assembly 200 of the separation control processor 210 is electrically connected to the control circuit 210 controls the processor circuit 200 of the control assembly 645 of the separation motor operation.

Connecting post 646 is rotatably housed in the housing 632 of the pipette holder 630, the motor 645 connected to one end of the separation column 646 is connected to the drive.

Separating slider 647 is accommodated in the pipette holder 630, the separation of the slider 647 along the extending direction of the connecting post 646 is slidably sleeved on the connecting post 646. Specifically, the slider 647 and connected to the separation column 646 is screwed.

One end of the connecting rod 648 is accommodated in the pipette holder 630 and the slider 647 is fixedly connected to the separation.

Separating sleeve 649 located outside the pipette holder 630, separating sleeve 649 fixedly connected to one end of the connecting rod 648 away from the slider 647 is separated, so that separation of the connecting rod 648 can drive the slide sleeve 649. Wherein, when the liquid suction tube 610 communicating lid portion 616a of the sleeve 616 is provided on the connecting stud 641 c, separating sleeve 649 is positioned between the bracket 630 and the liquid-absorbent cover cylinder pipetting 616 610, sleeve 649 can abut separation push the lid member 616 so that the liquid suction tube 610 and the connecting member 641 separation.

The method of the insertion assembly 640 to take effect separation of the liquid suction tube 610 is as follows:

Drive motor 645 connected to the separation column 646 is rotated, so that the slider 647 slides on the separate connecting posts 646 and 648 slidably driven by a connecting rod, the connecting rod 648 to drive sleeve 649 towards the separation next to the suction cylinder 610 of the cover 616 of the sliding direction, and the absorbent cartridge 610 pushes against cap 616, the liquid suction tube 610 until the cover 616 is separated from the communication portion 616a and the connecting boss 641 c, thereby achieving separation of the liquid suction tube 610 and the assembly 640 is inserted and removed.

See again FIG. 10, further, pipetting the housing defines a stripe-shaped cutout 632 of the bracket 630 632a, 632a from notch bar near the middle of the housing 632 remote from the end cover 634 extends into the housing 632, the insert and withdrawal assembly 640 further comprises a transmitting means (not labeled) and two position sensors 30 and the position sensor 30 is electrically connected to the circuit control processor 210 is electrically location sensor 30 and the control circuit group 200 is connected to a position sensor 30 fixed to the pipette holder 630 of the cover 636, the other position sensor 30 is fixedly connected to the separation of the slider 647, and the separating position sensor 30 with the slider 647 slides along the strip-shaped notch 632a, and the position of the two inductors 30 abut, when the two abutment position sensor 30, the control circuit means to a feedback circuit assembly 200. the processor 210 controls signal transmission contacts.

Please Referring again to FIG 1 and FIG 3, further, the slide plate 130 is also installed for placing a plurality of liquid suction tube 610 supplies frame 139. Inserting and removing the separated liquid absorbent component 640 can be placed in tube 610 supplies frame 139. Specifically, the consumable holder 139 is removably secured to the slide plate 130. In the particular embodiment illustrated, the slide plate 130 is mounted on a plurality of positioning status button 170, and a third sensor 180, a plurality of positioning bracket 139 and supplies buckle 170 fixed to the slide 130, the third state of the sensor 180 for is there a liquid suction tube 610 on the sensor 139 supplies frame, with the separated liquid suction tube 610 into rack 139 supplies.

Referring to FIGS. 12 and 13, drive assembly 700 is connected to the pipetting mechanism 600, and a control circuit connected to the circuit assembly 200 electrically control processor 210, drive assembly 700 to the pipetting mechanism 600 for providing a suction substance force and the force of a release material, the control circuit of the circuit assembly 200 control processor 210 controls the drive assembly 700 of the work. Specifically, the use of a connecting pipe (not shown) communicating pipe connector adapter 643 and drive assembly 700 and cylinder 610 in communication with the liquid-absorbent drive assembly 700, drive assembly 700 such that the liquid-absorbent suction tube 610 or the gas input the cylinder 610 to the liquid-absorbent material providing a suction force and a release force material. Wherein the drive assembly 700 includes a mount 710, the barrel 720, the driving motor 730, the drive guide rods 740, 750 and drive rod 760 driving the slider.

Mount 710 is fixedly mounted on the bottom plate 110 of the base 100. Specifically, a mounting groove 712 defined in the mounting seat 710.

Syringe 720 is mounted on the mount 710. Specifically, the syringe 720 is received in the mounting groove 712. Wherein the syringe 720 includes a hollow cylinder 722, piston 724 and plunger 726.

Hollow cylinder 722 having an open end and a closed end, hollow cylinder 722 is fixed to the mounting seat 710, the communication hole 722a is opened on the closed end of the hollow cylinder 722. Wherein the connecting pipe remote from the liquid suction tube 610 and the end cover 616 of the communication hole 722a communicating. Specifically, the connecting pipe remote from the head end of the adapter pipe 643 connected to the communication hole 722a communicating.

The piston 724 is slidably received in the open end of the hollow cylinder 722, the piston 724 and the inner wall of the hollow cylinder 722 sealed connection.

Pusher part 726 is accommodated in the hollow cylinder 722, one end of the push rod 726 is fixedly connected with the piston 724.

Driving motor 730 is fixed to the mounting base 710, and electrically connected to the circuit control module 200 controls the processor circuit 210, the control circuit components of the circuit 210 of the control processor 200 controls the driving operation of the motor 730.

Driving the guide rod 740 fixed to the mounting seat 710, 740 in the axial direction of the hollow cylinder 722 extending in a driving guide bar parallel. Specifically, the drive rod 740 is received in the guide groove 712 is mounted.

One end of the drive rod 730 drives the driving motor 750 is connected to the extending direction of the drive spindle 750 and the hollow cylinder 722 is parallel to the axial direction. Specifically, the drive rod 750 is received in the mounting groove 712.

Driving the slider 760 is slidably sleeved on the drive spindle 750 and a drive rod guide 740, the drive slider 760 is fixedly connected to the push rod 726 away from the end of the piston 724, the plunger driver 726 can drive slider 760 slides, the movement of the piston 724 within the hollow cylinder 722. Specifically, the drive slide mounting groove 760 receiving 712.

Further, the drive assembly 700 further includes a drive plate 770, drive plate 770 disposed on the cover mounting groove 712, and the shield mounting groove 712.

Further, the drive assembly 700 further includes a drive limiting member 780, driving the stopper member 780 is fixedly mounted on the mounting seat 710, and 760 may abut and drive the slider, the slider 760 while preventing the drive continues to slide.

Further, the drive assembly 700 further includes a U-shaped support frame 110 is mounted on the bottom plate 790 of the base 100, two U-shaped support frame 110 is fixedly connected to the arm and the base plate, the mount 790 is fixed to the U-shaped support frame 710 is connected to 790 the two arm portions are connected.

Vision sensor assembly 800 along a fourth axis parallel to the third shaft slidably mounted on the carriage 590, the control circuit electrically processor 210 and the visual sensor assembly 800 is connected to the circuit control module 200, a sensor assembly 800 for visual collection container the image of the sample 310, and the image can be transmitted to the circuit control module 200, wherein the processor circuit 200 controls the control circuit assembly 210 of the image can be calculated according to the position of the sample in the white film layer, and the buffy coat can be in accordance with a first axis position control assembly 400, a second axis moving means 500, the drive assembly 700 and work pipetting mechanism 600 to cause the sample pipetting mechanism 600 is drawn in the buffy coat.

Specifically, the circuit controls the circuit control module 200 of the processor 210 is able to position (1) of buffy coat sample was calculated based on the image using the following formula:

Figure PCTCN2016100293-appb-000002

Wherein, assuming that P represents a point in space, a coordinate system defined point P in the second and third axes is established coordinates (PAx, PAy), the point P is defined in the image acquisition vision sensor assembly 800 where the two coordinate-dimensional coordinate system is (PCx, PCy), the coordinate system of the sensor assembly defining a visual image 800 acquired in a coordinate system where the second shaft and the fourth shaft established (OCx, OCy), defining a second axis the origin of the coordinate system and the fourth axis coordinate system established in the second and third axes is established in (OBx, OBy), R1, R2, R3 and R4 each represent a spatial transformation between the coordinate axes rotate, zoom and pan coefficients.

DETAILED estimation process described above using the following formula:

Since the third axis and the fourth axis parallel, wherein the coordinates of the point P in the coordinate system defined in the second shaft and the fourth shaft in the establishment (PBx, PBy), then the second shaft and the third shaft established coordinate system and the second axis and the fourth axis coordinate system established translation relations exist only, the transformation relationship in the following formula (2):

Figure PCTCN2016100293-appb-000003

Exist translation, rotation and scaling of the relationship between two-dimensional coordinate system of the second axis and the fourth axis coordinate system established vision sensor assembly 800 where the captured image, the transformation relation following equation (3):

Figure PCTCN2016100293-appb-000004

Equation (2) and Formula (3) can be integrated to give the above formula (1).

Wherein determining the above formula (1) R1, R2, R3 and R4 and (OBx, OBy) as follows:

1, to select a point P on the device, the coordinate system of the point P in the second and third axes established is represented as (PAx, PAy), i.e. (PAx, PAy) is a known quantity.

2, the second axis drive assembly 500 by moving carriage 590 to drive the vision sensor assembly 800 along a second axis such that the vision sensor assembly 800 is positioned above the camera object, then along the first visual sensor assembly 800 on the carriage 600 in four-axis movement direction of the slide close to the photo material, wherein the visual sensor assembly 800 to the point P, respectively, for three pictures in three different positions of points (OC1, OC2 and OC3), and at the three points biaxial and the coordinate system established by the fourth shaft are, respectively OC1 (OCx1, OCy1), OC2 (OCx2, OCy2) and OC3 (OCx3, OCy3), due to the visual sensor assembly 800 moves in the displacement of the second shaft the fourth axis and the displacement is determined in the visual sensor assembly 800 moves the process, so OC1 (OCx1, OCy1), OC2 (OCx2, OCy2) and OC3 (OCx3, OCy3) is a known quantity. The position of the captured image to the point P, the point P to find the three images respectively corresponding to a coordinate position in the image, respectively (PCx1, PCy1), (PCx2, PCy2) and (PCx3, PCy3). I.e. OC1 (OCx1, OCy1), OC2 (OCx2, OCy2), OC3 (OCx3, OCy3), (PCx1, PCy1), (PCx2, PCy2) and (PCx3, PCy3) are known amount. Establishing the above formula (1) according to the following equations:

Figure PCTCN2016100293-appb-000005

Figure PCTCN2016100293-appb-000006

Figure PCTCN2016100293-appb-000007

The two above-mentioned two subtraction equations to obtain the following two formulas:

Figure PCTCN2016100293-appb-000008

Figure PCTCN2016100293-appb-000009

Whereby solve for R1, R2, R3 and R4:

Figure PCTCN2016100293-appb-000010

3, finally R1, R2, R3, R4, and OC1 (OCx1, OCy1) into the above equation (1), to give the following calculation formula:

Figure PCTCN2016100293-appb-000011

To calculate (OBx, OBy). Thereby determining the formula (1), R1, R2, R3 and R4 and (OBx, OBy).

Then, in the present embodiment, the position coordinates of the white film layer is the point P, wherein the position of the point P in the image pickup white film layer on the image coordinates (PCx, PCy) is a known quantity, vision sensor assembly 800 and the coordinates of the second shaft and the fourth shaft to establish a coordinate system (OCx, OCy), that is, when moving the visual sensor assembly 800 to move relative to the second shaft and the displacement of the axis relative to the fourth position, i.e., ( OCx, OCy) is a known quantity, and the formula R1, R2, R3 and R4 and (OBx, OBy) has been calculated earlier, also a known quantity, so that (1), the container 310 can be obtained according to the above formula the actual position of the white film layer.

Referring to FIG. 14. Specifically, the visual sensor assembly 800 includes a fixed plate 810 fixed to the housing 820, the visual sensor 830, camera 840 and the mirror 850.

The fixing plate 810 along the fourth shaft slidably mounted to the carriage 590.

Referring to FIG. 8 and FIG. 15, further, in order to achieve the fixed plate 810 is slidably mounted on the carriage 590, extracorporeal extraction of peripheral blood mononuclear cells means 10 further includes a fixedly mounted to the carriage 590, for vision sensor drive assembly along the stationary plate 810 of the fourth shaft 800 slidably drive assembly 900. Wherein the drive assembly 900 includes a drive motor 910, drive roller 920, drive belt 930, drive screw 940, drive 950 and drive guides the slider 960.

Drive motor 910 is fixed to the moving frame 590, and a control circuit connected to the circuit assembly 200 electrically control processor 210, the control circuit 200 of the processor circuit 210 controls the control assembly drive motor 910 work.

Two drive rollers 920, a drive roller 920 which is connected to the drive motor drive 910, the other drive roller 920 is rotatably fixed to the moving frame 590.

Drive belt 930 is sleeved on the two drive rollers 920, and is connected to two drive rollers 920 drive.

Drive screw 940 and drive roller 910 away from the drive motor 920 is fixedly connected, so that the drive roller 920 drives the drive screw 940 is rotated. The drive screw shaft 940 extending along the fourth.

Transmission guide member 950 fixedly mounted on the carriage 590, extending in a direction parallel to the extending direction of the guide member 950 and the drive gear 940 of the screw. Specifically, the guide member 950 is a bar-shaped drive plate.

Drive slider 960 is slidably sleeved on the spindle drive gear 940 and the guide member 950, wherein the vision sensor assembly 810 of the fixing plate 800 is fixed to the slider 960 to the transmission. Specifically, the drive gear 960 of the slider 940 threadedly engaged with the screw. Wherein a drive motor 910 drives its connected drive roller 920 rotates to drive the drive belt 930 moves, the drive belt 930 driven by a further drive roller 920 rotates, the drive screw 940 with the other drive roller 920 is rotated, the drive the drive screw 960 in the slider 940 slides along the fourth axis, so that the slider 960 driven by the drive assembly vision sensor fixing plate 810 of the fourth shaft 800 slides along the moving frame 590, and achieve a visual sensor assembly 800 along a fourth axis slide.

Further, drive assembly 900 further includes a drive plate 970, drive plate 970 disposed on the strip tank cover 592, tank 592 and the shielding portion of the strip. Specifically, the cover plate 970 and the drive gear 770 together with the cover plate 592 shielding stripe groove.

Fixing housing 820 is fixedly connected with the fixed plate 810, and together with the fixed plate 810 is formed with a receiving space having a through-gap light.

Vision sensor 830 accommodated in the accommodating space, and fixedly connected with the fixed plate 810, and a control circuit electrically processor 210 and the visual sensor 830 is connected to the circuit control module 200, a control circuit 210 controls the control circuit assembly 200 of the visual sensor 830 processor work.

Camera 840 is accommodated in the accommodating space, and fixed to the visual sensor 830, camera 840 and the visual sensor 830, a circuit control component circuit for controlling the processor 200 to 210 are electrically connected to the camera 840 for the sample container 310 acquired image , and the image can be transmitted to the visual sensor 830, so that the visual sensor 830 can be transmitted to the image control circuit assembly 200.

Referring again to Figure 3, further, a light source 190 mounted circuit also has a control processor and a control circuit assembly 200 electrically connected to the bottom plate 210 of the chassis 100 110, source 190 provides a suitable brightness when the camera 840 to capture an image . Specifically, the test tube rack 320 close to the light source 190 is provided.

Mirror 850 is fixedly mounted on the fixing plate 810, the mirror position and the position of the light through the notch 850 corresponding to, and positioned below the camera 840 to the mirror 850 reflects the image of the sample container 310 to the camera 840 inside, the camera image of the sample 840 in the container 310 can be acquired. Specifically, the angle of the camera lens 840 and the mirror 850 in the axial direction is 135 °.

Further, the visual sensor 800 further includes a mounting bracket assembly 870, mounting bracket 870 is received in the accommodating space, and fixedly connected with the fixed plate 810, a mirror 850 mounted on the mount 870.

Peripheral blood mononuclear cells in vitro the above-described extraction apparatus 10 has at least the following advantages:

Peripheral blood mononuclear cells in vitro the above-described extraction apparatus 10 controls various components via the circuit control module 200 operates, the first shaft 400 and second shaft moving assembly moving assembly 500 drives the carriage 590 slidably pipetting mechanism 600 and the vision sensor assembly the first shaft 800 and the second slide shaft, a third shaft slidably along pipetting mechanism 600 so that the pipetting mechanism 600 can extend into the container 310, the drive assembly 700 to give a pipetting mechanism 600 provides a suction substance force and a force of release material so that the pipetting mechanism 600 able to draw substance and a releasable substance, a sensor assembly 800 for the visual image of the sample collection container 310, and the image can be transmitted to the circuit control module 200, a control circuit assembly the image 200 can be calculated using the position of the sample in the above formula white film layer, the film can be controlled according to the position of the white pipetting mechanism 600 to draw a sample of white film, thus precisely extracted buffy coat, it will help improve the extraction efficiency of the white layer, the white layer such that the extraction efficiency up to 80%, thus improving the extraction rate of peripheral blood mononuclear cells; and by using the outer Peripheral blood mononuclear cells in vitro extraction means 10 extracts only the control circuit components 200 disposed on the respective operating parameters according to the actual situation, stable than manpower to effectively avoid the poor control of the intensity of manually separating liquid lymph caused by white film layer resulting from the level of impact and lymphatic separated liquid diluent mixture on the purity of the separation of peripheral blood mononuclear cells, and therefore, the above-described in vitro peripheral blood mononuclear cells of the extraction device 10 can be improved peripheral blood mononuclear cells purity, and more simple operation.

As shown, the peripheral blood mononuclear cells in vitro extraction methods to an embodiment, the extraction method using the above 16 peripheral blood mononuclear cells in vitro extraction means extracts, steps of the method are as follows:

Step S21: providing peripheral blood mononuclear cells in vitro the extracting means.

Step S22: The dilution liquid separation and lymph samples were loaded into three containers.

Specifically, the dilution was charged in a vessel or lymphatic vessel separation step was charged specifically: first with reagent bottles are separated liquid diluent or lymph placed in the slider accommodating slot, the control circuit assembly first axis moving means control a second operation of the first motor and the second motor shaft moving assembly so that the mechanism is moved to the pipetting position of the reagent bottle or reagent bottle dilutions lymph separated liquid, circuit control element controls the slide assembly pipetting motor operation, so that the liquid suction tube slides to lymph separated liquid diluent or near the direction of the container, then the circuit control module controls the driving operation of the drive motor assembly to the syringe piston movement, in order to give the absorbent a rear barrel force substance pipette diluent or lymph desired amount of separated liquid, the control means controls the driving circuit stops working assembly, the slide assembly and the control direction to cause the pipetting motor operating fluid from a container towards the suction tube sliding, the control circuit controls the first shaft assembly again moving the second assembly operation of the first motor and the second motor shaft movement assembly to move pipetting mechanism Moving to the top of an empty tube, the circuit control element controls the slide motor assembly drive pipetting work, so that the liquid suction tube slides to lymph separated liquid diluent or near the direction of the container, and then controls the driving circuit control element assembly motor operation, so that the syringe piston, to provide a suction tube to force liquid substance release, the release of the separation liquid diluent or lymph.

Wherein the step of loading the sample vessel is manually operated, the desired amount of sample loading.

Wherein the position of the reagent bottle and the reagent bottle lymph separating liquid diluent respectively provided with two sensors a first state, a first state of the two position sensors diluted reagent bottle and the reagent bottle lymph liquid separated liquid signal to the control circuit assembly, the circuit assembly according to a second motor controls operation of the first motor and a second shaft position signal to control movement of the first assembly axis moving assembly to move the pipetting mechanism to the appropriate location.

Wherein the pipetting work machine slide assembly (e.g., pipetting motor speed, direction of rotation, operating time, etc.) may be used to set the parameters to the circuit control means in accordance with the liquid level of the container; a drive motor assembly working parameters may be set according to the actual amount of the respective reagents required (e.g., the drive motor speed, direction of rotation, operating time, etc.) and to provide a liquid suction tube and the suction force of substance release material forces.

Step S23: the circuit control module controls a moving assembly work of the first shaft and the second shaft moving assembly, to move the frame and the drive mechanism pipetting assembly to move the visual sensor, then the circuit control means controls the movement along the visual sensor assembly frame a fourth axis slide such that the vision sensor assembly to move the container, the assembly supporting the control circuit containing the sample, sample collection control visual sensor assembly image, the visual images transmitted to the sensor module control component circuit, a control circuit assembly in accordance with the image as follows position of formula obtained white film layer of the sample:

Figure PCTCN2016100293-appb-000012

Wherein, assuming that P represents a point in space, a coordinate system defined point P in the second and third axes is established coordinates (PAx, PAy), a two-dimensional definition of the point P in the image acquisition vision sensor assembly is located in the coordinate system of (PCx, PCy), the origin of the coordinate system of the coordinate system is defined where the image acquisition vision sensor assembly and the second shaft built fourth shaft (OCx, OCy), defining a second axis, and the origin of the coordinate system of the fourth axis coordinate system established in the second and third axes is established in (OBx, OBy), R1, R2, R3 and R4 each represent a space between the rotation axes of transformation , zooming and panning coefficient.

Specifically, the circuit control means controls at least one operation of the first axis and a second axis component of the moving assembly, to move the frame and the drive mechanism pipetting assembly to move the visual sensor, then the circuit control means controls the movement of the visual sensor assembly frame slides along the fourth axis, such that the vision sensor assembly to move the container, the assembly supporting the control circuit containing the sample, sample collection control visual sensor assembly image, the visual sensor assembly step to the image transfer control circuit assembly specifically comprises: a control circuit controlling a first axis component of the second electric component of the first motor and a second axis moving means at least one working, to drive the sliding carriage, the pipetting mechanism and the visual sensor assembly with the sliding carriage, such that the vision sensor assembly bearing located above the container containing the sample, then the circuit control module controls the drive motor operating the drive assembly, so that the sliding vision sensor assembly to the container containing the sample bearing outer fourth shaft, then the circuit control module controls the visual sensor assembly camera to take pictures of the sample, by the visual image The sensor assembly is transmitted to the control circuit.

Specifically, the circuit control module controls the first axis or the second axis component assembly work, or the first control shaft and the second shaft moving assembly moving assembly work is mainly determined according to the location of the mobile frame.

Wherein the first motor assembly a first axis, a second drive motor drive assembly and a second motor shaft movement assembly may be determined according to the position of the sample container containing the sample bearing parameters (e.g., rate of rotation of the drive motor direction of rotation and rotation time, etc.) of the motor drive control operation.

Step S24: circuit control means controls the position of the white film layer calculated pipetting mechanism slides along the third axis, and the shift mechanism extends into the container containing the sample solution until a white film layer to the calculated position, then the circuit control module pipetting assembly to control the driving means providing a suction force to cause material to draw a sample pipetting mechanism of a white film layer.

Specifically, in step S24: position of the white film layer circuit control means in accordance with step S23 calculated shift control mechanism of the slide assembly was pipetted motor operation, so that the suction tube extending into the container containing the sample solution until the calculation the position of the white film layer, then the circuit control module further controls the drive motor to the drive assembly to provide a cylindrical absorbent material suction force so that the liquid suction tube to draw the sample buffy coat.

Specifically, the pipette pipetting tube means extend into the vessel containing the sample is drawn in the white film sample method is specifically: the pipetting mechanism pipetting cylinder to draw concentric circles moving white film layer; or, pipetting mechanism pipetting tube along an Archimedes spiral motion to draw a white film layer; alternatively, the pipetting tube was made Z-shaped movement mechanism to draw the white film layer.

Step S25: circuit control means controls reverse pipetting means to slide along the third axis, such that the pipetting means is removed from the vessel containing the sample, a control circuit controls the first assembly and the second shaft axis moving means moving at least one working assembly at the container, so that the carriage drive mechanism slides to pipetted containing diluent, the circuit control means controls the pipetting mechanism slides along the third axis, until the slide to the vessel containing the diluent, the control circuit controls the driving assembly work assembly , pipetting mechanism to send a force release of the substance into the container in the buffy coat mixed with diluent to obtain a white film layer dilutions.

Step S26: The control circuit assembly pipetting mechanism moves along a third axis slide frame, so that diluent pipetting means extending into the white film, the circuit control means controls the pipetting mechanism drive assembly to provide a suction substance the force to draw diluent pipetting means white film layer.

Step S27: control means controls the first circuit component and the axial movement of the second axis at least one working assembly, to drive the carriage to slide pipetting means to move the container supporting the separated liquid containing lymph, then the circuit control module pipetting means movement control frame along a third axis, and control means controls the driving circuit assembly provides a force to the release of substances pipetting mechanism to the white film layer containing the diluent into the container bearing lymph separated liquid, Finally by centrifugation to obtain peripheral blood mononuclear cells.

Specifically, in step S27, according to the position of the carriage circuit for determining the first control component is a control shaft moving assembly work, assembly work or the second axis, the first axis or the control assembly and the second shaft moving assembly work long as it can drive the carriage to slide pipetting means to move the container to the supporting separated liquid containing lymph.

Specifically, the circuit control means controls the pipetting mechanism moving frame along a third axis, and control means controls the driving circuit assembly provides a force to the release of substances pipetting mechanism to dilution with white film layer bearing lymphatic injection step separated liquid container in particular: a circuit control means controls the pipetting mechanism along a third axis frame moves the pipetting mechanism until the end of the liquid level from the isolated lymph liquid at a position 0.5 ~ 1mm high, the circuit control means operates to control the speed of the drive assembly of the buffy coat dilution was 0.01ml / s ~ 0.3m / s lymph vessel is filled with supporting liquid for separating, so that the diluent white floating layer was separated lymphoid on the surface, until the buffy coat of diluent injection 3 ~ 5ml, at a rate of 0.3ml / s ~ 0.8m / s injecting dilutions remaining white film layer, and finally by centrifugation to obtain peripheral blood mononuclear cells .

More specifically, the circuit control module controls at least one operation of the first axis and a second axis component of the moving assembly, to drive the sliding carriage to drive the carriage to move the slide mechanism to move liquid separation liquid bearing container containing lymph at, then the circuit control means controls the pipetting mechanism along a third axis frame moves the pipetting mechanism until the end of the liquid from the high level of separated liquid lymph 0.5 ~ 1mm at the position, the control means controls the driving circuit assembly work specific steps of: at least a first operation circuit control means controls the second motor assembly axis moving the first and second axis moving motor assembly to drive the movable frame slid pipetting mechanism, so that the liquid suction tube to move with the upper bearing lymph separated liquid container, then the circuit control module controls the motor of the slide assembly pipetting work, so that the third pipette axis slide mechanism along the cylinder liquid-absorbent, when the end of the liquid from the pipetting mechanism separating lymph position 0.5 ~ 1mm high liquid level, the circuit control module controls the drive motor operates the drive assembly, the drive assembly so that the liquid suction tube to provide a release material Force.

Wherein the first motor and the second motor operating state can be set according to the corresponding parameters required for the operation of the container and the actual position of the pipetting mechanism; pipetting operation state of the motor according to the liquid level of the absorbing liquid is separated from lymph the actual position of the cartridge setting parameter, e.g., pipetting motor speed, rotation time and the like; a drive motor operating state can absorb or release an amount of substance to set parameters based on the desired speed and time.

Wherein the separation step is specifically: the injection of the separated liquid diluent lymph white film layer separated by a centrifuge, such that four hierarchical separation liquid, and top to bottom are a layer of plasma, mononuclear cell layer, lymph The layers were separated and the red blood cell layer, and then drawn mononuclear cell layer.

In vitro extraction methods described above peripheral blood mononuclear cells of peripheral blood mononuclear cells extracted by the extraction means peripheral blood mononuclear cells in vitro, the operation is very simple, high extraction rate of peripheral blood mononuclear cells, and higher purity.

The technical features of the above embodiments of the embodiments can be arbitrarily combined for brevity of description, not for technical features of each of the above embodiments are all possible combinations will be described, however, as long as the combination of these features is no contradiction, It should be considered to be the scope disclosed in the present specification.

The above embodiments are only expressed several embodiments of the present invention, and detailed description thereof is more specific, but can not therefore be understood to limit the scope of the invention. It should be noted that those of ordinary skill in the art, without departing from the spirit of the present invention, can make various changes and modifications, which fall within the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.

Claims (20)

  1. One kind of peripheral blood mononuclear cells in vitro extraction device, comprising:
    Base;
    A control circuit assembly is fixedly mounted on the base;
    Container assembly fixedly mounted on said base, said container assembly comprises a plurality of containers, each container of said plurality of means for supporting the sample, diluent and lymph separated liquid;
    Moving the first shaft assembly fixed to said base assembly and connected to the electrical control circuit, wherein said control circuit controls operation of the first shaft assembly moving assembly;
    Moving the second shaft assembly, fixedly mounted on said first shaft moving assembly, and the assembly is electrically connected to the control circuit, wherein the first drive shaft assembly to move said second shaft moving assembly along a first axis move, and the control circuit controls operation of the second shaft assembly moving assembly;
    Moving frame fixed to the moving assembly of the second shaft, said second shaft such that movement of the drive assembly to move the slide frame along the second axis;
    Pipetting means, along the third shaft slidably mounted to the moving frame, so that the pipetting means capable of extending into the container, and able to draw substance and a releasable substance, said pipette mechanism and the a control circuit assembly electrically connected to said circuit control means controls said working fluid shift mechanism;
    A drive assembly connected to the pipetting means, and electrically connected to said circuit control assembly, the drive assembly for providing a force to said material, and a suction force of a release material pipetting mechanism, wherein said driving circuit control means controls said working assembly;
    Vision sensor assembly along a third axis parallel to said fourth shaft slidably mounted to the moving frame, said visual sensor assembly and control assembly electrically connected to said circuit, said visual sensor assembly for collecting the said image of the sample container, and the image can be transmitted to said circuit control module, said control circuit assembly can calculate the position of the white film of the sample according to the image, and can be in accordance with the position control of the white film of the first shaft moving assembly, moving said second shaft assembly, said drive assembly and said pipetting mechanism operates to cause the suction of the sample pipetting mechanism in white film.
  2. The apparatus according to claim 1, wherein said first axis moving assembly comprising:
    A first cross member fixed to said base;
    A first motor fixed to the first cross member, and electrically connected to the control circuit assembly, wherein said control circuit assembly for controlling the operation of the first motor;
    Two first roller, said first roller wherein a first motor connected to the drive, the other of the first roller rotatably mounted on said first cross member;
    A first timing belt, two sleeved on said first roller, said first timing belt connected to both of said first roller drive;
    A first slider, fixed to the first belt, so that the first timing belt drives the first slider slidable along said first axis, the second axis and the first movable component fixedly connected to the slider.
  3. The apparatus according to claim 1, wherein said second shaft moving assembly comprising:
    Second beam, fixedly connected to the first shaft moving assembly, wherein said first shaft drives said second beam moving assembly along said first axis;
    A second motor fixed to the second cross member, and electrically connected to the control circuit assembly, wherein said control circuit assembly for controlling the operation of the second motor;
    Two second roller, said second roller and wherein a second motor drivingly connected to the other of said second roller is rotatably mounted on the second beam;
    A second timing belt, two sleeved on said second roller and said second roller is connected to two transmission;
    A second slide fixed to the second belt, such that the second timing belt driven by said second slider slidable along said second axis, wherein said moving frame and the second carriage block fixedly connected.
  4. The apparatus according to claim 1, characterized in that said pipetting tube means comprises a liquid-absorbent, the liquid suction tube along the third shaft slidably mounted to the mobile frame, the liquid suction tube the other end comprises a barrel, a needle and a cover, the housing is open at one end of the cylindrical body, the cylindrical body defines a fluid suction hole, along said third axis of said cylindrical body slidably mounted on the moving rack, the needle is fixedly connected with said cylinder, and said fluid suction openings communicating the lid member is provided to cover the opening of the cylindrical body, and sealingly connected to the cylinder, the said cover body defines a vent hole communicating with the cylindrical body is connected; wherein said vent hole and a connecting pipe communicating said drive assembly, and the liquid-absorbent assembly in communication with the drive cylinder, so that the said drive assembly of said cartridge and to the suction or delivery of a gas cartridge to provide a liquid-absorbent suction force substance or a substance releasing force.
  5. The apparatus according to claim 4, characterized by further comprising means for driving the movement of the liquid absorbing cartridge frame along a third axis of said slide assembly to slide, said slide assembly comprising:
    Pipetting motor, fixed to the moving frame, and electrically connected to said circuit control assembly, wherein said circuit control means controls the pipetting operation of the motor;
    Pipetting two rollers, wherein one of said roller and said pipette pipetting motor drivingly connected, the other transfer roller rotatably mounted to said fluid displaceable frame;
    Pipetting belt, provided two sets of the rollers on the pipette and the pipette is connected to two wheel drive;
    Pipetting screw, and the motor away from the pipette pipette roller fixedly connected to the pipette with a pipette roller screw is rotated, and the pipette along the third axis lead screw extend;
    Pipetting guide member fixed to the moving frame, extending in the shift direction of the liquid guide member and the shift spindle extending parallel to the direction of liquid;
    Pipetting slider slidably disposed in said sleeve and said screw pipetting pipetting upper guide member, wherein the pipetting tube is fixed to the barrel shifter on the liquid slider.
  6. The apparatus according to claim 4, wherein said mechanism further comprises a pipette assembly and fixing bracket inserting and removing pipetting assembly, the pipette holder slidably secured to said moving frame, said insertion taking on the pipetting bracket, and for the cylinder is fixed to the absorbent liquid on the shift bracket, inserting and removing the assembly comprising:
    Connecting member is slidably received in said shift the liquid holder, the connecting member comprises a plate-like connecting body connected at one end fixedly connected to the hollow stud body and the plate-like end connected to the plate-like body fixedly connected hollow connecting studs, the through hole and the hollow column, the heterogeneous communication connection boss, so that the connection boss in communication with said hollow cylinder;
    An elastic member, the fluid received in the shift bracket, the end of the elastic member with the elastic contact pipette holder, the other end of the plate-like body is connected to the connecting member elastically abutting on the elastic member said connecting member to provide an elastic supporting force and an elastic restoring force;
    Transfer pipe connector, fixedly mounted to move on the liquid holder, and partially received in the shift of the liquid holder, the adapter tube connector away from the plate-shaped hollow column connected to the connector body One end of the socket, so that the transfer pipe is connected to the hollow cylinder head in communication, the adapter is connected to the tube connector tube end remote from the drive assembly in communication;
    Wherein the absorbent is provided on the cartridge lid member fixedly connected to one end of the cover, and a tubular communicating portion communicating with the venthole, the other end of the communicating portion to the socket for connecting projection column, and the communicating convex portion sleeved on said column connected to said liquid suction tube is fixedly connected to said connector member, and said communicating portion and the communicating connection studs.
  7. The apparatus according to claim 6, wherein said plug assembly further includes a lubricating sleeve taking the lubricating sleeve is fixedly received in the inner carrier pipette, hollow post of the connector is slidably disposed through in the lubricating sleeve.
  8. The apparatus according to claim 6, wherein said plug assembly further comprises taking:
    Separation motor fixed to the shift on the liquid holder, and the separation motor control assembly electrically connected to said circuit, said circuit control means for controlling the operation of the motor of the separation;
    Connecting post, is rotatably received in the inner pipette holder, one end of the connecting columns motor drive connected with the separator;
    Separating slider housed in said inner pipette holder connected to the separation column direction extending along the slide sleeve slidably disposed in said connecting post;
    A connecting rod having one end received in the pipette within the housing and fixedly connected to the separating slide;
    Separating sleeve, located outside the liquid shift bracket, one end of the separation sleeve remote from the connecting rod is fixedly connected to the separating slider, so that the separating sleeve can be slid over the connecting rod;
    Wherein, when the communication unit is connected to the sleeved convex column, separating the sleeve and is located between the pipette holder body said lid, capable of resisting the separating sleeve so that the cover said liquid-absorbent member connected to said cylinder and separated.
  9. The apparatus according to claim 4, wherein said drive assembly comprising:
    Mount, fixedly mounted on the base;
    Syringe, comprising a hollow cylinder, the piston and the push rod, said hollow cylinder having an open end and a closed end, said hollow cylinder is mounted on the mounting seat, defines a closed end of the communication hole of the hollow cylinder, the a piston slidably received in the open end of the hollow cylinder, the piston and the inner wall of the sealing hollow cylinder is connected, the push rod portion received in the hollow tube, one end of said push rod and said piston fixed connection, wherein the connection tube end remote from the cap body communicating with the communication hole;
    A driving motor fixed to the mounting base, and electrically connected to said circuit control module, wherein said control circuit controls the driving operation of the motor assembly of;
    Drive guide rod fixed to the mounting base, the driving direction of the guide rod extending parallel to the axial direction of said hollow cylinder;
    The drive spindle, the drive motor is connected to one end of a transmission, said drive shaft extending direction of the lead screw is parallel to the hollow cylinder;
    A drive slider slidably sleeved on said drive rod and said drive rod guide, the driving of the slide remote from the push rod end of the piston is fixedly connected to the driving slide can be driven by said sliding pusher, the said piston sliding in said hollow cylinder.
  10. The apparatus according to claim 1, characterized in that, further comprising a fixedly mounted to the moving frame, for driving said fourth vision sensor assembly along said axis drive assembly slidably, said drive assembly comprising:
    Drive motor, secured to said mobile frame, and connected to the electrical control circuit assembly, said circuit control module controls operation of the drive motor;
    Two drive rollers, wherein said drive roller with a drive motor drivingly connected to the other of said drive roller rotatably mounted to the mobile frame;
    Drive belt, the two sleeved on the drive roller, and the drive is connected to two drive rollers;
    Drive screw, remote from the drive motor is fixedly connected to the drive roller, so that the drive roller rotatably driven by said drive screw, said drive screw extending along said fourth axis;
    Transmission guides, fixedly mounted to the mobile frame, drive the extending direction of the guide member parallel with the transmission screw;
    Drive slider, slidably disposed on the drive sleeve and the drive screw the guide member, wherein said visual sensor assembly is fixed to the drive slider.
  11. The apparatus according to claim 1, wherein said visual sensor assembly comprising:
    A fixing plate, along said fourth shaft slidably mounted to the mobile frame;
    A fixed housing, is fixedly connected to the fixing plate, and together with the fixing plate is formed with a receiving space having a gap through light;
    Vision sensor, received in the accommodation space, and fixedly connected with the fixed plate, and the visual sensor and the control circuit assembly electrically connected to said circuit control module controls operation of the visual sensor;
    Camera accommodated in the accommodating space, and fixed to said visual sensor, the camera and the visual sensor, the control circuit components are electrically connected to a camera for the sample collection vessel image, and the image of the camera can be transmitted to the acquired visual sensor, the vision sensor so that the image transmitted to said circuit control module;
    Mirror, fixedly mounted to the fixing plate, the position of the mirror with the light through the notch corresponding to the location, and positioned below the camera, so that the mirrors of the vessel the reflected image of the sample to the camera, the camera can acquire the image of the sample in the container.
  12. The apparatus according to claim 1, wherein the base comprises:
    Floor;
    Two guide plates, spaced and parallel relative to the base plate fixed on;
    Extending direction slide along the guide plate is slidably disposed between the two guide plates;
    Wherein said control circuit assembly, movement of the first shaft assembly and the drive assembly are fixed to the base plate, said container assembly mounted on the slide.
  13. The apparatus according to claim 12, characterized in that, for sensing the position of the mounting slide on the base plate, and is electrically connected to the control circuit assembly position sensor.
  14. The apparatus according to claim 12, wherein, wherein said container is a tube portion, another portion of the container is a supporting means for respectively separating the lymph fluid and the diluting liquid reagent bottle, the upper slide plate defines a receiving groove portion of the reagent bottle received in the receiving groove, said container assembly further comprises means for placing the test tube in test tube rack, the cuvette holder is detachably mounted on the slide.
  15. The apparatus according to claim 14, characterized in that the receiving groove is mounted for sensing the presence or absence of the reagent bottle, and the first status sensor electrically connected to said circuit control module; said slide plate It is also mounted a plurality of tubes for sensing the presence or absence, and the second state of the sensor assembly electrically connected to the control circuit.
  16. One kind of peripheral blood mononuclear cells in vitro extraction method, comprising:
    An in vitro to provide any one of the peripheral blood mononuclear cells 1 to 15, wherein the extraction means;
    The diluted fluid, lymph fluid and separating the three samples were loaded into a container;
    Said circuit control module controls said first shaft and said second shaft moving assembly moving at least one working assembly, so that the moving frame and the drive mechanism of the pipetting assembly to move the visual sensor, then the circuit control means for controlling the movement of the visual sensor assembly along said fourth axis slide frame, so that the vision sensor at a bearing assembly to move the container containing the sample, said control circuit controls said assembly vision sensor assembly acquiring images of the sample, the visual sensor assembly of the image transmitted to said circuit control module, said control circuit assembly of the sample is calculated according to the white layer of the image using the following formula position, wherein, assuming that P represents a point in space, defining the point P in the second coordinate axis and the coordinate system established as a third shaft (PAx, PAy), defining the point P the two-dimensional coordinate visual sensor coordinate system of the image acquisition assembly is located in the range (PCx, PCy), defining the original coordinate system of the image acquisition vision sensor assembly is located Point, and the coordinate origin of the second coordinate axis and the fourth axis to establish a coordinate system (OCx, OCy) defining the second axis of the fourth shaft to establish a second axis and the coordinate system is established in a third coordinate axis (OBx, OBy), R1, R2, R3 and R4 each represent a space between the rotation axis as transformation, scaling and translation factor:
    Figure PCTCN2016100293-appb-100001
    Said circuit controls the pipetting mechanism assembly along said third axis slide position control of the white film layer calculated, and the pipetting means extending into the vessel containing the sample up to the calculated the position of said white film layer, said circuit control means further controls the drive assembly to the pipetting means providing a suction force to cause the substance pipetting mechanism draw the white film of the sample;
    The control circuit controls the pipetting assembly means slidable along said third axis inverted, so that the pipetting means is removed from the vessel containing the sample, said circuit control means controls said first axis assembly and the second shaft moving in at least one working assembly, so that the moving frame driving the pipetting means slidably fitted to said container at a diluent, said circuit control means controls the pipetting mechanism in the third axis slide until the slide to a container containing the diluent, the circuit control module controls said drive assembly work, the pipetting means to send a force release of the substance albuginea injection layer containing said diluent vessel mixing, dilution to obtain a white film layer;
    The control circuit assembly for controlling the movement of said pipetting mechanism frame slides along the third axis, so that the pipetting means into said white film layer diluent, said circuit control means controls the drive assembly to the pipetting means providing a suction force to cause the substance pipetting mechanism draw the diluent white film layer;
    Said circuit control module controls said first shaft and said second shaft moving assembly moving at least one working assembly, to drive the movement of the slide frame to move the pipetting means to the bearing containing the isolated lymph the liquid container, and said circuit control means controls the pipetting mechanism of the mobile frame along the third axis, and controlling the drive of the pipetting assembly to provide a means to force a release of a substance the buffy coat was diluted into the container with the bearing lymph separated liquid, and finally by centrifugation to obtain peripheral blood mononuclear cells.
  17. The method according to claim 16, wherein said suction mechanism pipetting step of the sample in the white film layer, said pipetting means to draw concentric circles moving said white film layer .
  18. The method according to claim 16, wherein the step to draw the sample in the white film in the pipetting means, a pipetting means movement along an Archimedes spiral to draw the said white film layer.
  19. The method according to claim 16, wherein said suction mechanism pipetting step of the sample in the white film layer, said Z-shaped pipetting mechanism made movable to draw said white film layer .
  20. The method according to claim 16, wherein said control circuit assembly for controlling the movement of said pipetting mechanism frame along the third axis, and to control the drive assembly to the pipetting means the step of providing a release force to said substance white film layer containing said diluent injection bearing lymph liquid separation vessel is: the circuit control module controls the pipetting mechanism moves along the frame axis of said third means until said pipetting a liquid from the high end of the 0.5 ~ 1mm lymph liquid separation liquid level, said circuit control means controls said drive assembly to the pipetting means provided a force release rate of the substance to the white film layer dilution was 0.01ml / s ~ 0.3m / s is injected into the vessel containing the bearing lymph separated liquid in the diluent so that the white film layer floating on the surface of lymphoid separated liquid, until the buffy coat in a diluent injecting 3 ~ 5ml, and then to 0.3ml / s 0.8m / s velocity of the injection - the remaining buffy coat dilution .
PCT/CN2016/100293 2016-09-27 2016-09-27 In vitro extraction device and method for peripheral blood mononuclear cells WO2018058286A1 (en)

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WO2007041716A1 (en) * 2005-10-05 2007-04-12 Gambro Bct, Inc. Method and apparatus for leukoreduction of red blood cells
CN102286360A (en) * 2011-06-21 2011-12-21 中国人民解放军第四军医大学 An apparatus for the separation of peripheral blood mononuclear cells for
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