CN111500426B - Micro-operation suction device for sorting porcine oocytes in mixed solution - Google Patents
Micro-operation suction device for sorting porcine oocytes in mixed solution Download PDFInfo
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- CN111500426B CN111500426B CN202010181340.4A CN202010181340A CN111500426B CN 111500426 B CN111500426 B CN 111500426B CN 202010181340 A CN202010181340 A CN 202010181340A CN 111500426 B CN111500426 B CN 111500426B
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- needle head
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- culture dish
- lens
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
Abstract
The invention provides a micro-operation suction device for sorting porcine oocytes in a mixed solution, which comprises a supporting mechanism, a power mechanism, a suction mechanism, a needle head moving mechanism, a culture dish moving mechanism and a visual system, wherein the supporting mechanism is fixed on a two-dimensional operating platform, and the power mechanism is driven by external force to provide suction power for the suction mechanism; the suction mechanism adopts the syringe piston principle to suck the cells; the needle head moving mechanism is arranged to move up and down to adjust the position of the needle head; the visual system comprises a visual lens and an image processing module, wherein the visual lens is arranged to move up and down to adjust the position, the pig egg cells and the culture solution in the culture dish are photographed, and then the analysis processing is carried out by the image processing module; the culture dish moving mechanism is set to be in a rotatable moving mode so as to realize that the culture dish is randomly switched between the needle head and the lens position, and the control system controls the two-dimensional operation platform to translate and find the pig oocyte according to the visual information fed back by the image processing module and sucks the pig oocyte through the needle head.
Description
Technical Field
The invention belongs to the technical field of micro-operation, and particularly relates to a suction device for sorting porcine oocytes in a mixed solution.
Background
With the development of scientific technology, scientific research and clinical application in the fields of life science and the like tend to be more miniaturized and refined, wherein the micro-operation technology of cells is a technical support for promoting the development trend. Whether the relevant scientific research and clinical application such as gene engineering, cloning technology, biological medicine research and development can make breakthrough progress is also closely related to the micro-operation technology of cells.
At present, the cell micro-operation technology still depends on manual operation, and the requirement on the richness of the manual experience is higher. In the manual micro-operation sorting process of the pig oocytes, the operator is required to concentrate on the attention for a long time, the speed and the quantity of the cells sucked by the operator are required to be accurately controlled, the small hand shaking of the operator can damage the pig oocytes, and therefore the related workers are required to have abundant operation experience and high operation precision, otherwise the activity of the pig oocytes can be influenced.
In summary, it is a problem to be solved that how to provide a micro-manipulation suction device to reduce the damage caused by the sorting process of the oocytes of the manual micro-manipulation pig and reduce the dependence of the manual manipulation.
Disclosure of Invention
In order to overcome a series of defects in the prior art, the invention aims to provide a micro-operation suction device for sorting porcine oocytes in a mixed solution, which is characterized by comprising a supporting mechanism, a power mechanism, a suction mechanism, a needle head moving mechanism, a culture dish moving mechanism and a visual system, wherein the power mechanism provides power for suction for the suction mechanism through external force driving; the suction mechanism adopts the syringe piston principle to suck the cells; the needle moving mechanism is arranged to move up and down to adjust the position of the needle 7; the vision system comprises a vision lens 21 and an image processing module, wherein the vision lens 21 is arranged to move up and down to adjust the position, the pig egg cells and the culture solution in the culture dish 25 are photographed, and then the analysis processing is carried out by the image processing module; the culture dish moving mechanism is arranged in a rotatable moving mode so as to realize that the culture dish is switched at the positions of the needle head and the lens at will.
Further, the supporting mechanism comprises a base 1 and a support 2, the base 1 is fixed on the two-dimensional operating platform, and the support 2 is fixed on the base 1.
Further, syringe needle moving mechanism includes differential head support 3, differential head 4, syringe needle installation clamp 6 and syringe needle 7, and differential head support 3 fixes on support 2, and differential head 4 is fixed the bottom of differential head support 3, the lower extreme of differential head 4 is fixed with syringe needle installation clamp 6, installs syringe needle 7 on the syringe needle installation clamp 6, reciprocates in the vertical direction through differential head 4 to drive syringe needle 7 and reciprocate.
Furthermore, the power mechanism comprises a bracket connecting piece 8, a bearing 9, a bearing seat 10, a handle 11, a gear 12 and a rack 13, wherein the bracket connecting piece 8 is fixed on the bracket 2 through a screw, the bearing seat 10 is connected on the bracket connecting piece 8, the bearing 9 is arranged on the bearing seat 10, two ends of the bearing 9 are respectively connected with the handle 11 and the gear 12, and the gear 12 and the rack 13 form meshing transmission.
Further, suction mechanism includes cylinder connecting piece 5, syringe needle 7, push rod 14, cylinder 15, hose 16 and rubber cap, and cylinder connecting piece 5 is connected in differential head 4, is fixed with cylinder 15 on the cylinder connecting piece 5, and cylinder 15 links together through hose 16 with syringe needle 7 on the syringe needle mounting clip 6, and sliding connection has the lower part to cup joint rubber cap's push rod 14 in cylinder 15, the upper portion of push rod 14 is connected with rack 13, through external force drive handle 11, drives push rod 14 and reciprocates, provides the power of suction or discharge usefulness for push rod 14.
Furthermore, the same side of the needle head mounting clamp 6 and the needle cylinder connecting piece 5 is fixedly connected with the same positioning column which is vertically arranged.
Further, the vision system further comprises a lens support 23, the lens support 23 is fixed on the support 2, the vision lens 21 is connected to the lens support 23, and the lens support 23 adjusts the height and the position of the vision lens 21 through a knob.
Further, the culture dish moving mechanism is arranged in the following manner: a rotary platform 20 is arranged below the support 2, a culture dish 25 for containing pig oocytes and culture solution thereof is arranged on the rotary platform 20, the rotary platform 20 is installed on a steering engine 17, the steering engine 17 is fixed on a steering engine support 18 through screws, the steering engine support 18 is fixed on a steering engine support 19, and the culture dish 25 is slowly moved to the position right below the visual lens 21 or the position right below the needle head 7 through the rotation of the rotary platform 20.
Further, the image processing module adopts VS2015+ OpenCV software to process the image of the culture dish 25 shot by the vision lens 21, and obtains the position of the pig oocyte in the culture dish 25 and feeds the position back to the control system;
and the control system controls the two-dimensional operation platform to move in a plane and find the pig oocyte according to the visual information fed back by the image processing module, and then the pig oocyte is sucked by the needle head 7.
Furthermore, the pig oocytes are intermittently sucked by the up-and-down movement of the needle 7 in the vertical direction.
Compared with the prior art, the invention provides a micro-operation suction device for sorting porcine oocytes in a mixed solution, which has the following beneficial effects:
1) the invention reduces the dependence degree on manpower and improves the operation efficiency;
2) the visual lens and the image processing module are added, so that the cells are well identified, the operation time is reduced, and the artificial visual fatigue is relieved;
3) the invention has compact mechanical structure, low part processing cost, high space utilization rate and strong operability;
4) the invention uses screw connection, which is convenient for assembly and disassembly and replacement of part of parts;
5) the micro-operation object can be replaced at any time according to actual needs, and the adaptability is wide.
Drawings
FIG. 1 is a schematic overall view of a micro-manipulation pipette device for sorting porcine oocytes in a mixed solution according to the present invention;
FIG. 2 is a front view of a micro-manipulation pipette device for sorting porcine oocytes in a mixed solution according to the present invention;
FIG. 3 is a top view of a needle mounting clip of a micro-manipulation pipette device for sorting porcine oocytes in a mixed solution according to the present invention;
FIG. 4 is an overall schematic view of a needle mounting clip of a micro-manipulation pipette device for sorting porcine oocytes in a mixed solution according to the present invention;
FIG. 5 is a schematic view of a gear and rack connection structure of a micro-manipulation suction device for sorting porcine oocytes in a mixed solution according to the present invention;
the reference numbers in the figures are:
1-base, 2-bracket, 3-differential head bracket, 4-differential head, 5-syringe connecting piece, 6-syringe mounting clamp, 7-syringe, 8-bracket connecting piece, 9-bearing, 10-bearing seat, 11-handle, 12-gear, 13-rack, 14-push rod, 15-syringe, 16-hose, 17-steering engine, 18-steering engine bracket, 19-steering engine support, 20-rotary platform, 21-vision lens, 23-lens bracket and 25-culture dish.
Detailed Description
In order to make the implementation objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be described in more detail below with reference to the accompanying drawings in the embodiments of the present invention. In the drawings, the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The described embodiments are only some, but not all embodiments of the invention.
All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiments and the directional terms described below with reference to the drawings are exemplary and intended to be used in the explanation of the invention, and should not be construed as limiting the invention.
In one broad embodiment of the invention, a micro-manipulation pipette device for sorting porcine oocytes in a mixed solution includes a support mechanism, a power mechanism, a suction mechanism, a needle movement mechanism, a culture dish movement mechanism, and a vision system. The supporting mechanism is used for fixedly connecting the power mechanism, the needle head moving mechanism and the visual system; the power mechanism is fixedly connected to the supporting mechanism and provides power for suction for the suction mechanism through external force driving; the suction mechanism is fixedly connected on the power mechanism, and realizes the cell suction function by adopting the syringe piston principle; the needle head moving mechanism is also fixedly connected to the supporting mechanism and is set to be in an adjustable mode of moving up and down so as to adjust the position of the needle head 7; the culture dish moving mechanism is arranged in a rotatable moving mode so as to realize the random switching of the culture dish 25 between the positions of the needle head 7 and the visual lens 21; the vision system is also fixedly connected to the supporting mechanism and comprises a vision lens 21 and an image processing module, wherein the vision lens 21 is set to be in an adjustable mode of moving up and down and is used for photographing the egg cells and the culture solution in the culture dish 25 and then carrying out analysis processing by the image processing module.
Preferred embodiments of the present invention will be described in detail below with reference to fig. 1 to 5.
Preferably, the supporting mechanism comprises a base 1 and a support 2, the base 1 is fixed on the two-dimensional operating platform, the support 2 is fixed on the base 1, and the supporting mechanism is kept static relative to the two-dimensional operating platform.
Preferably, the needle moving mechanism comprises a differential head support 3, a differential head 4, a needle mounting clamp 6 and a needle 7, the differential head support 3 is fixed on the support 2, the differential head 4 is fixed at the bottom of the differential head support 3, the needle mounting clamp 6 is fixed at the lower end of the differential head 4, the needle 7 is mounted on the needle mounting clamp 6, and the needle moves up and down in the vertical direction through the differential head 4 and drives the needle 7 to move up and down.
Preferably, the power mechanism comprises a bracket connecting piece 8, a bearing 9, a bearing seat 10, a handle 11, a gear 12 and a rack 13, the bracket connecting piece 8 is fixed on the bracket 2 through a screw, the bearing seat 10 is connected on the bracket connecting piece 8, the bearing 9 is arranged on the bearing seat 10, two ends of the bearing 9 are respectively connected with the handle 11 and the gear 12, and the gear 12 and the rack 13 form meshing transmission.
Preferably, the suction mechanism comprises a syringe connecting piece 5, a needle 7, a push rod 14, a syringe 15, a hose 16 and a rubber cap, the syringe connecting piece 5 is connected to the differential head 4, the syringe 15 and the differential head 4 can be fixed through the syringe connecting piece 5, a certain relative position relation is ensured between the syringe 15 and the differential head 4, the syringe 15 is fixed on the syringe connecting piece 5, the syringe 15 and the needle 7 on the needle mounting clamp 6 are connected together through the hose 16, the push rod 14 with the lower part sleeved with the rubber cap is connected in the syringe 15 in a sliding manner, the upper part of the push rod 14 is connected with a rack 13, and the handle 11 is driven by external force to drive the push rod 14 to move up and down, so that power for suction or discharge is provided for the push rod 14.
Preferably, the same side fixedly connected with reference column of same vertical setting of syringe needle installation clamp 6 and cylinder connecting piece 5 has further guaranteed that syringe needle 7 can only reciprocate on vertical direction.
Preferably, the vision system further comprises a lens support 23, the lens support 23 is fixed on the support 2, the vision lens 21 is connected to the lens support 23, and the lens support 23 adjusts the height and the position of the vision lens 21 through a knob.
Preferably, the culture dish moving mechanism is arranged in the following manner: a rotary platform 20 is arranged below the support 2, a culture dish 25 for containing pig oocytes and culture solution thereof is arranged on the rotary platform 20, the rotary platform 20 is installed on a steering engine 17, the steering engine 17 is fixed on a steering engine support 18 through screws, the steering engine support 18 is fixed on a steering engine support 19, and the culture dish 25 is slowly moved to the position right below the visual lens 21 or the position right below the needle head 7 through the rotation of the rotary platform 20.
Preferably, the image processing module adopts VS2015+ OpenCV software to process the image of the culture dish 25 taken by the vision lens 21, and obtains the position of the pig oocyte in the culture dish 25 and feeds the position back to the control system;
and the control system controls the two-dimensional operation platform to move in a plane and find the pig oocyte according to the visual information fed back by the image processing module, and then the pig oocyte is sucked by the needle head 7.
Preferably, the porcine oocytes are intermittently aspirated by moving the needle 7 up and down in the vertical direction.
After the image processing module finds the porcine oocytes needing to be grabbed, the steering engine 17 is used for driving the rotary platform 20 to slowly rotate to the position below the needle head 7, the needle head 7 is positioned to the position above the porcine oocytes needing to be extracted by operating the two-dimensional platform, the required porcine oocytes are screened out by using the needle head 7, in addition, for the convenience of observation and counting, the porcine oocytes and a culture solution thereof need to be divided into a plurality of sections for suction in the hose 16, the solution is that when the interruption occurs, the needle head 7 is just drawn out from the liquid level to suck a section of air, and the intermittent extraction can be realized.
Of particular note are:
firstly, the image processing of the invention is based on a Visual Studio2015 software development platform and an OpenCV Visual database, and an image recognition program is compiled and operated to realize the recognition and positioning process of a target object by configuring environment variables;
secondly, the two-dimensional operation platform, the control system and the parts which are not involved are the same as or can be realized by adopting the prior art, and the device has reasonable design structure and convenient use and can meet the use requirements of people.
Finally, it should be pointed out that: the above examples are only for illustrating the technical solutions of the present invention, and are not limited thereto. Although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (6)
1. A micro-operation suction device for sorting porcine oocytes in a mixed solution is characterized by comprising a supporting mechanism, a power mechanism, a suction mechanism, a needle head moving mechanism, a culture dish moving mechanism and a visual system, wherein the power mechanism provides power for suction for the suction mechanism through external force driving; the suction mechanism adopts the syringe piston principle to suck the cells; the needle head moving mechanism is arranged to move up and down to adjust the position of the needle head (7); the visual system comprises a visual lens (21) and an image processing module, wherein the visual lens (21) is arranged to move up and down to adjust the position, the pig egg cells and the culture solution in the culture dish (25) are photographed, and then the analysis processing is carried out by the image processing module; the culture dish moving mechanism is arranged in a rotatable moving mode so as to realize the random switching of the culture dish at the positions of the needle head and the lens;
the supporting mechanism comprises a base (1) and a support (2), the base (1) is fixed on the two-dimensional operating platform, and the support (2) is fixed on the base (1);
the needle head moving mechanism comprises a differential head support (3), a differential head (4), a needle head mounting clamp (6) and a needle head (7), the differential head support (3) is fixed on the support (2), the differential head (4) is fixed at the bottom of the differential head support (3), the needle head mounting clamp (6) is fixed at the lower end of the differential head (4), the needle head (7) is mounted on the needle head mounting clamp (6), and the needle head moving mechanism moves up and down in the vertical direction through the differential head (4) and drives the needle head (7) to move up and down;
the power mechanism comprises a support connecting piece (8), a bearing (9), a bearing seat (10), a handle (11), a gear (12) and a rack (13), the support connecting piece (8) is fixed on the support (2) through a screw, the bearing seat (10) is connected onto the support connecting piece (8), the bearing (9) is placed on the bearing seat (10), the two ends of the bearing (9) are respectively connected with the handle (11) and the gear (12), and the gear (12) and the rack (13) form meshing transmission;
suction mechanism includes cylinder connecting piece (5), syringe needle (7), push rod (14), cylinder (15), hose (16) and rubber cap, and cylinder connecting piece (5) are connected in differential head (4), is fixed with cylinder (15) on cylinder connecting piece (5), and cylinder (15) link together through hose (16) with syringe needle (7) on syringe needle mounting clip (6), and sliding connection has lower part to cup joint push rod (14) of rubber cap in cylinder (15), the upper portion of push rod (14) is connected with rack (13), through external force drive handle (11), drives push rod (14) and reciprocates, provides the power of suction or discharge usefulness for push rod (14).
2. A micro-operation suction device according to claim 1, wherein the same side of the needle mounting clip (6) and the syringe connecting member (5) is fixedly connected with the same positioning column which is vertically arranged.
3. The micro-operation suction device according to claim 1, wherein the vision system further comprises a lens holder (23), the lens holder (23) is fixed on the holder (2), the vision lens (21) is connected to the lens holder (23), and the lens holder (23) adjusts the height and the position of the vision lens (21) through a knob.
4. The micro-manipulated pipetting device of claim 1, wherein the plate movement mechanism is configured to: a rotary platform (20) is arranged below the support (2), a culture dish (25) for containing pig oocytes and culture solution of the pig oocytes is arranged on the rotary platform (20), the rotary platform (20) is installed on a steering engine (17), the steering engine (17) is fixed on a steering engine support (18) through screws, the steering engine support (18) is fixed on a steering engine support (19), and the culture dish (25) is slowly moved to the position under the visual lens (21) or the position under the needle head (7) through rotation of the rotary platform (20).
5. The micro-manipulated suction device of claim 1,
the image processing module adopts VS2015+ OpenCV software to process images of the culture dish (25) shot by the visual lens (21), and obtains the position of the pig oocyte in the culture dish (25) and feeds the position back to the control system;
the control system controls the two-dimensional operation platform to move in a plane and find the porcine oocytes according to the visual information fed back by the image processing module, and then the porcine oocytes are sucked through the needle head (7).
6. The micromanipulation pipetting device of claim 5, wherein the porcine oocytes are intermittently pipetted by moving the needle (7) up and down in the vertical direction.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010181340.4A CN111500426B (en) | 2020-03-16 | 2020-03-16 | Micro-operation suction device for sorting porcine oocytes in mixed solution |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010181340.4A CN111500426B (en) | 2020-03-16 | 2020-03-16 | Micro-operation suction device for sorting porcine oocytes in mixed solution |
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| CN111500426A CN111500426A (en) | 2020-08-07 |
| CN111500426B true CN111500426B (en) | 2022-03-01 |
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Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3718066A1 (en) * | 1987-05-29 | 1988-12-08 | Zeiss Carl Fa | METHOD FOR MICROINJECTION IN CELLS OR. FOR SUCTION FROM SINGLE CELLS OR WHOLE CELLS FROM CELL CULTURES |
| US7776584B2 (en) * | 2003-08-01 | 2010-08-17 | Genetix Limited | Animal cell colony picking apparatus and method |
| JP4948481B2 (en) * | 2008-06-27 | 2012-06-06 | オリンパス株式会社 | Cell manipulation observation device |
| EP2423661A1 (en) * | 2010-08-30 | 2012-02-29 | Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt GmbH | Device and method for automatic isolation and transfer of at least one microscopic sample from a sample holder to a holding system |
| CA2812776C (en) * | 2010-09-27 | 2022-05-17 | Auxogyn, Inc. | Apparatus, method, and system for the automated imaging and evaluation of embryos, oocytes, and stem cells |
| CN102321530B (en) * | 2011-09-08 | 2013-04-17 | 上海炎景生物工程有限公司 | Pick-up head for automatic colony picking device |
| EP2933327A4 (en) * | 2012-12-12 | 2016-08-03 | Hitachi Chemical Co Ltd | Cancer cell isolation device and cancer cell isolation method |
| CN106350437B (en) * | 2016-06-13 | 2019-10-15 | 暨南大学 | A kind of cell ball positioning biometric print device and method |
| CN106381259B (en) * | 2016-09-27 | 2018-09-11 | 深圳市港科深研生物科技有限公司 | The vitro extraction device of peripheral blood mononuclear cells and the vitro extraction method of peripheral blood mononuclear cells |
| CN110607229A (en) * | 2019-09-29 | 2019-12-24 | 深圳赛动生物自动化有限公司 | Cell transfer observation mechanism and working method thereof |
| CN111004717A (en) * | 2019-12-26 | 2020-04-14 | 常州市武进人民医院 | Single cell separation and collection device |
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