CN108070618A - A kind of method of microinjection operation - Google Patents
A kind of method of microinjection operation Download PDFInfo
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- CN108070618A CN108070618A CN201711315349.4A CN201711315349A CN108070618A CN 108070618 A CN108070618 A CN 108070618A CN 201711315349 A CN201711315349 A CN 201711315349A CN 108070618 A CN108070618 A CN 108070618A
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- needle
- injection needle
- fixed pin
- motion arm
- injection
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/89—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection
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Abstract
The invention discloses a kind of methods of microinjection operation, including at least drawing needle apparatus, fixed pin, injection needle, fixed pin motion arm, injection needle motion arm and microscope, the fixed pin is rotatablely connected with fixed pin motion arm, the injection needle is rotatablely connected with injection needle motion arm, the microscope includes objective table, objective table is equipped with cell carrier, the fixed pin motion arm is equipped with the first knob, the injection needle motion arm is equipped with the second knob, and the fixed pin has 30 ° of bending angle.The angle of a kind of position by adjusting injection needle and fixed pin is provided, a kind of conveniently a kind of method of microinjection operation of the pin mouth of suitable size can be conveniently obtained in injection needle broken needle.
Description
Technical field
The present invention relates to microinjection operations, and in particular to a kind of method of microinjection operation.
Background technology
Microinjection technique be will be injected in field of microscope cell adjustment it is fixed after, using microinjection pin by external source
A kind of technology of gene or the injection of other substances into the cell, is one of important technical of modern biological project, especially turns
The cell engineerings such as gene, clone and test-tube baby field.Compared with other transgenic technologys, the advantages of microinjection is transgenosis
Stabilised efficiency, injection gene size be unrestricted, without carrier and operation object without particular/special requirement etc., protected in animals on the brink of extinction
Shield, reproduction in health, species improvement and intracellular caryoplasm research with being used widely.
When microinjection operates, a step of most critical is exactly the broken needle of injection needle.The needle point fracture of injection needle is excessively thick, injection
When cell the death rate increase；And fracture is meticulous, the amount of injection can be very few and easily blocks up pin.The general broken needle method of injection needle
It is that injection needle hits fixed needle wall to sever injection needle needle point, but the size of pin mouth is random after broken needle, and after broken needle
Pin mouth internal diameter needs suitable size, so this operation needs certain technology content and the operating time is generally longer also
With contingency.Requirement of this method to injection needle is also very high, and injection needle needle point will very sharp very short certain hardness again.It draws
Because of parameter and machine difference when needle apparatus draws injection needle, what is pulled out is diversified in specifications, and the injection needle needle point pulled out is most of all
Certain length and flexible are had, above-mentioned method is with regard to this injection needle of uncomfortable make and break.
China Patent No. CN105420099A, publication date be on March 23rd, 2016, entitled efficient microinjection
Device, including quartz substrate and injection body, injection body is fixed on quartz substrate, and the injection body is micro- equipped with inputting
Runner, output fluid channel and cell delivery fluid channel are injected and also set up on body between input fluidic channels and output fluid channel
There are microinjection runner and control cell movement runner, microinjection runner is vertical direction and vertical with cell delivery fluid channel
It connects, is fixed with injection needle on microinjection runner, there is gap, and gap between microinjection runner and injection needle
Size is less than cell dia, has limit protrusion in microinjection runner, and control cell movement runner hangs down with microinjection runner
It is direct-connected logical, and cell movement runner is controlled to be located above limit protrusion, a kind of movement by controlling microfluid is provided come between
Connect control
Cell movement makes cell " oneself " knock injection needle, so as to complete one kind of the microinjection of cell operation efficiently
Microinjection device.It is disadvantageous in that, it is impossible to which the corresponding injection needle needle point that needle apparatus is drawn to pull out controls the interior of broken needle pin mouth
Footpath.
The content of the invention
The present invention controls the internal diameter of broken needle pin mouth for the above-mentioned injection needle needle point that cannot correspond to drawing needle apparatus pull-out of solution
Problem provides the angle of a kind of position by adjusting injection needle and fixed pin, can be conveniently in injection needle broken needle
Obtain a kind of conveniently a kind of method of microinjection operation of the pin mouth of suitable size.
The present invention is that technical solution is used by solving its technical problem：A kind of method of microinjection operation, at least
Including drawing needle apparatus, fixed pin, injection needle, fixed pin motion arm, injection needle motion arm and microscope, the fixed pin with
Fixed pin motion arm is rotatablely connected, and the injection needle is rotatablely connected with injection needle motion arm, and the microscope includes loading
Platform, objective table are equipped with cell carrier, and the fixed pin motion arm is equipped with the first knob, on the injection needle motion arm
Equipped with the second knob, the fixed pin has 30 ° of bending angle.
Preferably, the present invention comprises the following steps：
Step 1：Cell is placed in the cell carrier on microscope carrier；
Step 2：Injection needle motion arm is adjusted by the first knob, the needle point of injection needle is made entirely to be laid in the bottom of cell carrier
Portion；
Step 3：Fixed pin is rotated, makes the curved back of rotation pin upward；
Step 4：Fixed pin motion arm is adjusted by the second knob, fixed pin syringe needle is made slowly to be contacted with vessel bottom；
Step 5：Fixed pin motion arm is adjusted by the second knob while adjusting injection needle motion arm by the first knob, is made
Injection needle needle point and fixed pin syringe needle are on same horizontal plane；
Step 6：The position of injection needle needle point is selected, fixed pin motion arm is finely tuned by the second knob；
Step 7：Injection needle needle point is broken with fixed pin syringe needle, broken needle is completed.
Preferably, the internal diameter of pin mouth is 3-5 μm after the broken needle.Pin mouth internal diameter is for 3-5 μm of size after broken needle
Most suitable, the needle point fracture of injection needle is excessively thick, and the death rate of cell increases during injection；And fracture is meticulous, the amount of injection can mistake
Block up less and easily pin.
Preferably, the injection needle length of needlepoint is 5-8mm.The needle point of injection needle of the present invention has stronger hardness,
Needle point has high sharpness.
Preferably, the length of the injection needle needle point that needle apparatus is drawn to pull out is 10-20mm.
The present invention practical significance be：
（1）It is convenient and efficient and easy to operate；
（2）By adjusting the position of injection needle and the angle of fixed pin, it is suitable easily and efficiently to be obtained in injection needle broken needle
The pin mouth of size.
（3）Because of parameter and machine difference when being suitable for drawing needle apparatus drawing injection needle, the injection diversified in specifications pulled out
Pin.
Specific embodiment
When microinjection operates, a step of most critical is exactly the broken needle of injection needle.The needle point fracture of injection needle is excessively thick, injection
When cell the death rate increase；And fracture is meticulous, the amount of injection can be very few and easily blocks up pin.The general broken needle method of injection needle
It is that injection needle hits fixed needle wall to sever injection needle needle point, but the size of pin mouth is random after broken needle, and after broken needle
Pin mouth internal diameter is just most suitable for 3-5 μm of size, so this operation needs certain technology content and the operating time is general
It is longer that also there is contingency.Requirement of this method to injection needle is also very high, and injection needle needle point is very sharp very short（5-8mm）Again
Certain hardness.Because of parameter and machine difference when drawing needle apparatus drawing injection needle, what is pulled out is diversified in specifications, and the injection needle pulled out
Needle point largely can all have certain length（10-20mm）And flexible, the present embodiment by adjusting injection needle position and
The angle of fixed pin can easily and efficiently obtain the pin mouth of suitable size in injection needle broken needle.
The present embodiment includes loading on the motion arm and microscope carrier of fixed pin, injection needle, fixed pin and injection needle
The vessel of cell.Injection needle needle point has certain length and toughness, and slowly being adjusted by the motion arm of injection needle makes injection needle pin
It is sharp to be entirely laid in the vessel bottom for loading cell.Fixed pin adjusts its angle when being attached on motion arm（Fixed pin has
30 ° or so of bending angle）Make its curved back upward, adjust fixed pin motion arm make fixed pin syringe needle slowly with vessel
Bottom contacts.Then adjusting two motion arms makes injection needle needle point and fixed pin syringe needle select injecting on a horizontal plane
Fixed pin motion arm is finely tuned in the position of the suitable size of pin needle point, breaks injection needle needle point with fixed pin syringe needle, broken needle is completed.
Embodiment described above is a kind of preferred version of the present invention, not makees limit in any form to invention
System also has other variants and remodeling on the premise of without departing from the technical solution recorded in claim.
Claims (5)
- A kind of 1. method of microinjection operation, which is characterized in that including at least drawing needle apparatus, fixed pin, injection needle, fixed pin Motion arm, injection needle motion arm and microscope, the fixed pin are rotatablely connected with fixed pin motion arm, the injection needle It is rotatablely connected with injection needle motion arm, the microscope includes objective table, and objective table is equipped with cell carrier, the fixation Pin motion arm is equipped with the first knob, and the injection needle motion arm is equipped with the second knob, and the fixed pin has 30 ° curved Bent angle.
- A kind of 2. method of microinjection operation, which is characterized in that comprise the following steps：Step 1：Cell is placed in the cell carrier on microscope carrier；Step 2：Injection needle motion arm is adjusted by the first knob, the needle point of injection needle is made entirely to be laid in the bottom of cell carrier Portion；Step 3：Fixed pin is rotated, makes the curved back of rotation pin upward；Step 4：Fixed pin motion arm is adjusted by the second knob, fixed pin syringe needle is made slowly to be contacted with vessel bottom；Step 5：Fixed pin motion arm is adjusted by the second knob while adjusting injection needle motion arm by the first knob, is made Injection needle needle point and fixed pin syringe needle are on same horizontal plane；Step 6：The position of injection needle needle point is selected, fixed pin motion arm is finely tuned by the second knob；Step 7：Injection needle needle point is broken with fixed pin syringe needle, broken needle is completed.
- A kind of 3. method of microinjection operation according to claim 2, which is characterized in that pin mouth after the broken needle Internal diameter is 3-5 μm.
- 4. the method for a kind of microinjection operation according to claim 1, which is characterized in that the injection needle needle point is long It spends for 5-8mm.
- 5. the method for a kind of microinjection operation according to claim 1, which is characterized in that the drawing needle apparatus pulls out Injection needle needle point length be 10-20mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711315349.4A CN108070618B (en) | 2017-12-12 | 2017-12-12 | Microinjection operation method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711315349.4A CN108070618B (en) | 2017-12-12 | 2017-12-12 | Microinjection operation method |
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| CN108070618A true CN108070618A (en) | 2018-05-25 |
| CN108070618B CN108070618B (en) | 2021-03-05 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402220A (en) * | 2018-11-28 | 2019-03-01 | 安徽农业大学 | A method of collecting blastocoele liquid |
| CN111693196A (en) * | 2020-06-16 | 2020-09-22 | 梁诗豪 | Live embryo surface zona pellucida tension detection device |
| CN111909839A (en) * | 2019-05-08 | 2020-11-10 | 香港城市大学深圳研究院 | System for delivering substances into cells, master control device, microcontroller and method |
| CN112779119A (en) * | 2019-11-08 | 2021-05-11 | 中国科学院大连化学物理研究所 | Small manual five-dimensional needle tip adjusting frame for cell experiment |
| WO2023068924A1 (en) * | 2021-10-22 | 2023-04-27 | Life Science Methods B.V. | Microinjection system and method |
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| CN107245431A (en) * | 2017-08-04 | 2017-10-13 | 重庆三峡医药高等专科学校 | A kind of microinjection device precisely injected for cell drug and its operating method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402220A (en) * | 2018-11-28 | 2019-03-01 | 安徽农业大学 | A method of collecting blastocoele liquid |
| CN111909839A (en) * | 2019-05-08 | 2020-11-10 | 香港城市大学深圳研究院 | System for delivering substances into cells, master control device, microcontroller and method |
| CN112779119A (en) * | 2019-11-08 | 2021-05-11 | 中国科学院大连化学物理研究所 | Small manual five-dimensional needle tip adjusting frame for cell experiment |
| CN111693196A (en) * | 2020-06-16 | 2020-09-22 | 梁诗豪 | Live embryo surface zona pellucida tension detection device |
| WO2023068924A1 (en) * | 2021-10-22 | 2023-04-27 | Life Science Methods B.V. | Microinjection system and method |
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| CN108070618B (en) | 2021-03-05 |
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