CN207793228U - A kind of novel gene injection needle - Google Patents

Abstract

The utility model belongs to field of transgenic technology more particularly to a kind of novel gene injection needle.The shape of the injection needle and inner space are in coniform, the tip internal diameter of the injection needle is 0.5 μm, internal diameter of the injection needle at 30 μm away from the tip is 10 μm, internal diameter of the injection needle at 50 μm away from the tip is 20 μm, and volume of the injection needle at 45 μm away from the tip is 3pL.The injection needle is simple in structure, reasonable setting is carried out to height, internal diameter, volume etc., thus when carrying out micromanipulation, the foreign gene that corresponding amount can be drawn according to the position of injection needle high scale is injected, injection dosage can be accurately controlled, injection dosage pL class precisions are can reach.According to the scale sucking foreign gene marked on injection needle, it ensure that pL class precisions are not influenced by gene fluid viscosity, the injection dosage for avoiding human factor introducing differs and causes embryonic development after injecting inconsistent.

Description

A kind of novel gene injection needle

Technical field

The utility model belongs to field of transgenic technology more particularly to a kind of novel gene injection needle.

Background technology

Transgenic technology is the widely applied technology of life science, and microinjection is current structure transgenic animals The main method of model.The allogenic gene microinjection of animal embryo is a step of extremely key, and the size of injection dosage is closed It is that the survival rate of vitro Development of Embryos and the integration effect of allogenic gene are directly influenced to the foreign gene amount of holding to cell Fruit influences the success rate of transgenic animals.The size influence factor of injection dosage mainly have injection pressure, gene fluid viscosity, The shape of injection needle and end internal diameter etc..

Currently, mainly using hydraulic pressure manual mode microinjector with using compressed gas as the microinjection amount controller of medium into Row microinjection.Hydraulic pressure manual mode microinjector cannot accurately control injection dosage, it is difficult to ensure that pL (picoliter) class precision, The injection dosage that human factor introduces differs and causes embryonic development after injecting inconsistent.Using compressed gas as the microinjection of medium Though amount controller more can consistently control injection dosage, the instantaneous pressure generated in injection process can be to inside embryo knot It is configured to mechanical damage, duplicate injection error is big, and shape and end internal diameter pair according to injection needle are required for when per injection Injection pressure is adjusted with the time, furthermore the microinjection amount controller is expensive, and the economic cost paid is larger.

Utility model content

(1) technical problems to be solved

The technical problem to be solved by the present invention is to provide a kind of simple in structure and can accurately control injection dosage Novel gene injection needle.

(2) technical solution

In order to solve the above-mentioned technical problem, the utility model provides a kind of novel gene injection needle, the note It is in coniform to penetrate the shape of needle and inner space, and the tip internal diameter of the injection needle is 0.5 μm, and the injection needle is away from described Internal diameter at 30 μm of tip is 10 μm, and internal diameter of the injection needle at 50 μm away from the tip is 20 μm, and the injection needle exists Volume at 45 μm away from the tip is 3pL.

Further, it is equipped with label on outer wall of the injection needle at 45 μm away from the tip.

Further, it is equipped with label on outer wall of the injection needle at 50 μm away from the tip.

Further, it is equipped with label on outer wall of the injection needle at 60 μm away from the tip.

Further, it is equipped with label on outer wall of the injection needle at 100 μm away from the tip.

Further, the material of the injection needle is glass.

Further, the injection needle is drawn by glass capillary by drawing needle instrument.

Further, there are one the glass columns for axially penetrating through the entire glass capillary for the capillary glass inside pipe wall.

Further, the height of the injection needle is 45~200 μm.

(3) advantageous effect

The above-mentioned technical proposal of the utility model has the advantages that：

1, the gene injection needle of the utility model, simple in structure, shape and inner space are in coniform, in tip Diameter is 0.5 μm, and the internal diameter at 30 μm away from tip is 10 μm, and the internal diameter at 50 μm away from tip is 20 μm, at 45 μm away from tip The volume at place is 3pL；I.e. the injection needle has carried out reasonable setting to parameters such as its height, internal diameter, volumes, thus micro- in progress When operation, the foreign gene that corresponding amount can be drawn according to the position of injection needle high scale (i.e. apart from the height at tip) carries out Injection, can accurately control injection dosage, can safely and effectively reach injection dosage pL class precisions, realize injection foreign gene Quantification.

2, foreign gene is sucked according to the scale on injection needle, ensure that pL grades of precision not by gene fluid viscosity Influence, can safely and effectively reach injection dosage pL class precisions, thus can avoid human factor introduce injection dosage differ and Lead to the problem that embryonic development is inconsistent after injecting, therefore can reduce and mechanical damage is caused to structure in embryo, greatly reduces The death rate of embryo improves transgenosis effect.

3, the injection needle of the utility model is marked on the outer wall, and has measured the volume of the mark so that When carrying out microinjection operation, can be marked using this as reference, avoid larger error when operation, and then improve transgenosis Efficiency.

Description of the drawings

Fig. 1 is the structural schematic diagram of gene injection needle described in the utility model embodiment 1；

Wherein, 1, injection needle；2, the label being equipped on outer wall of the injection needle at 45 μm away from tip.

Specific implementation mode

The embodiment of the utility model is described in further detail with reference to the accompanying drawings and examples.Following embodiment For illustrating the utility model, but cannot be used for limiting the scope of the utility model.

In following embodiment, glass capillary is purchased from Sutter companies of the U.S., article No. BF100-78-10, the capillary Glass tube is cylindrical, outer diameter 1.00mm, internal diameter 0.78mm, length 10cm.There are one axis for the capillary glass inside pipe wall To the glass column through entire glass capillary.It is that the program-controlled horizontal microelectrode of Sutter companies of U.S. P-97 types draws needle to draw needle instrument Instrument.It forges needle instrument and is purchased from Narishige companies of Japan, model MF-900.

Embodiment 1

As shown in Figure 1, present embodiments providing a kind of novel gene injection needle 1, the shape of injection needle 1 and inside Space is in coniform, and the tip internal diameter of injection needle 1 is 0.5 μm, and internal diameter of the injection needle 1 at 30 μm away from tip is 10 μm, note It is 20 μm to penetrate internal diameter of the needle 1 at 50 μm away from tip, and volume of the injection needle 1 at 45 μm away from tip is 3pL.Injection needle 1 away from Outer wall at 45 μm of tip is equipped with label 2.The material of injection needle 1 is glass.The height of injection needle 1 is 70 μm.

Embodiment 2

Present embodiments provide a kind of method preparing gene injection needle described in embodiment 1 comprising following steps：

Step 1：Gene injection needle is drawn using needle instrument is drawn

Glass capillary is installed to and is drawn on needle instrument, drawing needle is carried out, the parameter of needle instrument is drawn to be set as HEAT=650, PULL =30, VEL.=30, TIME=200.

Step 2：The outer wall of gene injection needle is marked using forging needle instrument

According to volume calculations formula V=1/3 π r²(r is radius to h；H is height, i.e., apart from the height at tip), that is, meter It is 0.79pL to calculate and obtain the volume of 10 μm of internal diameter at 30 μm away from tip；The volume of 20 μm of internal diameter is 5.2pL at 50 μm away from tip；Away from At 45 μm of tip, the volume that internal diameter is 16 μm is 3pL.Mark the scale of injection needle outer wall, eyepiece micro- using forging needle instrument Scale lowers monobloc forging needle instrument glass marble longitudinal direction point of contact at 45 μm of scale；The injection needle is disposed vertically in glass marble side, position At 45 μm of scale.Forging needle instrument parameter is set as HEAT=40.Injection needle outer wall and glass marble are lightly contacted when label, Then heat platinum wire, it is seen that the glass tube of injection needle and glass marble be just pasted together and it is indeformable when, stop heating at once, Injection needle is withdrawn rapidly, is marked in the glass pipe outer wall at 45 μm away from tip of injection needle at this time.

Step 3：The injection dosage corresponding to label formed to step 2 is verified

The culture dish equipped with paraffin oil prepared is placed under microscope, injection needle is drawn into paraffin oil to the label Place, then gets the liquid in injection needle, 18 μm of the diameter of drop is measured with eyepiece micro scale, according to ball volume V=1/ 6·πd³(d is diameter), calculating injection dosage are 3pL.

In conclusion the gene injection needle of the utility model, simple in structure, shape and inner space are in circular cone Shape, tip internal diameter are 0.5 μm, and internal diameter 30 μm away from tip at is 10 μm, and the internal diameter at 50 μm away from tip is 20 μm, away from Volume at 45 μm of tip is 3pL；I.e. the injection needle has carried out reasonable setting to parameters such as its height, outer diameter, volumes, thus exists When carrying out micromanipulation, the external source of corresponding amount can be drawn according to the position of injection needle high scale (i.e. apart from the height at tip) Gene is injected, and injection dosage can be accurately controlled, and can safely and effectively be reached injection dosage pL class precisions, be realized injection The quantification of foreign gene.

Foreign gene is sucked according to the scale on injection needle, ensure that pL grades of precision not by the shadow of gene fluid viscosity It rings, thus can avoid the injection dosage that human factor introduces and differ and lead to the problem that embryonic development is inconsistent after injecting, therefore It can reduce and mechanical damage is caused to structure in embryo, greatly reduce the death rate of embryo, improve transgenosis effect.

Moreover, injection needle is marked on the outer wall, and the volume of the mark is measured so that carrying out micro- note It when penetrating operation, can be marked using this as reference, avoid larger error when operation, and then improve transgene efficiency.It utilizes The injection needle for drawing needle instrument to draw, the more conducively microinjection of the label of scale and embryo.

The embodiments of the present invention provide for the sake of example and description, and are not exhaustively or to incite somebody to action The utility model is limited to disclosed form.Many modifications and variations are aobvious and easy for the ordinary skill in the art See.Selection and description embodiment are and to make this field to more preferably illustrate the principles of the present invention and practical application Those of ordinary skill is it will be appreciated that various embodiments with various modifications of the utility model to design suitable for special-purpose.

Claims (9)

1. a kind of novel gene injection needle, which is characterized in that the shape of the injection needle and inner space are in circular cone The tip internal diameter of shape, the injection needle is 0.5 μm, and internal diameter of the injection needle at 30 μm away from the tip is 10 μm, described Internal diameter of the injection needle at 50 μm away from the tip is 20 μm, and volume of the injection needle at 45 μm away from the tip is 3pL.

2. gene injection needle according to claim 1, which is characterized in that the injection needle is at 45 μm away from the tip The outer wall at place is equipped with label.

3. gene injection needle according to claim 1, which is characterized in that the injection needle is at 50 μm away from the tip The outer wall at place is equipped with label.

4. gene injection needle according to claim 1, which is characterized in that the injection needle is at 60 μm away from the tip The outer wall at place is equipped with label.

5. gene injection needle according to claim 1, which is characterized in that the injection needle is away from 100 μ of the tip Outer wall at m is equipped with label.

6. gene injection needle according to claim 1, which is characterized in that the material of the injection needle is glass.

7. gene injection needle according to claim 1, which is characterized in that the injection needle is passed through by glass capillary Needle instrument is drawn to draw.

8. gene injection needle according to claim 7, which is characterized in that there are one axis for the capillary glass inside pipe wall To the glass column through the entire glass capillary.

9. gene injection needle according to claim 1, which is characterized in that the height of the injection needle is 45~200 μ m。