CN218710493U - Microinjection device for gene knock-in - Google Patents

Abstract

The utility model discloses a microinjection device for knocking-in genes, which aims to provide a microinjection device for knocking-in genes, which can push cells to flow towards the impact of a microinjection needle. It includes the injection case, install input tube and injection body on the injection case, be equipped with the miniflow way one of intercommunication in the injection case, miniflow way two, miniflow way three and injection chamber, install in the miniflow way two and take out the inserted bar, take out and install the extrusion piece on the inserted bar, install the movable block in the injection chamber, the syringe needle one end of installation on the injection body is passed the injection chamber and can be dismantled with the movable block and be connected the other end and arrange in outside the injection body, the syringe needle can be dismantled with the injection body and be connected, input tube and miniflow way one intercommunication. The beneficial effects of the utility model are that: the purpose of pushing the cells to flow towards the micro-injection needle only in an impact way can be achieved; the structure is provided with a structure which leads the cell culture solution to be impacted and flowed; the effect of starting the movement of the mobile block is achieved; the injection needle can be conveniently inserted into the moving block; the limiting work of the injection needle can be carried out.

Description

Microinjection device for gene knock-in

Technical Field

The utility model relates to the technical field of biological research, in particular to a microinjection device for knocking in genes.

Background

In particular, the present invention relates to a method and apparatus for biological research, which is a method for manipulating cells or early embryos by using a micromanipulator, a mechanical device for controlling the movement of a micro-needle within the field of a microscope under a high power inverted microscope, so that the gene of a living being can be injected into the cells or early embryos by the micro-injection device, thereby inheriting the gene, and optionally continuing the superior gene.

Chinese patent grant publication no: CN105420099B, entitled bulletin date 2018, 05 and 25, discloses a high-efficiency microinjection device, which comprises a quartz substrate and an injection body, wherein the injection body is made of polydimethylsiloxane material, the injection body is fixed on the quartz substrate, the injection body is provided with an input microchannel, an output microchannel and a cell conveying microchannel for communicating the input microchannel and the output microchannel, a microinjection channel and a cell movement control channel are also arranged between the input microchannel and the output microchannel on the injection body, the microinjection channel is in a vertical direction and is vertically communicated with the cell conveying microchannel, a microinjection needle is fixed on the microinjection channel, a gap is arranged between the microinjection channel and the microinjection needle, the gap size is smaller than the cell diameter, a limiting bulge for limiting the cell movement is arranged in the microinjection channel, the cell movement control channel is vertically communicated with the microinjection channel, and the cell movement control channel is positioned above the limiting bulge; the upper part of the injection body is not provided with a fixing plate for fixing the micro-injection needle, and the fixing plate is made of polydimethylsiloxane material. The technical scheme has the defects that in the injection working process, cells are driven by microfluid to move to an inverted funnel-shaped area in the middle of a microinjection flow channel and then are limited, the microinjection needle is penetrated to carry out injection working, but the size of the cells is small and different, so that the limiting work of each cell cannot be smoothly carried out by the funnel area with a fixed diameter, and then under the pushing effect of the microinjection needle, the cells can be influenced by the pushing effect and are separated from the limitation, and the injection working cannot be smoothly carried out.

In summary, the cell can only impact and flow against the area where the microinjection needle is located, so that the cell can directly impact the microinjection needle to inject the cell, and the injection is further smoothly driven by the impulse force.

SUMMERY OF THE UTILITY MODEL

The utility model provides a microinjection device for knocking in gene, which can push cells to flow towards the microinjection needle impact, in order to overcome the defect that the spacing work of each cell can not be smoothly carried out in the prior art.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

the utility model provides a microinjection device for gene is knocked in, includes the injection case, install input tube and injection body on the injection case, be equipped with miniflow way one, miniflow way two, miniflow way three and injection chamber in the injection case, miniflow way one, miniflow way two and injection chamber all communicate with miniflow way three, miniflow way one and miniflow way two are arranged in the left and right both ends of miniflow way three respectively, the injection chamber is arranged in the upper end of miniflow way three, install the extraction and insertion pole in miniflow way two, the extrusion piece is installed to the one end of extraction and insertion pole, the outward flange of extrusion piece and miniflow way two phase contact, the other end of extraction and insertion pole passes miniflow way two and arranges outside the injection case, install the movable block in the injection chamber, the movable block is connected with the injection case activity, be equipped with the syringe needle on the injection body, install the syringe needle on the injection body, one end of syringe needle passes the injection chamber and can dismantle the connection with the movable block and be connected, the other end of syringe needle passes outside the injection body, the input tube is communicated with miniflow way one.

The injection box is provided with a micro-channel I, a micro-channel II, a micro-channel III and an injection cavity, wherein the micro-channel I, the micro-channel II and the injection cavity are communicated with the micro-channel III, the micro-channel I and the micro-channel II are respectively arranged at the left end and the right end of the micro-channel III, the injection cavity is arranged at the upper end of the micro-channel III, the cell culture fluid flows in the micro-channel I, the micro-channel II and the injection cavity, the end part of a draw rod outside the injection box can be controlled timely, the extrusion block in the micro-channel II moves towards the micro-channel III, an impact effect is generated on the cell culture fluid, the two ends of the cell culture fluid can only flow towards the impact effect of the extrusion block under the matching of the injection effect of the injection pipe, the cell culture fluid flows towards the micro-channel III, the injection fluid can be pushed to move towards the upper end of the injection box after flowing to the position of the injection cavity, and the cell culture fluid can only flow towards the upper end of the injection block, and the moving block can be pushed out from the injection cavity.

Preferably, the moving block is provided with a penetrating hole, the position of the penetrating hole corresponds to the position of the injection needle connector up and down, two limiting blocks are installed at the penetrating hole, the two limiting blocks are bilaterally symmetrical by taking the penetrating hole as a center, and the limiting blocks are detachably connected with the moving block. Design like this through the interlude hole on the movable block alright let the movable block when moving towards injection case upper end direction, the smooth insertion of syringe needle on the injection body, the position of this interlude hole corresponds from top to bottom with the position of syringe needle connector just the syringe needle can insert structural design smoothly, two stoppers are installed to interlude hole department, two stoppers use the interlude hole as central bilateral symmetry, and stopper and movable block can dismantle to be connected then when the syringe needle inserts the interlude hole, two stoppers are promoted by the syringe needle and are reversed to move away from, just so form the passageway that only has the syringe needle, cell culture solution just can flow towards the passageway that has the syringe needle, develop the injection work that the syringe needle inserted the cell.

Preferably, the moving block is provided with two moving grooves, the moving grooves correspond to the limiting blocks one to one, one end of each limiting block is arranged outside the corresponding moving groove, the other end of each limiting block is provided with a spring, the springs are arranged in the corresponding moving grooves, and the limiting blocks are connected with the moving block through the springs. The design can install the spring separately through two shifting grooves on the movable block like this, lets spring and stopper do well and be connected, so outside the one end of stopper just can arrange the shifting groove in, the other end then does well through spring and movable block and is connected, two stoppers receive the promotion of syringe needle to reverse move away from just to go on steadily under the elastic effect of spring, and break away from the promotion work of syringe needle, the elastic effect of spring just can let two stoppers remove in opposite directions, until bump together.

Preferably, a switch is installed in the moving groove, the switch is sleeved in the spring, an alarm bell is installed on the injection box, and the alarm bell is electrically connected with the switch. The switch can be installed through the shifting chute to the design like this, and the switch suit can be in the spring, and two stoppers are by the reverse alright touch switch of puting aside of the promotion of syringe needle, let the alarm bell of being connected with the switch electricity make a sound, remind injection work of having begun, in time change the syringe needle after the injection work of being convenient for finishes.

Preferably, the junction of the first micro-channel and the third micro-channel and the junction of the second micro-channel and the third micro-channel are both in an acute angle alpha, the lower end of the third micro-channel is provided with an outlet, the outlet is communicated with the third micro-channel, the outlet is provided with a plug, and the plug is detachably connected with the injection box. Design like this through the junction of miniflow channel one and miniflow channel three and miniflow channel two and the junction of miniflow channel three all is acute angle alpha's structure, after the injection work of being convenient for, the cell of injection is towards the reverse flow of the lower extreme of miniflow channel three under the drive of cell culture liquid, and the lower extreme of miniflow channel three is equipped with the export, the stopper in exit can dismantle with the injection case and be connected, after the injection work so, export is opened to steerable stopper, let cell culture liquid discharge smoothly, be convenient for collect the cell after the injection.

Preferably, be equipped with a plurality of syringe groove on the injection case, the syringe groove includes notch and tank bottom, the tank bottom department installs the grafting piece, the syringe needle contains a plurality of, syringe needle and syringe groove phase-match. The design can insert the syringe needle through a plurality of syringe needle groove one by one on the injection case like this, and the syringe needle groove includes notch and tank bottom, and insert on the plug-in block of tank bottom department can be inserted to the syringe needle, lets the syringe needle receive spacingly, reduces the probability that the syringe needle breaks away from the injection case.

The utility model has the advantages that: the purpose of pushing the cells to flow towards the micro-injection needle only in an impact way can be achieved; the structure is provided with a structure which leads the cell culture solution to be impacted and flowed; the effect of starting the movement of the mobile block is achieved; the injection needle can be conveniently inserted into the moving block; the limiting work of the injection needle can be carried out.

Drawings

Fig. 1 is a schematic structural view of the present invention;

fig. 2 is a schematic view of an operating state of the present invention;

fig. 3 is a schematic view of another working state of the present invention;

fig. 4 is a schematic structural view of a section a in fig. 3.

In the figure: 1. injection case, 2, input tube, 3, export, 4, stopper, 5, miniflow way one, 6, miniflow way two, 7, miniflow way three, 8, injection chamber, 9, movable block, 10, injection body, 11, syringe needle connector, 12, syringe needle, 13, syringe needle groove, 14, grafting piece, 15, extraction and insertion pole, 16, extrusion piece, 17, interlude hole, 18, movable groove, 19, stopper, 20, spring, 21, switch, 22, alarm bell.

Detailed Description

The following description of the present invention will be made with reference to the accompanying drawings.

In the embodiment shown in fig. 1, 2 and 3, a microinjection device for knocking in genes comprises an injection box 1, an input tube 2 and an injection body 10 are installed on the injection box 1, a first microchannel 5, a second microchannel 6, a third microchannel 7 and an injection cavity 8 are arranged in the injection box 1, the first microchannel 5, the second microchannel 6 and the injection cavity 8 are all communicated with the third microchannel 7, the first microchannel 5 and the second microchannel 6 are respectively arranged at the left end and the right end of the third microchannel 7, the injection cavity 8 is arranged at the upper end of the third microchannel 7, a drawing rod 15 is installed in the second microchannel 6, an extrusion block 16 is installed at one end of the drawing rod 15, the outer edge of the extrusion block 16 is in contact with the second microchannel 6, the other end of the drawing rod 15 penetrates through the second microchannel 6 and is arranged outside the injection box 1, a moving block 9 is installed in the injection cavity 8, the moving block 9 is movably connected with the injection box 1, a connecting port 11 is arranged on the injection body 10, an injection needle 12 is installed on the injection body 10, one end of the injection needle 8 penetrates through the injection cavity 8 and is connected with the injection needle 10, and the injection needle 10 and the injection tube 10 is communicated with the injection needle 5.

As shown in fig. 1 and 4, a through-hole 17 is formed in the moving block 9, the through-hole 17 is located at a position corresponding to the position of the injection needle connector 11 up and down, two limiting blocks 19 are installed at the through-hole 17, the two limiting blocks 19 are bilaterally symmetrical about the through-hole 17, and the limiting blocks 19 are detachably connected with the moving block 9. Two moving grooves 18 are formed in the moving block 9, the moving grooves 18 correspond to the limiting blocks 19 one to one, one ends of the limiting blocks 19 are arranged outside the moving grooves 18, springs 20 are installed at the other ends of the limiting blocks 19, the springs 20 are arranged in the moving grooves 18, and the limiting blocks 19 are connected with the moving block 9 through the springs 20. A switch 21 is arranged in the moving groove 18, the switch 21 is sleeved in a spring 20, an alarm bell 22 is arranged on the injection box 1, and the alarm bell 22 is electrically connected with the switch 21.

As shown in fig. 1 and 2, the connection part of the first micro-channel 5 and the third micro-channel 7 and the connection part of the second micro-channel 6 and the third micro-channel 7 are both in an acute angle alpha, the lower end of the third micro-channel 7 is provided with an outlet 3, the outlet 3 is communicated with the third micro-channel 7, a plug 4 is arranged at the outlet 3, and the plug 4 is detachably connected with the injection box 1. Be equipped with a plurality of syringe groove 13 on injection case 1, syringe groove 13 includes notch and tank bottom, and the groove department installs plug 14, and syringe needle 12 contains a plurality of, syringe needle 12 and syringe groove 13 phase-match.

Firstly, an extrusion block 16 on a drawing and inserting rod 15 is placed into a second microchannel 6 in an injection box 1, the outer edge of the extrusion block 16 is in contact with the wall of the second microchannel 6, in order to enhance the sealing performance of the second microchannel 6, then an injection needle 12 is inserted into an injection cavity 8 in the injection box 1 through an injection needle connecting port 11 on an injection body 10, the position of the injection needle connecting port 11 is vertically corresponding to the position of a through hole 17 on a moving block 9 in the injection cavity 8, then a cell culture solution containing cells to be injected can be injected through an input tube 2 arranged on the injection box 1, the cell culture solution is led into a first microchannel 5 communicated with the input tube 2, and in the injection box 1, the first microchannel 5, the second microchannel 6 and the injection cavity 8 are communicated with a third microchannel 7, therefore, the first microchannel 5 and the second microchannel 6 are respectively placed at the left end and the right end of the third microchannel 7, the injection cavity 8 is arranged at the upper end of the micro-channel three 7, the injection of the cell culture fluid in the micro-channel one 5 preferentially flows at the bottom ends of the micro-channel one 5, the micro-channel two 6 and the micro-channel three 7, the end part of the inserting and pulling rod 15 outside the injection box 1 is controlled in the continuous injection process of the cell culture fluid, the extrusion block 16 on the inserting and pulling rod 15 moves towards the direction of the micro-channel three 7, so that the micro-channel two 6 has the impulsive force for pushing the cell culture fluid, the cell culture fluid flows towards the direction of the injection cavity 8 under the matching of the injection impulsive force of the micro-channel one 5 and the extrusion block 16 of the micro-channel two 6 until the moving block 9 in the injection cavity 8 is pushed to move towards the injection body 10, wherein the micro-channel one 5 and the micro-channel two 6 both comprise two flow channels, the diameter of the flow channel communicated with the injection box 1 can be larger than the diameter of the flow channel communicated with the micro-channel three 7, the diameter of the third micro-channel 7 is equal to the diameter of the first micro-channel 5 communicated with the third micro-channel 7, so that the impact force at the first micro-channel 5 and the second micro-channel 6 is enhanced, and the impact force of the cell culture solution on the moving block 9 is stronger.

Under the impact effect of the cell culture solution, the moving block 9 can move towards the injection body 10 in the injection cavity 8, the outer edge of the injection body 10 can be well contacted with the inner wall of the injection cavity 8 to enhance the sealing performance of the injection cavity 8, the upper end of the injection needle 12 is arranged outside the injection body 10, the lower end of the injection needle 12 can be provided with a needle head, in the moving process of the moving block 9, the needle head is firstly inserted into the insertion hole 17 and then is contacted with the two limiting blocks 19 arranged on the moving block 9 to move towards the two limiting blocks 19, so that a channel in which the needle head is inserted is opened, the cell culture solution can only flow from the channel direction, and the purpose of pushing cells to impact and flow only towards the microinjection needle is achieved. The insertion holes 17 may be formed such that the upper ends thereof are flush with the upper end of the movable block 9, and the lower ends of the insertion holes 17 are formed with grooves, so that the cell culture solution can be smoothly collected and flow toward the insertion holes 17 from the grooves. Furthermore, one end of the stopper 19 is disposed outside the moving groove 18, and the other end of the stopper 19 is connected to the moving block 9 through the spring 20, so that the movement of the stopper 19 can be stably performed. In the moving process of the limiting block 19, the switch 21 sleeved by the spring 20 can be touched, and the alarm bell 22 electrically connected with the switch 21 makes a sound to remind that the insertion work is started smoothly.

When the cell culture solution flows into the channel, the cells in the cell culture solution can enter along with the cell culture solution, under the push of the impulse force of the first micro-channel 5 and the impulse force of the second micro-channel 6, the cells can smoothly touch the needle head of the injection needle 12, the needle head is inserted into the cells, and the injection needle 12 outside the injection body 10 can be controlled to inject the injection solution containing the genes, so that the injection work of knocking-in the genes is completed, and the genes which are expected to be injected can be smoothly injected into the cells. And finally, stopping injecting the cell culture solution into the input pipe 2, and controlling the extrusion block 16 to move towards the direction outside the injection box 1, so that the cell culture solution with the cells injected with the genes can flow towards the lower end of the micro-channel III 7, and controlling the plug 4 arranged at the lower end of the micro-channel III 7 to open the outlet 3 at the lower end of the micro-channel III 7 so as to discharge the cell culture solution from the outlet 3, thereby facilitating the collection of the cells injected with the genes. Here, the acute angle α formed between the connection point of the first micro flow channel 5 and the third micro flow channel 7 and the connection point of the second micro flow channel 6 and the third micro flow channel 7 is a structural design that allows the cell culture fluid without the effect of impulse to flow smoothly to the outlet 3. After injection work is finished, the injection needle 12 and the injection body 10 can be detached from each other, so that the two limiting blocks 19 are separated from the pushing limiting effect of the injection needle 12 and move towards each other under the elastic effect of the spring 20 until the injection needle 12 and the injection body are contacted with each other.

For the injection of various genes, the preparation of a plurality of injection needles 12 can be selected, and the spare injection needle 12 is inserted into a matched injection needle groove 13 on the injection box 1 and limited by an insertion block 14 arranged at the bottom of the groove, so that the probability that the injection needle 12 is separated from the injection box 1 can be reduced.

On the microinjection device, the rubber with the roller only arranged at the middle end can be sleeved on the outer edge of the moving block 9 and the outer edge of the extrusion block 16, so that the moving block 9 and the extrusion block 16 can move smoothly and have good sealing performance. The connection between the injection needle 12 and the injection body 10 and the connection between the injection needle 12 and the insertion block 14 can be threaded, which facilitates the stability of the connection and the convenience of detachment.

Claims (6)

1. The utility model provides a microinjection device for gene is knocked in, characterized by, includes injection box (1), install input tube (2) and injection body (10) on injection box (1), be equipped with miniflow way (5), miniflow way two (6), miniflow way three (7) and injection chamber (8) in injection box (1), miniflow way one (5), miniflow way two (6) and injection chamber (8) all communicate with miniflow way three (7), both ends about miniflow way three (7) are arranged respectively in miniflow way one (5) and miniflow way two (6), the upper end of miniflow way three (7) is arranged in injection chamber (8), install in miniflow way two (6) and take out inserted bar (15), take out the one end of inserted bar (15) and install extrusion block (16), the outward flange and the miniflow way two (6) of extrusion block (16) contact, the other end of taking out inserted bar (15) passes miniflow way two (6) and arranges in injection box (1), install in injection chamber (8) movable block (9), injection chamber (10) and injection chamber (10) are equipped with injection needle (11) connection movable block (10) injection needle (10) injection chamber (10), injection needle (10) are equipped with movable block (10) on injection chamber (10) injection needle (10) injection chamber (11) injection needle) the last movable block (10) is equipped with injection needle (10) is equipped with injection chamber (11) the last ) The detachable connection is realized, the other end of the injection needle (12) penetrates through the injection needle connecting port (11) and is arranged outside the injection body (10), the injection needle (12) is detachably connected with the injection body (10), and the input tube (2) is communicated with the micro-channel I (5).

2. The microinjection device for gene knock-in according to claim 1, wherein the moving block (9) is provided with a through hole (17), the through hole (17) is located at a position corresponding to the position of the injection needle connection port (11) up and down, two limiting blocks (19) are installed at the through hole (17), the two limiting blocks (19) are symmetrical left and right with the through hole (17) as a center, and the limiting blocks (19) are detachably connected with the moving block (9).

3. The microinjection device for gene knockin according to claim 2, wherein the moving block (9) is provided with two moving grooves (18), the moving grooves (18) correspond to the limiting blocks (19) one to one, one end of each limiting block (19) is disposed outside the moving groove (18), the other end of each limiting block (19) is provided with a spring (20), the springs (20) are disposed in the moving grooves (18), and the limiting blocks (19) are connected to the moving block (9) through the springs (20).

4. A microinjection apparatus for gene knockin according to claim 3, wherein a switch (21) is installed in the movable groove (18), the switch (21) is installed in a spring (20) in a sleeved manner, an alarm bell (22) is installed on the injection box (1), and the alarm bell (22) is electrically connected to the switch (21).

5. The microinjection device for gene knock-in according to claim 1, wherein the junction of the first micro flow channel (5) and the third micro flow channel (7) and the junction of the second micro flow channel (6) and the third micro flow channel (7) are both at an acute angle α, the lower end of the third micro flow channel (7) is provided with an outlet (3), the outlet (3) is communicated with the third micro flow channel (7), the outlet (3) is provided with a plug (4), and the plug (4) is detachably connected with the injection box (1).

6. The microinjection device for gene knock-in according to claim 1, wherein the injection box (1) is provided with a plurality of needle grooves (13), the needle grooves (13) include a notch and a groove bottom, an insertion block (14) is installed at the groove bottom, the injection needles (12) comprise a plurality of needles, and the injection needles (12) are matched with the needle grooves (13).

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