CN112625907B - Embryo long-time culture system - Google Patents

Embryo long-time culture system Download PDF

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Publication number
CN112625907B
CN112625907B CN202011605277.9A CN202011605277A CN112625907B CN 112625907 B CN112625907 B CN 112625907B CN 202011605277 A CN202011605277 A CN 202011605277A CN 112625907 B CN112625907 B CN 112625907B
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embryo
liquid
culture
groove
sample injection
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CN112625907A (en
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肖冬根
王红梅
曾维俊
于福鑫
肖振宇
孙海旋
杨语谌
梁伟国
杨森
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Institute of Zoology of CAS
Suzhou Institute of Biomedical Engineering and Technology of CAS
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Institute of Zoology of CAS
Suzhou Institute of Biomedical Engineering and Technology of CAS
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Abstract

The invention discloses an embryo long-time culture system, which comprises: at least one nutrient solution bottle, at least one liquid path unit connected with the nutrient solution bottle, and at least one waste liquid bottle connected with the liquid path unit; the liquid path unit comprises a sample injection electromagnetic valve connected with the nutrient solution bottle, a sample injection pump connected with the sample injection electromagnetic valve, a liquid discharge electromagnetic valve connected with the sample injection pump, a culture dish assembly connected with the liquid discharge electromagnetic valve and a liquid drawing pump connected with the culture dish assembly, wherein the output end of the liquid drawing pump is connected with the waste liquid bottle. The invention adopts open environment culture, can supplement oxygen in time, avoid generating bubbles; (2) The invention can perform dynamic culture, provide shearing force for the growth and development of embryo, and better simulate the growth and development environment of embryo in the parent; (3) The invention can realize automatic liquid exchange and liquid supplementing, and avoid the influence of manual liquid exchange and liquid supplementing on embryo growth and development; (4) The invention can realize long-time culture of embryo.

Description

Embryo long-time culture system
Technical Field
The invention relates to the field of embryo in-vitro culture, in particular to an embryo long-term culture system.
Background
In vitro embryo culture refers to the process of extracting fertilized eggs or blastula from the inside of an organism, and under the conditions of no bacterial interference, reasonable growth temperature and sufficient needed nutrition, the life conditions of the embryo in the organism are simulated, so that the embryo can perform life activities such as growth, proliferation and the like, thereby realizing in vitro inoculation.
The existing embryo culture device has the following problems: (1) dynamic culture cannot be realized; (2) Automatic replenishment and replacement of culture fluid cannot be realized in the culture process; (3) insufficient oxygen supply; (4) the bubbles affect embryo development and observation.
Disclosure of Invention
The technical problem to be solved by the invention is to provide an embryo long-term culture system aiming at the defects in the prior art. The invention provides an embryo long-time culture system, which solves the problems that the automatic liquid supplement, liquid exchange, dynamic culture and the like cannot be realized in the conventional embryo culture, can realize the long-time culture of embryos, and can create conditions for in-vitro inoculation.
In order to achieve the above purpose, the invention adopts the following technical scheme: an embryo long-term culture system comprising: at least one nutrient solution bottle, at least one liquid path unit connected with the nutrient solution bottle, and at least one waste solution bottle connected with the liquid path unit;
The liquid path unit comprises a sample injection electromagnetic valve connected with the nutrient solution bottle, a sample injection pump connected with the sample injection electromagnetic valve, a liquid discharge electromagnetic valve connected with the sample injection pump, a culture dish assembly connected with the liquid discharge electromagnetic valve and a liquid extraction pump connected with the culture dish assembly, wherein the output end of the liquid extraction pump is connected with the waste liquid bottle.
Preferably, the culture dish assembly comprises a culture dish, a plurality of embryo hoods uniformly arranged in the culture dish at intervals along a straight line, a T-shaped liquid inlet distribution block communicated with the liquid discharge electromagnetic valve through a pipeline and arranged on the first side of the embryo hoods, a liquid inlet channel arranged in the liquid inlet distribution block, a plurality of liquid outlet channels communicated with the liquid inlet channels, a sample feeding needle connected with the liquid outlet channels and a liquid discharge needle arranged on the second side of the embryo hoods.
Preferably, the sample adding needles are in one-to-one correspondence with the embryo hoods, the outlet ends of the sample adding needles are horizontally arranged and are opposite to the first side of the embryo hoods, and the inlet ends of the liquid draining needles are close to the inner wall of the bottom of the culture dish.
Preferably, the embryo cover is used for restraining the embryo in the cavity in the embryo cover, the embryo cover is of a hollowed-out porous structure, a plurality of open holes are formed in the outer wall of the embryo cover, and the open holes have a size enabling the embryo to pass through.
Preferably, the culture dish assembly further comprises a fixing seat, wherein a through groove is formed in the middle of the fixing seat in a penetrating mode along the vertical direction, and a mounting groove communicated with the through groove is formed in the surface of the first side of the fixing seat; a supporting bar fixedly connected with the inner wall of the through groove is arranged in the through groove, the through groove is divided into a sample adding groove and an observing groove by the supporting bar, and the observing groove is positioned right above the embryo covers;
The T-shaped liquid inlet distribution block comprises a sample injection part and a distribution part vertically connected with the sample injection part, the distribution part is connected with the sample adding needles, the sample injection part is matched and arranged in the mounting groove, the distribution part is lapped on the support bar, and the sample adding needles penetrate through the sample adding groove and extend into the culture dish.
Preferably, two sides of the mounting groove are vertically penetrated and provided with two round holes, a bushing is fixedly arranged in the round holes, and a positioning pin is slidably inserted in the bushing;
the bottom surface of the first side of the fixed seat is provided with an arc-shaped sinking groove, and the two round holes are respectively communicated with two sides of the sinking groove;
The spring wire is arranged in the sinking groove, the spring wire is restrained in the sinking groove through a plurality of pressing blocks which are arranged below the sinking groove and connected with the fixing seat, and the distance from the position closest to the center of the round hole on the spring wire is smaller than the radius of the round hole, so that after the locating pin is inserted into the bushing, the spring wire can be in contact with the locating pin, and an extrusion effect is generated on the locating pin.
Preferably, the fixing seat is further provided with a mounting hole for arranging the liquid discharge needle.
Preferably, the sample adding pump is a plunger pump, and the liquid extracting pump is a peristaltic pump.
Preferably, the nutrient solution bottle further comprises a heating device for heating the culture solution discharged from the nutrient solution bottle.
Preferably, the device further comprises a deionized water bottle and a cleaning electromagnetic valve connected with the deionized water bottle, wherein the cleaning electromagnetic valve is connected with the sample injection electromagnetic valve of the liquid path unit.
The beneficial effects of the invention are as follows:
(1) The embryo long-term culture system adopts open environment culture, can supplement oxygen in time, avoids generating bubbles, and is beneficial to observing embryo growth and development and culture process;
(2) The invention can perform dynamic culture, provide shearing force for the growth and development of embryo, and better simulate the growth and development environment of embryo in the parent;
(3) The invention can realize automatic liquid exchange and liquid supplementing, and avoid the influence of manual liquid exchange and liquid supplementing on embryo growth and development;
(4) The invention can realize long-time culture of embryo.
Drawings
FIG. 1 is a schematic diagram showing the structure of an embryo long-term culture system according to example 1 of the present invention;
FIG. 2 is a schematic diagram showing the structure of a culture dish assembly in example 1 of the present invention;
FIG. 3 is a schematic structural view of an embryo cover according to one form of embodiment 1 of the present invention;
FIG. 4 is a schematic structural view of an embryo cover according to another form of embodiment 1 of the present invention;
FIG. 5 is a schematic diagram showing the structure of an embryo long-term culture system according to example 2 of the present invention;
FIG. 6 is a schematic view showing the structure of a culture dish assembly in example 3 of the present invention;
FIG. 7 is a schematic view of the T-shaped liquid inlet distribution block and the fixing seat in the embodiment 3 of the present invention;
FIG. 8 is a schematic view of the structure of the bottom of the culture dish assembly in example 3 of the present invention (with the culture dish removed);
fig. 9 is a schematic view of the structure of the bottom of the fixing base in embodiment 3 of the present invention;
FIG. 10 is a schematic view showing the mounting of the culture dish assembly in example 3 of the present invention on a culture chamber;
Fig. 11 is a schematic exploded view of the fixing base and the positioning pin in embodiment 3 of the present invention.
Reference numerals illustrate:
1-a nutrient solution bottle; 2-a liquid path unit; 3-a waste liquid bottle; 4, a deionized water bottle; 5, cleaning the electromagnetic valve; 6-a heating device; 7, a liquid inlet valve; 8-a temperature sensor; 9-a humidity sensor;
20-a sample injection electromagnetic valve; 21-a sample adding pump; 22-a liquid discharge electromagnetic valve; 23-a culture dish assembly; 24-liquid pump; 25-a culture chamber;
230-Petri dishes; 231-embryoid cover; 232-T type liquid inlet distribution block; 233-a sample adding needle; 234-a drainage needle; 235-fixing seat; 236-through slot; 237-mounting groove; 238-support bar; 2310—opening holes; 2311-a flange structure; 2320—a sample introduction part; 2321—a distribution section; 2350-round hole; 2351-a bushing; 2352—a locating pin; 2353-sink; 2354-wire spring; 2355-mounting hole; 2356-briquetting; 2360-a sample addition well; 2361-observation groove.
Detailed Description
The present invention is described in further detail below with reference to examples to enable those skilled in the art to practice the same by referring to the description.
It will be understood that terms, such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
Example 1
As shown in fig. 1-2, an embryo long-term culture system of the present embodiment includes: at least one nutrient solution bottle 1, at least one liquid path unit 2 connected with the nutrient solution bottle 1, and at least one waste solution bottle 3 connected with the liquid path unit 2;
The liquid path unit 2 comprises a sample injection electromagnetic valve 20 connected with the nutrient solution bottle 1, a sample injection pump 21 connected with the sample injection electromagnetic valve 20, a liquid discharge electromagnetic valve 22 connected with the sample injection pump 21, a culture dish assembly 23 connected with the liquid discharge electromagnetic valve 22 and a liquid suction pump 24 connected with the culture dish assembly 23, wherein the output end of the liquid suction pump 24 is connected with the waste liquid bottle 3. Wherein each liquid path unit 2 can be independently controlled to work. Under the action of a sample adding pump 21, the culture solution in the nutrient solution bottle 1 can be sequentially input into a culture dish assembly 23 through a sample injection electromagnetic valve 20 and a liquid discharge electromagnetic valve 22; the waste liquid in the culture dish assembly 23 can be pumped and discharged into the waste liquid bottle 3 under the action of the liquid pump 24.
Referring to fig. 1, the embodiment includes a nutrient solution bottle 1, a liquid path unit 2, and a waste solution bottle 3.
Referring to fig. 2, in this embodiment, the culture dish assembly 23 includes a culture dish 230, a plurality of embryo hoods 231 disposed in the culture dish 230 at regular intervals along a straight line, a T-shaped liquid inlet distribution block 232 connected to the liquid discharge solenoid valve 22 via a pipe and disposed on a first side of the embryo hoods 231, one liquid inlet channel disposed in the liquid inlet distribution block, and a plurality of liquid outlet channels (i.e., a 1-to-4 channel in this embodiment, the liquid inlet channels and the liquid outlet channels are not shown), a sample injection needle 233 connected to the liquid outlet channels, and a liquid discharge needle 234 disposed on a second side of the embryo hoods 231.
The sample injection needles 233 are in one-to-one correspondence with the embryo covers 231, that is, a first side of each embryo cover 231 has one sample injection needle 233, and in this embodiment, each embryo cover 231 includes 4 embryo covers 231 arranged in a straight line, and one sample injection needle 233 is configured for each embryo cover 231. The outlet end of the sample injection needle 233 is disposed horizontally and opposite to the first side of the embryo cover 231, so that liquid can enter the embryo cover 231 conveniently, and the inlet end of the liquid discharge needle 234 is close to the bottom inner wall of the culture dish 230. When the liquid is changed, the culture liquid can be divided into multiple paths after entering the T-shaped liquid inlet distribution block 232, the culture dish 230 is injected by the sample injection needle 233, and the redundant culture liquid is discharged by the waste liquid needle, so that the liquid is changed; during dynamic culture, the culture solution enters the T-shaped liquid inlet distribution block 232 and then is divided into a plurality of channels, the liquid inlet is used for carrying out reciprocating perfusion and suction action on the embryo cover 231, the dynamic liquid flow is presented in the embryo cover 231, and the fluid shearing effect is generated on the embryo, so that the dynamic culture function can be realized.
The embryo cover 231 is used for restraining the embryo in the cavity inside the embryo cover 231, the embryo cover 231 is of a hollow porous structure and is of a cylindrical shape, a plurality of openings 2310 are formed in the outer wall of the embryo cover 231, the openings 2310 are of sizes enabling the embryo to pass through, the liquid can freely pass through the embryo cover 231, the embryo is restrained in the liquid all the time, and therefore the embryo can be prevented from exceeding an observation visual field along with the flowing of the liquid when the liquid is replaced. Embryos are cultured in culture dish 230 and held in place by embryo cover 231 for ease of monitoring and observation. The openings 2310 on the outer wall of the embryo cover 231 may be obtained by integrally forming a plurality of openings on the outer wall of the embryo cover 231 (as shown in fig. 3), or integrally forming a hollow frame (as shown in fig. 4), and then winding a layer of microporous net on the hollow frame. In one embodiment, the aperture 2310 of the embryo cage 231 is about 80-100 μm in size, the early embryo is approximately a sphere of 150-200 μm, the embryo cannot pass through the gate hole, and the culture medium inside and outside the embryo cage 231 can be freely exchanged.
In a preferred embodiment, the bottom of the embryo cover 231 has a flange structure 2311 to facilitate connection with the bottom surface of the culture dish 230.
In a preferred embodiment, the loading pump 21 is a plunger pump and the liquid pump 24 is a peristaltic pump. The plunger pump can quantitatively suck the liquid into the plunger, and can quantitatively discharge the liquid in the plunger, and the dynamic culture is realized by the reciprocating liquid suction and liquid discharge of the plunger pump and the peristaltic pump from the culture dish 230.
In a preferred embodiment, the embryo long-term culture system further comprises heating means 6 for heating the culture liquid discharged from the nutrient solution bottle 1. The culture medium may be heated to a suitable temperature for embryo culture by heating means 6.
In a preferred embodiment, the embryo long-term culture system further comprises a deionized water bottle 4 and a cleaning electromagnetic valve 5 connected with the deionized water bottle 4, wherein the cleaning electromagnetic valve 5 is connected with a sample injection electromagnetic valve 20 of the liquid path unit 2. The deionized water bottle 4 and the cleaning electromagnetic valve 5 are used for cleaning a liquid path.
Example 2
Referring to fig. 5, the embodiment includes 6 sets of liquid path units 2, namely, 6-channel embryo culture, including 1 deionized water bottle 4, 5 nutrient solution bottles 1 and one waste liquid bottle 3. The deionized water bottles 4 are connected with the sample injection electromagnetic valves 20 of each group of liquid path units 2, the 5 nutrient solution bottles 1 are also connected with the sample injection electromagnetic valves 20 of each group of liquid path units 2, and the waste liquid bottles 3 are connected with the liquid extraction pumps 24 of each group of liquid path units 2; namely, the 6 groups of liquid path units 2 share 1 deionized water bottle 4, all nutrient solution bottles 1 and one waste liquid bottle 3. In this embodiment, one liquid inlet valve 7 is also connected to each of the 5 nutrient solution bottles 1.
In an alternative embodiment, the temperature sensor 8 and the humidity sensor 9 are also arranged on the culture dish assembly 23 of each set of liquid path units 2, so that the temperature and humidity in the culture dish 230 can be detected.
Based on the above structural design, the embryo long-term culture system of the invention can realize automatic liquid exchange and liquid supplement, static culture and dynamic culture, and the working process is described in detail below by taking example 2 as an example.
1. The cleaning process comprises the following steps:
The first step: in the state of liquid suction and empty dish, a cleaning electromagnetic valve 5 connected with a deionized water bottle 4 and a sample injection electromagnetic valve 20 connected with the cleaning electromagnetic valve 5 are opened, after 40ms, a plunger pump pulses to control a plunger to shorten, deionized water is quantitatively sucked into the plunger pump, and after the pump is stopped for 40ms, the cleaning electromagnetic valve 5 and the sample injection electromagnetic valve 20 are closed;
And a second step of: discharging liquid, namely opening a liquid discharge electromagnetic valve 22 for 40ms, pulsing a plunger pump to control the plunger to extend, quantitatively discharging deionized water (the same as the liquid suction amount) into a culture dish 230, and closing the electromagnetic valve after the pump is stopped for 40 ms;
and a third step of: pumping liquid, namely opening a peristaltic pump 24 to pump the liquid waste for a set time, closing the peristaltic pump, and discharging all the liquid in the culture dish into a liquid waste bottle 3;
fourth step: the first to third steps are repeated several times.
2. The liquid exchange process comprises liquid pumping, liquid suction and liquid discharge;
the first step: pumping liquid, starting any peristaltic pump to pump waste liquid for a set time, and closing the peristaltic pump to drain the waste liquid into a waste liquid bottle 3;
And a second step of: liquid suction is carried out, a liquid inlet valve 7 and a sample injection electromagnetic valve 20 are opened, a plunger pump is controlled to pulse, a plunger is controlled to shorten the quantitative suction of culture solution into the plunger pump, and the liquid inlet valve 7 and the sample injection electromagnetic valve 20 are closed;
And a third step of: draining, opening the draining solenoid valve 22, controlling the plunger pump to pulse, controlling the plunger to extend, quantitatively draining the culture solution (same as the liquid suction amount setting) into the culture dish 230, and then closing the draining solenoid valve 22.
3. Static culture process
All valves and pumps are not operated, the embryo is still cultured, and the growth state of the embryo can be observed through a microscope.
4. Dynamic culture process
In the culturing process, the plunger pump is intermittently controlled to pump liquid and circulate liquid from the culture dish 230, and a circulating and reciprocating acting force is generated on the embryo, so that the effect of dynamic culture is achieved.
The process is as follows:
(1) Opening the discharge solenoid valve 22, controlling the pulse generation of the plunger pump, controlling the shortening of the plunger, and quantitatively sucking the culture solution from the culture dish 230 into the plunger pump at a set speed;
(2) After a certain time interval, controlling the plunger pump to pulse to control the plunger to extend, and quantitatively discharging the culture solution from the plunger (the same as the liquid suction amount) to the culture dish 230 at a set speed;
the process (1) and the process (2) are repeated several times, and then the drain solenoid valve 22 is closed.
Example 3
Referring to fig. 6-11, as a further improvement on the basis of embodiment 1 or embodiment 2, in this embodiment, the culture dish assembly 23 further includes a fixing base 235, a through groove 236 is formed in the middle of the fixing base 235 along a vertical direction, and a mounting groove 237 communicated with the through groove 236 is formed on a surface of a first side of the fixing base 235; a supporting bar 238 fixedly connected with the inner wall of the through groove 236 is arranged in the through groove 236, the through groove 236 is divided into a sample adding groove 2360 and an observing groove 2361 by the supporting bar 238, and the observing groove 2361 is positioned right above the embryo covers 231; so that the embryo in the embryo cover 231 can be observed through the observation groove 2361.
The T-shaped liquid inlet distribution block 232 comprises a sample injection portion 2320 and a distribution portion 2321 vertically connected with the sample injection portion 2320, wherein a plurality of sample injection needles 233 are connected to the distribution portion 2321, the sample injection portion 2320 is cooperatively arranged in the mounting groove 237, the distribution portion 2321 is lapped on the support bar 238, and the plurality of sample injection needles 233 penetrate through the sample injection groove 2360 and extend into the culture dish 230, so that the culture liquid is conveyed.
Wherein, two sides of the mounting groove 237 are vertically penetrated and provided with two round holes 2350, a bushing 2351 is fixedly arranged in the round holes 2350, and a positioning pin 2352 is slidably inserted in the bushing 2351;
The bottom surface of the first side of the fixed seat 235 is provided with an arc-shaped sinking groove 2353, and two round holes 2350 are respectively communicated with two sides of the sinking groove 2353;
The spring wire 2354 is arranged in the sinking groove 2353, the spring wire 2354 is restrained in the sinking groove 2353 through a plurality of pressing blocks 2356 which are arranged below the sinking groove 2353 and connected with the fixing seat 235, the spring wire 2354 is in a pre-tightening state in the sinking groove 2353, and the distance between the position, closest to the center of the round hole 2350, of the spring wire 2354 and the center of the round hole 2350 is smaller than the radius of the round hole 2350, so that after the locating pin 2352 is inserted into the bushing 2351, the spring wire 2354 is contacted with the locating pin 2352, and the locating pin 2352 is extruded.
Wherein, the fixing base 235 is also provided with a mounting hole 2355 for arranging the liquid discharge needle 234.
The whole culture dish assembly 23 needs to be connected to the mounting surface, for example, in one embodiment, the culture dish assembly 23 is integrally installed on the culture chamber 25, referring to fig. 10-11, the lower end of the positioning pin 2352 is connected to the culture chamber 25, the fixing seat 235 is sleeved on the positioning pin 2352 through the bushing 2351, the fixing seat 235 can slide up and down on the positioning pin 2352, the spring wire 2354 can squeeze the outer circular surface of the positioning pin 2352, the gap between the bushing 2351 and the positioning pin 2352 is eliminated, a certain contact stiffness is ensured, and a certain squeezing friction force is generated, so that the fixing seat 235 can stop at any position of the vertical sliding stroke of the positioning pin 2352 without freely falling, and the installation is more reliable.
Although embodiments of the present invention have been disclosed above, it is not limited to the use of the description and embodiments, it is well suited to various fields of use for the invention, and further modifications may be readily apparent to those skilled in the art, and accordingly, the invention is not limited to the particular details without departing from the general concepts defined in the claims and the equivalents thereof.

Claims (6)

1. An embryo long-term culture system, comprising: at least one nutrient solution bottle, at least one liquid path unit connected with the nutrient solution bottle, and at least one waste solution bottle connected with the liquid path unit;
The liquid path unit comprises a sample injection electromagnetic valve connected with the nutrient solution bottle, a sample injection pump connected with the sample injection electromagnetic valve, a liquid discharge electromagnetic valve connected with the sample injection pump, a culture dish assembly connected with the liquid discharge electromagnetic valve and a liquid extraction pump connected with the culture dish assembly, and the output end of the liquid extraction pump is connected with the waste liquid bottle;
The culture dish assembly comprises a culture dish, a plurality of embryo hoods uniformly arranged in the culture dish at intervals along a straight line, a T-shaped liquid inlet distribution block communicated with the liquid discharge electromagnetic valve through a pipeline and arranged on the first side of the embryo hoods, a liquid inlet channel arranged in the liquid inlet distribution block, a plurality of liquid outlet channels communicated with the liquid inlet channels, a sample adding needle connected with the liquid outlet channels and a liquid discharge needle arranged on the second side of the embryo hoods;
The sampling needles are in one-to-one correspondence with the embryo hoods, the outlet ends of the sampling needles are horizontally arranged and are opposite to the first side of the embryo hoods, and the inlet ends of the liquid discharge needles are close to the inner wall of the bottom of the culture dish;
The embryo cover is used for restraining the embryo in a cavity in the embryo cover, the embryo cover is of a hollowed-out porous structure, a plurality of open holes are formed in the outer wall of the embryo cover, and the open holes have a size which enables the embryo to pass through;
when in dynamic culture, the culture solution enters the T-shaped liquid inlet distribution block and then is divided into a plurality of paths, the reciprocating perfusion and the suck-back actions are implemented by the sample adding aiming at the embryo cover, the dynamic liquid flow is presented in the embryo cover, and the fluid shearing effect is generated on the embryo, so that the dynamic culture function is realized;
The culture dish assembly further comprises a fixing seat, a through groove is formed in the middle of the fixing seat in a penetrating mode along the vertical direction, and a mounting groove communicated with the through groove is formed in the surface of the first side of the fixing seat; a supporting bar fixedly connected with the inner wall of the through groove is arranged in the through groove, the through groove is divided into a sample adding groove and an observing groove by the supporting bar, and the observing groove is positioned right above the embryo covers;
The T-shaped liquid inlet distribution block comprises a sample injection part and a distribution part vertically connected with the sample injection part, the distribution part is connected with the sample adding needles, the sample injection part is matched and arranged in the mounting groove, the distribution part is lapped on the support bar, and the sample adding needles penetrate through the sample adding groove and extend into the culture dish.
2. The embryo long-term culture system according to claim 1, wherein two sides of the mounting groove are vertically penetrated and provided with two round holes, a bushing is fixedly arranged in the round holes, and a positioning pin is slidably inserted in the bushing;
the bottom surface of the first side of the fixed seat is provided with an arc-shaped sinking groove, and the two round holes are respectively communicated with two sides of the sinking groove;
The spring wire is arranged in the sinking groove, the spring wire is restrained in the sinking groove through a plurality of pressing blocks which are arranged below the sinking groove and connected with the fixing seat, and the distance from the position closest to the center of the round hole on the spring wire is smaller than the radius of the round hole, so that after the locating pin is inserted into the bushing, the spring wire can be in contact with the locating pin, and an extrusion effect is generated on the locating pin.
3. The embryo long-term culture system according to claim 1, wherein the fixing base is further provided with a mounting hole for setting the drain needle.
4. The embryo long term culture system of claim 1, wherein the loading pump is a plunger pump and the liquid pump is a peristaltic pump.
5. The embryo long term culture system of claim 1, further comprising heating means for heating the culture fluid discharged from the nutrient fluid bottle.
6. The embryo long-term culture system of claim 1, further comprising a deionized water bottle and a purge solenoid valve connected to the deionized water bottle, the purge solenoid valve being connected to a sample injection solenoid valve of the fluid path unit.
CN202011605277.9A 2020-12-29 2020-12-29 Embryo long-time culture system Active CN112625907B (en)

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