Integrated carrier device for freezing and unfreezing embryos and use method of integrated carrier device

Abstract

The integrated carrier device comprises a microporous membrane substrate, a frame and a sealing film, a plurality of through holes are formed in the microporous membrane substrate, the frame is arranged on the edge of the microporous membrane substrate, and the sealing film is arranged at the top end of the frame. The device further comprises a base, and an adsorption block matched with the lower surface of the microporous membrane substrate is fixedly arranged on the upper surface of the base. According to the integrated carrier device for embryo freezing and unfreezing, by changing the structure of the freezing carrier, all key steps are completed on the carrier, embryo transfer is not needed, the operation difficulty and complexity are reduced, the operation steps are reduced, time is more easily and accurately controlled, and the probability that due to different proficiency degrees of an operator, the operation time is shortened is reduced. And effect differences caused by random errors of the operation process.

Description

Technical Field

[0001] This invention relates to the field of assisted reproductive technology, and more particularly to an integrated carrier device for embryo freezing and thawing. Background Technology

[0002] Embryo (including oocyte) freezing / thawing is a crucial routine procedure in IVF laboratories. This procedure demands a high level of skill from technicians, especially the final freezing step and the first thawing step, which require precise time control. Current methods involve continuously transferring embryos between different droplets and carriers, with each step performed under a stereomicroscope. This method is complex, challenging, and individual differences significantly impact embryo viability after thawing.

[0003] The traditional method for freezing / thawing embryos involves dripping cryogenic solution into the wells of a four-well (or five-well) plate and continuously transferring the embryos from different liquids and carriers using a pipette. The specific steps are as follows: Freezing process: The embryos are immersed in the first freezing solution (EM) for 5-10 minutes; then transferred to the second freezing solution (VM), and after a short stay, loaded onto the carrier blades; and then rapidly immersed in liquid nitrogen within 60 seconds.

[0004] Thawing process: Remove the carrier from liquid nitrogen and quickly insert it into the first thawing solution (TS) at 37°C; transfer the embryo to the second thawing solution (DS) within 60 seconds, and transfer it to the third thawing solution (WS1 / 2) after 3 minutes. After 5 minutes, transfer it to the regular culture medium and place it in an incubator for later use.

[0005] Existing technologies have the following drawbacks: They are cumbersome, requiring multiple transfers of embryos between droplets and carriers, and are highly dependent on operator skill; timing is difficult to control, as different operators significantly impact embryo viability after cryopreservation and thawing, and fluctuations in the 60-second window period affect embryo survival rates; and they are costly, consuming 500-1000 μL of cryopreservation solution per step. Due to their complexity, automation and intelligentization are difficult to achieve.

[0006] Therefore, how to develop an integrated carrier device for embryo freezing and thawing has become an urgent problem for those skilled in the art. Summary of the Invention

[0007] The purpose of this invention is to provide an integrated carrier device for embryo freezing and thawing, which solves the problem in the prior art that embryos need to be transferred between droplets and carriers multiple times during the embryo freezing or thawing process, and the embryo survival rate depends on the operator's skill.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: The present invention discloses an integrated carrier device for embryo freezing and thawing, comprising a microporous membrane substrate, a frame, and a sealing film. The microporous membrane substrate has several through holes, the edge of the microporous membrane substrate is provided with a frame, and the top of the frame is provided with a sealing film.

[0009] Furthermore, it also includes a base, on the upper surface of which an adsorption block is fixedly disposed to cooperate with the lower surface of the microporous membrane substrate.

[0010] Furthermore, the microporous membrane substrate is elliptical in shape, and the shape and size of the adsorption block are the same as those of the microporous membrane substrate.

[0011] Furthermore, clamping pieces are provided on both sides of the frame.

[0012] Furthermore, the upper surface of the clamping piece is a smooth surface, and the lower surface of the clamping piece is a frosted surface.

[0013] Furthermore, the lower surface of the sealing film is provided with reusable adhesive that mates with the frame and the clamping piece.

[0014] Furthermore, the absorbent block is specifically made of absorbent cotton.

[0015] Furthermore, a label is printed on the sealing film; the sealing film has an embedded RFID chip.

[0016] Furthermore, the base used for the freezing process is provided with three adsorption blocks; the base used for the thawing process is provided with one adsorption block.

[0017] A method of using an integrated carrier device for embryo freezing and thawing includes the following steps: Freezing process: The first step is to transfer the embryo from the culture dish to the microporous membrane substrate of the carrier; The second step is to place the carrier on the first adsorption block of the base used for the freezing process to remove the culture medium. The third step is to cover the embryos with the first freezing solution and let them stand for 5 to 10 minutes. Fourth step: Transfer the carrier to the second adsorption block to absorb the liquid, add the second cryogenic liquid and stir gently; Fifth step: close the sealing membrane, move the carrier to the third adsorption block after 50 seconds, and immerse it in liquid nitrogen after 60 seconds; Thawing process: The first step is to remove the carrier from the liquid nitrogen, place it on a 37°C hot plate, and then remove the sealing film. The second step is to immediately add the first thawing solution at 37°C; Third, after 50 seconds, move the carrier to the adsorption block of the base used for the defrosting process to absorb the liquid. Fourth, add the second thawing solution, and after 3 minutes, transfer the embryos to a culture dish prepared with the third thawing solution, and complete the thawing process according to the usual steps.

[0018] Compared with the prior art, the beneficial technical effects of the present invention are as follows: In this invention, the integrated carrier device for embryo freezing and thawing utilizes a microporous membrane carrier where all critical embryo processing steps are completed on the carrier, eliminating the need for transfer operations. This reduces the amount of critical freezing reagent used from 2000 μL to 300 μL, significantly lowering costs. The absorbent cotton step solidifies the operational rhythm, allowing for more precise time control and reducing operator variability. The closed-loop freezing system also minimizes the risk of potential contamination. In summary, this integrated carrier device for embryo freezing and thawing, by modifying the structure of the freezing carrier, completes all critical steps on the carrier itself, eliminating the need for embryo transfer. This reduces operational difficulty and complexity, streamlines the process, facilitates precise time control, and minimizes differences in results due to operator skill levels and random errors. Attached Figure Description

[0019] The present invention will be further described below with reference to the accompanying drawings: Figure 1 This is an isometric view (excluding the sealing film and base) of the integrated carrier device for embryo freezing and thawing according to the present invention. Figure 2 This is a schematic diagram of a microporous membrane substrate structure; Figure 3 This is a schematic diagram of the sealing film structure; Figure 4 A base for use in thawing operations; Figure 5 A base for use in freezing operations.

[0020] Explanation of reference numerals in the attached drawings: 1. Microporous membrane substrate; 2. Frame; 201. Clamping piece; 3. Sealing film; 4. Base; 401. Adsorption block. Detailed Implementation

[0021] like Figures 1 to 5 As shown, an integrated carrier device for embryo freezing and thawing includes a microporous membrane substrate 1, a frame 2, a sealing membrane 3, and a base 4.

[0022] The microporous membrane substrate 1 has several through holes with a pore size of 10 to 20 μm to match the diameter of an embryo, and is made of a biocompatible polymer (such as COP). The microporous membrane substrate 1 is elliptical in shape.

[0023] The microporous membrane substrate 1 has a frame 2 along its edge. The frame 2 is made of embryo-grade PVP material, and its upper surface protrudes 2 mm above the microporous membrane to accommodate liquid. The lower surface of the frame 2 extends 0.3 mm beyond the microporous membrane. This design prevents the microporous membrane substrate 1 from contacting the work surface, thus avoiding the risk of infection.

[0024] Clamping pieces 201 are provided on both sides of the frame 2. The upper surface of the clamping piece 201 is smooth to increase the adhesion area with the sealing film 3. The lower surface of the clamping piece 201 is frosted to facilitate gripping with tweezers.

[0025] A sealing film 3 is provided at the top of the frame 2. The lower surface of the sealing film 3 is provided with a reusable adhesive that mates with the frame 2 and the clamping piece 201.

[0026] Specifically, the sealing film 3 is a puncture-resistant plastic film with its edges coated with medical-grade reusable adhesive.

[0027] A label is printed on the sealing film 3; the sealing film 3 has a built-in RFID chip.

[0028] It also includes a base 4, on the upper surface of which an adsorption block 401 is fixedly disposed, which mates with the lower surface of the microporous membrane substrate 1. The shape and size of the adsorption block 401 are the same as those of the microporous membrane substrate 1. The adsorption block 401 is specifically made of absorbent cotton.

[0029] In order to facilitate smooth integration between the adsorption block 401 and the microporous membrane substrate 1, the vertical cross-section of the adsorption block 401 is a trapezoid with a smaller top and a larger bottom.

[0030] Three adsorption blocks 401 are provided on the base 4 for the freezing process. Their area and shape are completely in contact with the bottom surface of the carrier, and they are used for liquid adsorption during the freezing process.

[0031] An adsorption block 401 is provided on the base 4 for the thawing process. Its area and shape are completely in contact with the bottom surface of the carrier for liquid adsorption during the thawing process.

[0032] A method of using an integrated carrier device for embryo freezing and thawing includes the following steps: Freezing process: The first step is to transfer the embryo from the culture dish to the microporous membrane substrate 1 of the carrier; The second step is to place the carrier on the first adsorption block 401 of the base 4 used for the freezing process to remove the culture medium. Third, add 100μL of the first freezing solution to cover the embryos and let them stand for 5 to 10 minutes; Fourth step: Transfer the carrier to the second adsorption block 401 to absorb the liquid, add 50 μL of the second cryogenic liquid and stir gently; Step 5: Close the sealing membrane 3. After 50 seconds, move the carrier to the third adsorption block 401. After 60 seconds, immerse it in liquid nitrogen. Thawing process: First, remove the carrier from the liquid nitrogen, place it on a 37°C hot plate, and remove the sealing film 3; The second step is to immediately add 100 μL of the first thawing solution at 37°C; Third step: After 50 seconds, move the carrier to the adsorption block 401 of the base 4 used for the defrosting process to absorb the liquid. Fourth, add the second thawing solution, and after 3 minutes, transfer the embryos to a culture dish prepared with the third thawing solution, and complete the thawing process according to the usual steps.

[0033] In this invention, all key embryo processing steps are completed on a microporous membrane carrier, eliminating the need for transfer operations and reducing the number of transfers from six to zero compared to the traditional method. The volume of crucial freezing reagent is reduced from 2000 μL to 300 μL, significantly lowering costs. The absorbent cotton process solidifies the operational rhythm, allowing for more precise time control and reducing operator variability. This invention employs a closed-loop freezing system, minimizing the risk of potential contamination.

[0034] By simplifying the steps, it is easier to automate freezing / thawing. This invention is an improvement on the preliminary operation steps of the next freezing / solution process.

[0035] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. An integrated carrier device for embryo freezing and thawing, characterized in that: It includes a microporous membrane substrate (1), a frame (2) and a sealing film (3). The microporous membrane substrate (1) has several through holes, the edge of the microporous membrane substrate (1) is provided with a frame (2), and the top of the frame (2) is provided with a sealing film (3).

2. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: It also includes a base (4), on the upper surface of which an adsorption block (401) is fixedly disposed, which cooperates with the lower surface of the microporous membrane substrate (1).

3. The integrated carrier device for embryo freezing and thawing according to claim 2, characterized in that: The microporous membrane substrate (1) is elliptical in shape, and the adsorption block (401) has the same shape and size as the microporous membrane substrate (1).

4. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: Clamping pieces (201) are provided on both sides of the frame (2).

5. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: The upper surface of the clamping piece (201) is a smooth surface, and the lower surface of the clamping piece (201) is a frosted surface.

6. The integrated carrier device for embryo freezing and thawing according to claim 4, characterized in that: The lower surface of the sealing film (3) is provided with a reusable adhesive that mates with the frame (2) and the clamping piece (201).

7. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: The adsorption block (401) is specifically made of absorbent cotton.

8. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: A label is printed on the sealing film (3); the sealing film (3) has an RFID chip built in.

9. The integrated carrier device for embryo freezing and thawing according to claim 1, characterized in that: Three adsorption blocks (401) are provided on the base (4) for the freezing process; one adsorption block (401) is provided on the base (4) for the thawing process.

10. A method of using an integrated carrier device for embryo freezing and thawing, comprising the integrated carrier device for embryo freezing and thawing as described in any one of claims 1 to 9, characterized in that... Includes the following steps, Freezing process: The first step is to transfer the embryo from the culture dish to the microporous membrane substrate (1) of the carrier; The second step is to place the carrier on the first adsorption block (401) of the base (4) used for the freezing process to remove the culture medium; The third step is to cover the embryos with the first freezing solution and let them stand for 5 to 10 minutes. Fourth step: Transfer the carrier to the second adsorption block (401) to absorb the liquid, add the second cryogenic liquid and stir gently; Step 5: Close the sealing membrane (3), and after 50 seconds, move the carrier to the third adsorption block (401), and after 60 seconds, put it into liquid nitrogen; Thawing process: First step, remove the carrier from liquid nitrogen, place it on a 37°C hot plate, and peel off the sealing film (3). The second step is to immediately add the first thawing solution at 37°C; Third step, after 50 seconds, the carrier is moved to the adsorption block (401) of the base (4) used for the defrosting process to absorb the liquid; Fourth step: Add the second thawing solution, and after 3 minutes, transfer the embryos to a culture dish prepared with the third thawing solution, and complete the thawing process according to the standard procedure.

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