Vitrified refrigeration carrier and use method thereof

A vitrification and carrier technology, applied in biochemical equipment and methods, biochemical instruments, biological material sampling methods, etc., can solve problems such as inability to isolate liquid nitrogen, contamination of liquid nitrogen and samples by liquid nitrogen, and cross-contamination. Achieve the effect of reducing liquid nitrogen pollution, rapid operation, and improving freezing effect

Active Publication Date: 2010-03-17
陈子江
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0002] At present, there are many kinds of vitrification containers used to preserve biological substances, such as Cryoloop, Cryoleaf, Cryotip, electron microscope grid, etc. Their application effects are similar, but they all have a common disadvantage, that is, they cannot isolate liquid nitrogen
After the cells are loaded into the carrier, the cells are directly in contact with liquid nitrogen, which is likely to cause liquid nitrogen contamination and cross-contamination between liquid nitrogen and samples
At present, those skilled in the art have made many attempts to vitrification carriers, such as using elongated vessels or glass capillaries as carriers for cells or embryos. Although they have achieved certain success, they still cannot completely solve the problem of freezing rate and Liquid nitrogen pollution and other issues

Method used

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  • Vitrified refrigeration carrier and use method thereof
  • Vitrified refrigeration carrier and use method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1: Freezing of human oocytes in vitrified carriers

[0022] (1) Put 2-3 pieces of human oocytes in the composition of modified human fallopian tube fluid (mHTF) + 20% serum substitute (sss) + 7.5% ethylene glycol (EG) + 7.5% dimethyl sulfoxide (DMSO) ) liquid for 4-5 minutes, transferred to the final freezing liquid (mHTF+20% sss+15% EG+15% DMSO+0.5M sucrose (Sucrose);

[0023] (2) Inhale the oocyte together with a small amount of freezing liquid into a Pasteur dropper with an outer diameter of 180±20 μm and an inner diameter of 160±20 μm;

[0024] (3) Insert the head of the thinned Pasteur dropper into the closed inner ring 4 along the internal diameter of the microtube 3 of 220 μm, and place the oocyte and a small amount of freezing liquid on the polyethylene film 2 in the ring;

[0025] (4) Partially immerse the inner ring 4 with the oocyte, the inner microtube 3 and the inner cap 1 in the liquid nitrogen in the liquid nitrogen container, wherein the upper p...

Embodiment 2

[0026] Example 2: Vitrification of human 8-cell embryos in the semi-open arcuate inner ring

[0027] (1) Treat 2-3 human 8-cell embryos in a liquid consisting of mHTF+20% sss+7.5% EG+7.5% PROH (1,2 propylene glycol) for 4-5 minutes, then transfer to the final freezing liquid (mHTF+20%sss+15%EG+15%PROH+0.5M Sucrose);

[0028] (2) Inhale the embryo together with a small amount of freezing liquid into a Pasteur dropper with an outer diameter of 180±20 μm and an inner diameter of 160±20 μm;

[0029] (3) Insert the head of the thinned Pasteur dropper into the semi-open arched inner ring 4 along the inner microtube 3 with an inner diameter of 220 μm, and place the embryo and a small amount of freezing liquid on the polypropylene film 2 in the ring ;

[0030] (4) The inner ring 4 with the embryo, the inner microtube 3 and the inner cap 1 are partly immersed in the liquid nitrogen in the liquid nitrogen container, and then the outer tube 7 is also immersed in the liquid nitrogen. T...

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Abstract

The invention relates to a vitrified refrigeration carrier and a use method thereof for preserving biological substances. The vitrified refrigeration carrier comprises an inner micropipe (3), an innercap (1), an inner ring (4), a film (2), an outer tube (7), an outer tube hole (6) and an outer cap (5), wherein the upper opening of the inner micropipe (3) is connected with the inner cap (1) and passes through the inner cap (1) to be communicated with the outside and closed by the outer cap (5), and the lower opening of the inner micropipe (3) is connected with the inner ring (4). The carrier avoids the direct contact between biological substances and liquid nitrogen, reduces liquid nitrogen pollution and has the advantage of rapid, safe and convenient operation.

Description

technical field [0001] The invention relates to a freezing carrier for storing biological substances and a method for using the same, in particular to a vitrification carrier for storing biological substances and a method for using the same. Background technique [0002] At present, there are many kinds of vitrification containers used to preserve biological substances, such as Cryoloop, Cryoleaf, Cryotip, electron microscope grid, etc. Their application effects are similar, but they all have a common disadvantage, that is, they cannot isolate liquid nitrogen. After the cells are loaded into the carrier, the cells are directly in contact with the liquid nitrogen, which is likely to cause liquid nitrogen contamination and cross-contamination between the liquid nitrogen and the sample. At present, those skilled in the art have made many attempts to vitrification carriers, such as using elongated vessels or glass capillaries as carriers for cells or embryos. Although they have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/24C12N5/00
CPCC12M23/38C12M33/02
Inventor 陈子江吴克良李媛
Owner 陈子江
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