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Vitrification freezing observing device and application thereof

A vitrification and observation device technology, applied in the direction of measuring devices, material analysis through optical means, instruments, etc., can solve problems such as inflexibility, ineffective work, and inability to find glass liquid compatibility in time, and achieve the effect of reducing ineffective work

Inactive Publication Date: 2016-11-23
PEKING UNIV SHENZHEN HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have a long detection period and are inflexible. They cannot find the problem of glass liquid compatibility in time, and it is easy to make follow-up detection invalid.

Method used

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  • Vitrification freezing observing device and application thereof
  • Vitrification freezing observing device and application thereof
  • Vitrification freezing observing device and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] See figure 1 , figure 2 and image 3 , The vitrification observation device includes a vertical microscope 1 , an isolation box 2 and a petri dish 3 . Upright microscope 1 comprises lens device 101, object stage 102 and the light source 103 below object stage 102, and upright microscope 1 is connected with photographing device, is used for observing and photographing vitrification---rethawing process and tissue in glass liquid balance.

[0042] The isolation box 2 is placed on the object stage 102 of the vertical microscope. The isolation box 2 is in the shape of a cuboid and is made of materials with ultra-low temperature resistance and thermal insulation properties. A hollow portion 21 is provided, and the hollow portion 21 is cylindrical, including a first hollow portion 211 and a second hollow portion 212 connected up and down, the diameter of the first hollow portion 211 is 9 centimeters, and the diameter at 1 centimeter from the surface of the isolation box 2 ...

Embodiment 2

[0046] Using a vitrification observation device to screen liquid glass includes the following steps:

[0047] (1) Rapid freezing observation of molten glass

[0048] A. Pour liquid nitrogen into the glass petri dish 3, and adjust the focal length of the vertical microscope 1;

[0049] B. Draw a certain volume of glass fluid to be tested with an ICSI injection needle 4 with an inner diameter of 0.5 microns, and place it in the marked focus position in the glass culture dish 3;

[0050] C. Photographing and recording the changes of the glass liquid, the observation indicators include the formation of crystals and fragments during the rapid freezing process;

[0051] (2) Observation of glass fluid rethawing

[0052] A. Pour 37°C water into the glass petri dish 3 to adjust the focal length of the vertical microscope 1;

[0053] B. Take out the above-mentioned ICSI injection needle 4 frozen in liquid nitrogen and place it in the marked focus position in the glass culture di...

Embodiment 3

[0061] The application of the vitrification observation device in the optimization of the equilibrium time of ovarian tissue in the glass liquid. Since the shape of the ovarian block used in the experiment is irregular, its volume change cannot be measured. Therefore, the change of the surface area of ​​the ovarian block can be observed in the glass liquid. penetration behavior. Said application includes the following steps:

[0062] (1) Add a certain amount of glass liquid into the glass petri dish 3;

[0063] (2) Place ovarian tissue in isotonic phosphate buffer;

[0064] (3) Move the ovarian tissue into the glass fluid, and osmotically balance for a certain period of time;

[0065] (4) Adjust the focal length of the vertical microscope 1, take pictures under the microscope every one minute, record the changes in the surface area of ​​the tissue, and measure the surface area with Image plus6.0 image analysis software;

[0066] (5) Taking the surface area of ​​ovarian tiss...

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Abstract

The invention discloses a vitrification freezing observing device. The vitrification freezing observing device comprises a vertical microscope, an isolation box and a culture dish; the vertical microscope is connected with a shooting device and used for observing and shooting; the hollow part of the isolation box is shaped like a cylinder, and the isolation box is arranged on a containing table of the vertical microscope and used for bearing observation materials and passing light rays below the containing table of the vertical microscope; the culture dish is arranged in the hollow part of the isolation box and used for containing the observation materials. When the vitrification freezing observing device is applied to optimal liquid glass screening and structure equilibrium time optimization, vitrification states of different liquid glass in the rapid freezing and remelting process and surface area changes of structure in the liquid glass in the equilibrium process can be conveniently observed, initial glassy state screening and optimization and determining of structure equilibrium time are completed, the advantages of being rapid, easy and convenient are achieved, and invalid work is greatly reduced.

Description

technical field [0001] The invention relates to a vitrification freezing observation device and its application in glass liquid screening and tissue equilibrium time optimization. Background technique [0002] The application value of tissue cryopreservation in the field of reproductive medicine has been recognized and some achievements have been made, but the progress is still slow. Among them, the vitrification method was first applied to the study of ovarian tissue cryopreservation in 2002. The principle is that high-concentration cryopreservation The protective solution can form a vitrified solution when the temperature is rapidly cooled, without the formation of intracellular and extracellular ice crystals, and is more suitable for cryopreservation of human ovarian tissues with complex structures. In 2013, the first baby was born after autotransplantation of vitrified ovarian tissue slices. The higher the concentration of the glass liquid, the easier it is to form a gla...

Claims

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Application Information

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IPC IPC(8): G01N21/84
Inventor 赵京卉曾玉翠汤惠茹吴瑞芳
Owner PEKING UNIV SHENZHEN HOSPITAL
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