Two-step temperature-reducing method for eliminating cryogenic fracture in biological tissue preserved by vitrification method
A vitrification and biological tissue technology, applied in the field of biomedicine, can solve the problems of low temperature fracture and the inability to continue cooling, and achieve the effects of easy operation of equipment, efficient vitrification preservation, and avoiding tissue fracture.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] This example is to investigate the breakage of the pure vitrification solution in the two-step cooling and recovery process in a water bath.
[0027] The two-step cooling process and devices are as follows: figure 1 with figure 2 shown. figure 2 Among them, the height of the Dewar bottle is 200mm, and the inner diameter is 105mm. The size of the cryovial is 2ml. The volume of the vitrification solution was 1 ml.
[0028] Pour liquid nitrogen with a height of about 40 mm into the Dewar bottle. For the sample position corresponding to the temperature range of -196 to -135 ° C in the liquid nitrogen gas phase at this height (also known as the gas phase distance, that is, the distance between the center of the sample and the liquid nitrogen The same below.) is 0-47.5mm, and the distance between the center of the vitrified solution in the freezing tube and the bottom of the tube is 7.5mm, so the thickness of the support plate should be 0-40mm. The thickness of the sup...
Embodiment 2
[0033] This embodiment is the determination of the operating parameters of the two-step temperature reduction.
[0034] In order to determine the operating parameters of the two-step cooling process, at first investigated the fracture phenomenon that the liquid nitrogen depth is 40mm and the gas phase distance value is 17.5mm, 27.5mm, 37.5mm, 47.5mm and 57.5mm respectively, the method is the same as in Example 1; then measure In the above cases, the temperature change of the vitrification solution is processed to obtain the change of the cooling rate; the experimental phenomenon results and the cooling rate measurement results are combined to finally determine the range of operating parameters allowed to eliminate the main fracture of the solution.
[0035]In this experiment, the volume of the solution used was 1.0 ml, so the solution depth was 15 mm. Therefore, when the thickness of the support plate is 10mm, the gas phase distance of the cryopreservation system is 17.5mm. B...
Embodiment 3
[0048] This example is to investigate the fracture situation of three kinds of tissues after two-step cooling and recovery in water bath.
[0049] Use a corneal trephine to drill a Φ8mm tissue block in the center of the bovine cornea; cut a 5mm×5mm×8mm tissue block from the pig’s rib cartilage; cut out a bovine artery with a size of 5mm×8mm×2mm, of which 2mm is the vessel wall thickness of. The three kinds of tissues were respectively soaked in the vitrification solution to balance the penetration of the protective agent.
[0050] The fracture phenomena of the tissue were observed macroscopically and microscopically. Microscopic observation is to observe whether there is a fracture on the cornea under an inverted optical microscope at 200 times. At the same time, the traditional one-step cooling and rewarming treatment was also performed on the tissue, and the fracture was observed; the tissue that was not equilibrated and frozen with the protective agent was used as a contr...
PUM
| Property | Measurement | Unit |
|---|---|---|
| thickness | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More