Droplet freeze-thawing system based on solid surface vitrification and laser rewarming.
10.7507/1001-5515.202305004
- Author:
Wenxin ZHU
1
;
Ping'an PAN
1
;
Yonghua HUANG
1
;
Wei CHEN
2
;
Sha HAN
2
;
Zheng LI
2
;
Jinsheng CHENG
3
Author Information
1. Institute of Refrigeration and Cryogenics, Shanghai Jiao Tong University, Shanghai 200240, P. R. China.
2. Department of Andrology, Center for Men's Health, Institute of Urology, Urologic Medical Center, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200080, P. R. China.
3. Shanghai Tofflon Medical Equipment Co., Ltd, Shanghai 201108, P. R. China.
- Publication Type:Journal Article
- Keywords:
Cell cryopreservation;
Laser warming;
Solid surface vitrification;
Vitrification
- MeSH:
Freezing;
Vitrification;
Cryopreservation/methods*;
Trehalose;
Gold;
Rewarming;
Metal Nanoparticles;
Cryoprotective Agents;
Lasers
- From:
Journal of Biomedical Engineering
2023;40(5):973-981
- CountryChina
- Language:Chinese
-
Abstract:
Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 μL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 3 °C/min. The volume range of 1-8 μL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 6 °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
