Quantifying the Growth Rate and Morphology of Ice Crystals Growing in Cryoprotectants via High-Speed Camera and Cryo-Microscope

2013 ◽  
Author(s):  
Xu Xue ◽  
Zhi Zhu He ◽  
Jing Liu
Keyword(s):  
Growth Rate ◽  
Cooling Rate ◽  
High Speed ◽  
Ice Crystals ◽  
Experimental Methodology ◽  
Impact Factors ◽  
High Speed Camera ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Near Future

Recently, several significant progresses have been made on the studies of extracellular and intracellular ice formation based on high-speed camera and cryomicroscope. This experimental methodology could accurately capture rapid formation process of ice crystals at micro-scale. In this paper, we are dedicated to quantify and comparatively investigate the growth rate and morphology of ice crystals growing in DMSO, sucrose and trehalose, respectively via high-speed camera and cryo-microscope. Several impact factors such as the concentration of cryoprotectants and the cooling rate have been investigated. The results indicate that the species and concentration of cryoprotectants and the cooling rate could significantly affect the growth rate and morphology of ice crystals. DMSO is better than trehalose and sucrose as cryoprotectant because of its molecular structure. This work may enhance current understanding of the factors for ice crystals formation and help optimize the cryopreservation process in the near future.

Quantifying the Growth Rate and Morphology of Ice Crystals Growth in Cryoprotectants Via High-Speed Camera and Cryomicroscope

2015 ◽  
Vol 137 (9) ◽  
Author(s):  
Xu Xue ◽  
Hai-Lan Jin ◽  
Zhi-Zhu He ◽  
Jing Liu
Keyword(s):  
Growth Rate ◽  
Cooling Rate ◽  
High Speed ◽  
Ice Crystals ◽  
Experimental Methodology ◽  
Impact Factors ◽  
High Speed Camera ◽  
Change Behavior ◽  
Data Processing Method ◽  
Intracellular Ice Formation

Recently, several significant progresses have been made on the studies of extracellular and intracellular ice formation based on high-speed camera and cryomicroscope. This experimental methodology could accurately capture the rapid formation process of ice crystals at microscale. However, quantitative interpretation on such phase change behavior still reserved a tough issue. Here, in this paper, we quantitatively studied the ice crystals growth in three kinds of cryoprotectants like dimethyl sulfoxide (DMSO), sucrose, and trehalose via high-speed camera, cryomicroscope as well as the proposed data processing method. Several critical impact factors such as the concentration of cryoprotectants and the cooling rate have been investigated. Particularly, an efficient image processing technology has been developed to quantify the growth rate and morphology of the ice crystals. The results indicate that the species and concentration of cryoprotectants and the cooling rate could significantly affect the growth rate and morphology of ice crystals. DMSO is better than trehalose and sucrose as cryoprotectant because of the molecular structure. This work established a new methodology to quantify the ice crystals growth and would enhance current understanding of the factors for ice crystals formation. It is also expected to help optimize the cryopreservation process in the near future.

A Simple Cryomicroscopic Method to Measure the Size of Intracellular Ice Crystals

2009 ◽  
Author(s):  
Xu Han ◽  
Hongbin Ma ◽  
John K. Critser
Keyword(s):  
Melting Point ◽  
Ethylene Glycol ◽  
Cooling Rate ◽  
Ice Crystals ◽  
Ice Formation ◽  
Melting Point Depression ◽  
Critical Importance ◽  
Factors Influencing ◽  
Intracellular Ice Formation ◽  
Intracellular Ice

Investigating the factors influencing the characteristics of intracellular ice formation (IIF) is of critical importance for cryopreservation and cryosurgery techniques. However, for the detection of the size of intracellular ice crystals, ∼10nm-0.1μm, there exist serious technical and theoretical difficulties. In this study, a cryomicroscopic method was established to measure the size of intracellular ice crystals in mouse oocytes during their warming processes by investigating the melting point depression of the intracellular ice crystals from extracellular ones. Using the Gibbs-Thomson relation, the size of intracellular ice crystals was calculated and the results range from 4–28 nm, when the molality of the extracellular ethylene glycol and NaCl ranges from 0 to 4m and 0.15 to 0.6m, respectively, and the cooling rate is 100K/min.

Small-volume vitrification and rapid warming yield high survivals of one-cell rat embryos in cryotubes

2021 ◽  
Author(s):  
Yasuyoshi Fukuda ◽  
Misako Higashiya ◽  
Takahiro Obata ◽  
Keita Basaki ◽  
Megumi Yano ◽  
...  
Keyword(s):  
Cooling Rate ◽  
Small Volume ◽  
Sucrose Solution ◽  
High Concentration ◽  
Rat Embryos ◽  
Low Concentrations ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Warming Rate ◽  
Very High

Abstract To cryopreserve cells, it is essential to avoid intracellular ice formation during cooling and warming. One way to achieve this is to convert the water inside the cells into a non-crystalline glass. It is currently believed that to accomplish this vitrification, the cells must be suspended in a very high concentration (20–40%) of a glass-inducing solute, and subsequently cooled very rapidly. Herein, we report that this belief is erroneous with respect to the vitrification of one-cell rat embryos. In the present study, one-cell rat embryos were vitrified with 5 μL of EFS10 (a mixture of 10% ethylene glycol, 27% Ficoll, and 0.45 M sucrose) in cryotubes at a moderate cooling rate, and warmed at various rates. Survival was assessed according to the ability of the cells to develop into blastocysts and to develop to term. When embryos were vitrified at a 2,613 °C/min cooling rate and thawed by adding 1 mL of sucrose solution (0.3 M, 50 °C) at a warming rate of 18,467 °C/min, 58.1 ± 3.5% of the EFS10-vitrified embryos developed into blastocysts, and 50.0 ± 4.7% developed to term. These rates were similar to those of non-treated intact embryos. Using a conventional cryotube, we achieved developmental capabilities in one-cell rat embryos by rapid warming that were comparable to those of intact embryos, even using low concentrations (10%) of cell-permeating cryoprotectant and at low cooling rates.

The effect of various cooling rates on the membrane ultrastructure of frozen human erythrocytes and its relation to the extent of haemolysis after thawing

1981 ◽  
Vol 49 (1) ◽  
pp. 369-382
Author(s):  
S. Fujikawa
Keyword(s):  
Cooling Rate ◽  
Human Erythrocytes ◽  
Ice Crystals ◽  
Erythrocyte Membranes ◽  
Ultrastructural Changes ◽  
Cooling Rates ◽  
Intracellular Ice ◽  
Frozen State ◽  
Membrane Ultrastructure ◽  
Fast Freezing

Human erythrocytes suspended in buffered isotonic saline were frozen to the temperature of liquid nitrogen at various cooling rates of 3, 140, 700, 1800, 3500, 8000 and 11 500 deg. C/min. The membrane ultrastructure in the frozen state and the extent of haemolysis after thawing were examined at each cooling rate. As the cooling rates increased from 3 to 3500 deg. C/min, the extent of lysis gradually decreased, but further increase in cooling rates in excess of 8000 deg. C/min resulted in an abrupt increase of lysis. Membrane-associated vesicles devoid of intramembrane particles (IMPs) were formed in the erythrocyte membranes frozen at cooling rates slower than 1800 deg. C/min. The frequency and size of these vesicles were highly cooling-rate-dependent and they were no longer formed in the erythrocyte membranes frozen at cooling rates faster than 3500 deg. C/min. Another membrane ultrastructural change associated closely with the formation of intracellular ice crystals appeared at cooling rates faster than 8000 deg. C/min. The membrane regions in direct contact with intracellular ice crystals were physically damaged and had an appearance resembling worm-eaten spots. The erythrocytes frozen at a cooling rate of 3500 deg. C/min exhibited ultrastructural integrity of the membrane by avoiding the membrane changes caused by either slow or fast freezing. It is suggested, from the close relation between membrane ultrastructure and the extent of haemolysis, that the ultrastructural integrity of membrane in the frozen state is important for avoiding haemolysis after thawing, and that the membrane ultrastructural changes caused by both slow and fast freezing were responsible for the lysis after thawing.

Analysis of Cracking Processes of Sandstone in Rockburst Test

2012 ◽  
Vol 256-259 ◽  
pp. 1052-1059
Author(s):  
De Jian Li ◽  
Fei Zhao ◽  
Run Jie Ma ◽  
Li Qiang Han ◽  
Xiao Hu Wu
Keyword(s):  
Growth Rate ◽  
Fractal Dimension ◽  
Crack Propagation ◽  
High Speed ◽  
Fractal Dimensions ◽  
Rock Surface ◽  
Horizontal Line ◽  
High Speed Camera ◽  
Camera System ◽  
Very High

High speed camera system is used to record crack propagation on rock surface during the lab rocburst experiment. This paper presents the specific analytical method of crack propagation with high speed images. Crack propagation types and directions are briefly described, and the lengths, growth rate and fractal dimension of cracks are calculated. The results show that the crack grows mainly along the horizontal line on the unloading surface, at a low speed before rockburst and reaches a very high speed abruptly near the burst point. The fractal dimensions demonstrated that the crack turns to be rough when rockburst happens. The study on the crack propagation is benefit to the understanding on the mechanism of rockburst.

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