Abstract
A new cryomacroscope prototype—a visualization device for the in situ analysis of cryopreserved biological samples—is presented in the current study. In order to visualize samples larger than the field of view of the optical setup, a scanning mechanism is integrated into the system, which represents a key improvement over previous cryomacroscope prototypes. Another key feature of the new design is in its compatibility with available top-loading controlled-rate cooling chambers, which eliminates the need for a dedicated cooling mechanism. The objective for the current development is to create means to generate a single digital movie of an experimental investigation, with all relevant data overlaid. The visualization capabilities of the scanning cryomacroscope are demonstrated in the current study on the cryoprotective agent dimethyl sulfoxide and the cryoprotective cocktail DP6. Demonstrated effects include glass formation, various regimes of crystallization, thermal contraction, and fracture formation.
Highlights
Controlling ice formation is key to the success of cryopreservation—the preservation of tissues at very low temperatures
For the purpose of evaluating the new device, the current study focuses on physical events occurring during cooling of a dimethyl sulfoxide solution (DMSO) at various concentrations and the cryoprotective cocktail DP6, which is a mixture of 234.4 g/L DMSO (3 M), 228.3 g/L propylene glycol (3 M), and 2.4 g/L HEPES in a Euro-Collins solution
A new cryomacroscope prototype is presented in the current study for the analysis of cryopreservation by vitrification in large-size samples
Summary
Controlling ice formation is key to the success of cryopreservation—the preservation of tissues at very low temperatures. Thermal expansion may be driven solely by molecular effects in any given phase of state [8,9], may result from phase transition (for example, pure water changes volume by about 9% upon freezing), or may result from glass transition effects [10,11]. It is not the thermal expansion per se but the constraint of thermal expansion that gives rise to thermal stress [12,13]
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