The Scanning Cryomacroscope - A Device Prototype for the Study of Cryopreservation.

Area of Science:

• Cryobiology
• Biophysics
• Microscopy

Background:

• In situ analysis of cryopreserved biological samples requires advanced visualization techniques.
• Previous cryomacroscope prototypes had limitations in sample size visualization and integrated cooling.

Purpose of the Study:

• To develop a novel scanning cryomacroscope prototype for enhanced in situ analysis of cryopreserved samples.
• To integrate a scanning mechanism for visualizing samples larger than the optical field of view.
• To ensure compatibility with existing cooling chambers, eliminating the need for dedicated cooling systems.

Main Methods:

• Development of a scanning cryomacroscope prototype with integrated scanning capabilities.
• Utilizing top-loading controlled-rate cooling chambers for sample preparation.
• Demonstrating visualization on cryoprotective agent (CPA) dimethyl sulfoxide and cryoprotective cocktail DP6.

Main Results:

• Successful integration of a scanning mechanism enabling visualization of larger samples.
• Demonstration of compatibility with standard cooling chambers.
• Visualization of key cryopreservation events including glass formation, crystallization, thermal contraction, and fracture.
• Generation of digital movies with overlaid experimental data.

Conclusions:

• The new scanning cryomacroscope offers significant improvements for in situ analysis of cryopreserved samples.
• The device facilitates comprehensive data capture and visualization of cryopreservation processes.
• This technology advances the field of cryobiology by providing a more versatile and integrated analytical tool.