Multi-view cryomacroscopy: A new device for study of cryopreservation by vitrification

Area of Science:

• Cryopreservation and Vitrification Science
• Biophysics and Material Science

Background:

• Cryomacroscopy has evolved over two decades to observe physical phenomena during cryopreservation.
• Previous devices had limitations in studying large-volume vitrification.
• Modeling has been used to infer material properties from cryomacroscopic observations.

Purpose of the Study:

• To present an innovative multi-view cryomacroscope (MVC) for studying large-sample vitrification.
• To address limitations of previous cryomacroscopes.
• To demonstrate the MVC's capability in observing physical effects during vitrification.

Main Methods:

• Development of a standalone thermo-optical device, the MVC.
• Integration of up to 12 cameras for multi-angle observation.
• Demonstration of MVC operation on the vitrification of a 60 ml cryoprotective agent (CPA) sample.

Main Results:

• Observed crystallization at the CPA-air interface during cooling.
• Documented fracturing within the vitrified CPA.
• Identified air suction artifact into the CPA center via thermocouple wires, causing bubbles.
• Observed rewarming phase crystallization (RPC).

Conclusions:

• The MVC is crucial for observing and understanding physical phenomena in large-sample vitrification.
• Preventing air suction artifacts is important to avoid adverse effects on cooling, crystallization, and fracturing.
• The MVC highlights the ongoing need for advanced cryomacroscopy in vitrification research.