Direct Comparison of Isobaric and Isochoric Vitrification of Two Aqueous Solutions with Photon Counting X-Ray Computed Tomography

TL;DR

This study uses photon counting X-ray CT to compare vitrification and ice formation in isochoric and isobaric conditions, revealing similar ice volumes but different cavity morphologies in confined aqueous solutions.

Contribution

It introduces photon counting X-ray CT for analyzing vitrification in confined systems and compares isochoric and isobaric conditions with controlled boundary conditions.

Findings

Similar ice and vapor volume fractions in both conditions

Distinct cavity morphologies in isochoric systems

Vapor outgassing or cavitation observed in confined cooling

Abstract

Vitrification is a promising approach for ice-free cryopreservation of biological material, but progress is hindered by the limited set of experimental tools for studying processes in the interior of the vitrified matter. Isochoric cryopreservation chambers are often metallic, and their opacity prevents direct visual observation. In this study, we introduce photon counting X-ray computed tomography (CT) to compare the effects of rigid isochoric and unconfined isobaric conditions on vitrification and ice formation during cooling of two aqueous solutions: 50 wt% DMSO and a coral vitrification solution, CVS1. Previous studies have only compared vitrification in isochoric systems with isobaric systems that have an exposed air-liquid interface. We use a movable piston to replicate the surface and thermal boundary conditions of the isochoric system yet maintain isobaric conditions. When controlling for the boundary conditions we find that similar ice and vapor volume fractions form during cooling in isochoric and isobaric conditions. Interestingly, we observe distinct ice and vapor cavity morphology in the isochoric systems, possibly due to vapor outgassing or cavitation as rapid cooling causes the pressure to drop in the confined systems. These observations highlight the array of thermo-fluidic processes that occur during vitrification in confined aqueous systems and motivate the further application of imaging techniques such as photon counting X-ray CT in fundamental studies of vitrification.