Supplemented phase diagrams for vitrification CPA cocktails: DP6, VS55 and M22

Zonghu Han, Lakshya Gangwar, E. Magnuson, M. L. Etheridge, C. O. Pringle, J. C. Bischof, J. Choi

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


DP6, VS55 and M22 are the most commonly used cryoprotective agent (CPA) cocktails for vitrification experiments in tissues and organs. However, complete phase diagrams for the three CPAs are often unavailable or incomplete (only available for full strength CPAs) thereby hampering optimization of vitrification and rewarming procedures. In this paper, we used differential scanning calorimetry (DSC) to measure the transition temperatures including heterogeneous nucleation temperatures (Thet), glass transition temperatures (Tg), rewarming phase crystallization (devitrification and/or recrystallization) temperatures (Td) and melting temperatures (Tm) while cooling or warming the CPA sample at 5 °C/min and plotted the obtained transition temperatures for different concentrations of CPAs into the phase diagrams. We also used cryomicroscopy cooling or warming the sample at the same rate to record the ice crystallization during the whole process, and we presented the cryomicroscopic images at the transition temperatures, which agreed with the DSC presented phenomena.

Original languageEnglish (US)
Pages (from-to)113-121
Number of pages9
StatePublished - Jun 2022

Bibliographical note

Funding Information:
This work was supported by NIH R01DK117425 and R01HL135046 and NSF EEC 1941543 .

Publisher Copyright:
© 2022 Elsevier Inc.


  • DP6
  • M22
  • Phase diagram
  • Vitrification
  • VS55

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.


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