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Use of In Situ Fourier Transform Infrared Spectroscopy in Cryobiological Research

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2180)

Abstract

In this chapter, we describe how Fourier transform infrared spectroscopy (FTIR) can be applied in cryobiological research to study: structure and thermal properties of biomolecules in cells and tissues, physical properties of cryopreservation and freeze-drying formulations, and permeation of molecules into cells and tissues. An infrared spectrum gives information about characteristic molecular vibrations of specific groups in molecules, whereas the temperature dependence of specific infrared bands may reveal information about conformational and phase changes. Infrared spectroscopy is minimally invasive and does not require labeling, whereas spectra can be recorded in any physical state of a sample. Data acquisition and spectral processing procedures are described to study phase state changes of protective formulations, cell membrane phase behavior during freezing and drying, protein denaturation during heating, and permeation of protective molecules into tissues. The latter can be used to estimate incubation times needed to load tissues with sufficient amounts of protective agents for cryopreservation or freeze-drying.

Key words

Dehydration Freeze-drying Freezing Diffusion Fourier transform infrared spectroscopy (FTIR) Membrane phase behavior Protein denaturation 

Notes

Acknowledgments

This work is supported by funding from the German Research Foundation (DFG, Deutsche Forschungsgemeinschaft) via grants WO1735/6-2 and SI1462/4-2.

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© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Unit for Reproductive Medicine—Clinic for HorsesUniversity of Veterinary Medicine HannoverHannoverGermany
  2. 2.Biostabilization Laboratory—Lower Saxony Centre for Biomedical Engineering, Implant Research and DevelopmentUniversity of Veterinary Medicine HannoverHannoverGermany

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