Abstract
An infrared spectrum gives information about characteristic molecular vibrations of specific groups in molecules. Fourier transform infrared spectroscopy can be applied to study lipids and proteins in cells or tissues. Spectra can be collected during cooling, heating, or dehydration of a sample using a temperature-controlled sample holder or a sample holder for controlled dehydration. In the current chapter, acquisition and analysis of infrared spectra during cooling, warming, or dehydration is described. Spectra analysis involving assessment of specific band positions, areas, or ratios is described. Special emphasis is given on studying membrane phase behavior and protein denaturation in cells or tissues. In addition, methods are presented to determine the water-to-ice phase change during freezing, dehydration kinetics, and the glass transition temperature of amorphous systems.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Wolkers WF, Hoekstra FA (2003) In situ FTIR assessment of desiccation-tolerant tissues. Spectroscopy 17:297–313
Wolkers WF, Oldenhof H (2010) In situ FTIR studies on mammalian cells. Spectroscopy 24:525–534
Wolkers WF, Balasubramanian SK, Ongstad EL, Zec H, Bischof JC (2007) Effects of freezing on membranes and proteins in LNCaP prostate tumor cells. Biochim Biophys Acta 1768:728–736
Wolkers WF, Oldenhof H, Glasmacher B (2010) Effect of trehalose on dehydration kinetics of phospholipid vesicles, as measured in real time using ATR infrared spectroscopy. Cryobiology 61:108–114
Mantsch HH, McElhaney RN (1991) Phospholipid phase transitions in model and biological membranes as studied by infrared spectroscopy. Chem Phys Lipids 57:213–226
Goormaghtigh E, Cabiaux V, Ruysschaert JM (1994) Determination of soluble and membrane protein structure by Fourier transform infrared spectroscopy. I. Assignments and model compounds. Subcell Biochem 23:329–362
Wolkers WF, Oldenhof H, Alberda M, Hoekstra FA (1998) A Fourier transform infrared microspectroscopy study of sugar glasses: application to anhydrobiotic higher plant cells. Biochim Biophys Acta 1379:83–96
O’Brien FEM (1948) The control of humidity by saturated salt solutions. J Sci Instrum 25:73–76
Akhoondi M, Oldenhof H, Sieme H, Wolkers WF (2012) Freezing-induced removal of water from phospholipid head groups in biomembranes. BSI 1:293–302
Oldenhof H, Akhoondi M, Sieme H, Wolkers WF (2013) Use of Fourier transform infrared spectroscopy to study membrane properties of cells at subzero temperatures. BSI 2:83–90
Acknowledgment
This work is supported by funding from the German Research Foundation (DFG, Deutsche Forschungsgemeinschaft) for the Cluster of Excellence “From Regenerative Biology to Reconstructive Therapy” (REBIRTH).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Wolkers, W.F., Oldenhof, H. (2015). Use of In Situ Fourier Transform Infrared Spectroscopy to Study Freezing and Drying of Cells. In: Wolkers, W., Oldenhof, H. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology, vol 1257. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-2193-5_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2193-5_5
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4939-2192-8
Online ISBN: 978-1-4939-2193-5
eBook Packages: Springer Protocols