Raman cryomicroscopic imaging and sample holder for spectroscopic subzero temperature measurements

Guanglin Yu, Rui Li, Allison Hubel

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations


Raman spectroscopy has been gaining in popularity for noninvasive analysis of single cells. Raman spectra and images deliver meaningful information regarding the biochemical, biophysical, and structural properties of cells in various states. Low-temperature Raman spectroscopy has been applied to verify the presence of ice inside a frozen cell and to illustrate the distribution of both penetrating and non-penetrating cryoprotectants. This chapter delineates Raman cryomicroscopic imaging of single cells as well as sample handling for spectroscopic measurements at subzero temperature. The experimental setup is depicted with a special emphasis on a custom-built temperature-controlled cooling stage. The use of Raman cryomicroscopic imaging is demonstrated using Jurkat cells cryopreserved in a sucrose solution. Moreover, strategies for determining intracellular ice formation (IIF) and analysis of sucrose partitioning across the cell membrane are presented.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages11
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
This work was partially supported by funding from the National Institute of Biomedical Imaging and Bioengineering of the National Institutes of Health under award number R01EB023880. Raman spectroscopy in this demonstration was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the National Science Foundation through the MRSEC program.

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2021.


  • Cell
  • Cryopreservation
  • Hyperspectral imaging
  • Low-temperature microscopy
  • Raman spectroscopy
  • Freeze Drying/methods
  • Cold Temperature
  • Jurkat Cells
  • Cell Membrane/drug effects
  • Humans
  • Phase Transition
  • Cryopreservation/methods
  • Ice/analysis
  • Image Processing, Computer-Assisted/methods
  • Cryoprotective Agents/pharmacology
  • Spectrum Analysis, Raman/methods

PubMed: MeSH publication types

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


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