Raman cryomicroscopic imaging and sample holder for spectroscopic subzero temperature measurements

Guanglin Yu; Rui Li; Allison Hubel

doi:10.1007/978-1-0716-0783-1_14

Raman cryomicroscopic imaging and sample holder for spectroscopic subzero temperature measurements

Guanglin Yu, Rui Li, Allison Hubel

Mechanical Engineering

Research output: Chapter in Book/Report/Conference proceeding › Chapter

2 Scopus citations

Abstract

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 language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	351-361
Number of pages	11
DOIs	https://doi.org/10.1007/978-1-0716-0783-1_14
State	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2180
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.

Keywords

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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10.1007/978-1-0716-0783-1_14

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title = "Raman cryomicroscopic imaging and sample holder for spectroscopic subzero temperature measurements",

abstract = "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.",

keywords = "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",

author = "Guanglin Yu and Rui Li and Allison Hubel",

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: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2021.",

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doi = "10.1007/978-1-0716-0783-1_14",

language = "English (US)",

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publisher = "Humana Press Inc.",

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AU - Yu, Guanglin

AU - Li, Rui

AU - Hubel, Allison

N1 - 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.

PY - 2021

Y1 - 2021

N2 - 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.

AB - 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.

KW - Cell

KW - Cryopreservation

KW - Hyperspectral imaging

KW - Low-temperature microscopy

KW - Raman spectroscopy

KW - Freeze Drying/methods

KW - Cold Temperature

KW - Jurkat Cells

KW - Cell Membrane/drug effects

KW - Humans

KW - Phase Transition

KW - Cryopreservation/methods

KW - Ice/analysis

KW - Image Processing, Computer-Assisted/methods

KW - Cryoprotective Agents/pharmacology

KW - Spectrum Analysis, Raman/methods

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M3 - Chapter

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T3 - Methods in Molecular Biology

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BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Raman cryomicroscopic imaging and sample holder for spectroscopic subzero temperature measurements

Abstract

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Bibliographical note

Keywords

PubMed: MeSH publication types

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