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Label-Free Live-Cell Imaging with Confocal Raman Microscopy

Klein, Katharina and Gigler, Alexander M. and Aschenbrenner, Thomas and Monetti, Roberto and Bunk, Wolfram and Jamitzky, Ferdinand and Morfill, Gregor and Stark, Robert W. and Schlegel, Jürgen (2012):
Label-Free Live-Cell Imaging with Confocal Raman Microscopy.
In: Biophysical Journal, Elsevier Science Publishing, pp. 360-368, 102, (2), ISSN 00063495, [Online-Edition: http://dx.doi.org/10.1016/j.bpj.2011.12.027],
[Article]

Abstract

Confocal Raman spectroscopy is a noninvasive alternative to established cell imaging methods because it does not require chemical fixation, the use of fluorescent markers, or genetic engineering. In particular, single live-cell, high-resolution imaging by confocal Raman microscopy is desirable because it allows further experiments concerning the individually investigated cells. However, to derive meaningful images from the spectroscopic data, one must identify cell components within the dataset. Using immunofluorescence images as a reference, we derive Raman spectral signatures by means of information measures to identify cell components such as the nucleus, the endoplasmic reticulum, the Golgi apparatus, and mitochondria. The extracted signatures allow us to generate representations equivalent to conventional (immuno)fluorescence images with more than three cell components at a time, exploiting the Raman spectral information alone.

Item Type: Article
Erschienen: 2012
Creators: Klein, Katharina and Gigler, Alexander M. and Aschenbrenner, Thomas and Monetti, Roberto and Bunk, Wolfram and Jamitzky, Ferdinand and Morfill, Gregor and Stark, Robert W. and Schlegel, Jürgen
Title: Label-Free Live-Cell Imaging with Confocal Raman Microscopy
Language: English
Abstract:

Confocal Raman spectroscopy is a noninvasive alternative to established cell imaging methods because it does not require chemical fixation, the use of fluorescent markers, or genetic engineering. In particular, single live-cell, high-resolution imaging by confocal Raman microscopy is desirable because it allows further experiments concerning the individually investigated cells. However, to derive meaningful images from the spectroscopic data, one must identify cell components within the dataset. Using immunofluorescence images as a reference, we derive Raman spectral signatures by means of information measures to identify cell components such as the nucleus, the endoplasmic reticulum, the Golgi apparatus, and mitochondria. The extracted signatures allow us to generate representations equivalent to conventional (immuno)fluorescence images with more than three cell components at a time, exploiting the Raman spectral information alone.

Journal or Publication Title: Biophysical Journal
Volume: 102
Number: 2
Publisher: Elsevier Science Publishing
Divisions: 11 Department of Materials and Earth Sciences
11 Department of Materials and Earth Sciences > Material Science
11 Department of Materials and Earth Sciences > Material Science > Physics of Surfaces
DFG-Collaborative Research Centres (incl. Transregio)
DFG-Collaborative Research Centres (incl. Transregio) > Collaborative Research Centres
Exzellenzinitiative
Exzellenzinitiative > Clusters of Excellence
Zentrale Einrichtungen
Exzellenzinitiative > Clusters of Excellence > Center of Smart Interfaces (CSI)
Date Deposited: 16 Jun 2014 09:31
Official URL: http://dx.doi.org/10.1016/j.bpj.2011.12.027
Identification Number: doi:10.1016/j.bpj.2011.12.027
Funders: This work was supported by the Deutsche Forschungsgemeinschaft Cluster of Excellence “Nanosystems Initiative Munich” and grant DFG-SFB-824 (Project B6: Imaging for Selection, Monitoring and Individualization of Cancer Therapies).
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