Special Section on Selected Topics in Biophotonics: Optical Coherence Tomography and Biomolecular Imaging with Coherent Raman Scattering Microscopy

Effects of tissue fixation on coherent anti-Stokes Raman scattering images of brain

[+] Author Affiliations
Roberta Galli, Edmund Koch, Gerald Steiner

Dresden University of Technology, Clinical Sensing and Monitoring, Faculty of Medicine, Fetscherstrasse 74, D-01307 Dresden, Germany

Ortrud Uckermann, Gabriele Schackert

Dresden University of Technology, Carl Gustav Carus University Hospital, Department of Neurosurgery, Fetscherstrasse 74, D-01307 Dresden, Germany

Matthias Kirsch

Dresden University of Technology, Carl Gustav Carus University Hospital, Department of Neurosurgery, Fetscherstrasse 74, D-01307 Dresden, Germany

Center for Regenerative Therapies Dresden, DFG Research Center and Cluster of Excellence, Fetscherstrasse 105, D-01307 Dresden, Germany

J. Biomed. Opt. 19(7), 071402 (Dec 23, 2013). doi:10.1117/1.JBO.19.7.071402
History: Received July 30, 2013; Revised November 5, 2013; Accepted November 14, 2013
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Abstract.  Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging multiphoton technique for the label-free histopathology of the central nervous system, by imaging the lipid content within the tissue. In order to apply the technique on standard histology sections, it is important to know the effects of tissue fixation on the CARS image. Here, we report the effects of two common fixation methods, namely with formalin and methanol–acetone, on mouse brain and human glioblastoma tissue. The variations induced by fixation on the CARS contrast and intensity were compared and interpreted using Raman microspectroscopy. The results show that, whenever unfixed cryosections cannot be used, fixation with formalin constitutes an alternative which does not deteriorate substantially the contrast generated by the different brain structures in the CARS image. Fixation with methanol–acetone strongly modifies the tissue lipid content and is therefore incompatible with the CARS imaging.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Roberta Galli ; Ortrud Uckermann ; Edmund Koch ; Gabriele Schackert ; Matthias Kirsch, et al.
"Effects of tissue fixation on coherent anti-Stokes Raman scattering images of brain", J. Biomed. Opt. 19(7), 071402 (Dec 23, 2013). ; http://dx.doi.org/10.1117/1.JBO.19.7.071402


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