title: Effects of tissue fixation on coherent anti-Stokes Raman scattering images of brain.
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.
authors.affiliation: Dresden University of Technology, Clinical Sensing and Monitoring, Faculty of Medicine, Fetscherstrasse 74, D-01307 Dresden, Germany.
authors.forename: Roberta
authors.initials: R
authors.lastname: Galli
authors.affiliation: Dresden University of Technology, Carl Gustav Carus University Hospital, Department of Neurosurgery, Fetscherstrasse 74, D-01307 Dresden, Germany.
authors.forename: Ortrud
authors.initials: O
authors.lastname: Uckermann
authors.affiliation: Dresden University of Technology, Clinical Sensing and Monitoring, Faculty of Medicine, Fetscherstrasse 74, D-01307 Dresden, Germany.
authors.forename: Edmund
authors.initials: E
authors.lastname: Koch
authors.affiliation: Dresden University of Technology, Carl Gustav Carus University Hospital, Department of Neurosurgery, Fetscherstrasse 74, D-01307 Dresden, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Schackert
authors.affiliation: Dresden University of Technology, Carl Gustav Carus University Hospital, Department of Neurosurgery, Fetscherstrasse 74, D-01307 Dresden, GermanycCenter for Regenerative Therapies Dresden, DFG Research Center and Cluster of Excellence, Fetscherstrasse 105, D-01307 Dresden, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Kirsch
authors.affiliation: Dresden University of Technology, Clinical Sensing and Monitoring, Faculty of Medicine, Fetscherstrasse 74, D-01307 Dresden, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Steiner
mesh_terms: D000096:Acetone / Q000737:chemistry; D000818:Animals; D001921:Brain / Q000473:pathology*; D001932:Brain Neoplasms / Q000473:pathology; D002490:Central Nervous System / Q000473:pathology; D005260:Female; D005557:Formaldehyde / Q000737:chemistry; D005909:Glioblastoma / Q000473:pathology; D006652:Histological Techniques; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008055:Lipids / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D051379:Mice; D008819:Mice, Nude; D013059:Spectrum Analysis, Raman; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D008055:Lipids; D000096:Acetone; D005557:Formaldehyde; D000432:Methanol