title: Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections.
abstract: Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a destaining protocol. Our results reveal that bringing tissue sections to a physiological state yields a significant improvement in nonlinear signals, particularly in SHG. Additionally, the destaining of sections previously processed with H&E staining significantly improves their SHG emission signals during imaging, thereby allowing sufficient analysis of collagen in these sections. These results are important for researchers and pathologists to obtain additional information from paraffin-embedded tissues and archived samples to perform retrospective analysis of the ECM or gain additional information from rare samples.
authors.affiliation: 1 Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University Tübingen , Tübingen, Germany .
authors.forename: Michael G
authors.initials: MG
authors.lastname: Monaghan
authors.affiliation: 1 Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University Tübingen , Tübingen, Germany .
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kroll
authors.affiliation: 1 Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University Tübingen , Tübingen, Germany .
authors.forename: Sara Y
authors.initials: SY
authors.lastname: Brucker
authors.affiliation: 1 Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University Tübingen , Tübingen, Germany .|2 Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Stuttgart, Germany .|3 Department of Medicine/Cardiology, Cardiovascular Research Laboratories, University of California , Los Angeles, California.
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
mesh_terms: D000818:Animals; D001021:Aortic Valve / Q000033:anatomy & histology* / Q000378:metabolism; D003094:Collagen / Q000032:analysis*; D005109:Extracellular Matrix / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods*; D011664:Pulmonary Valve / Q000033:anatomy & histology* / Q000378:metabolism; D013552:Swine
chemical_list: D003094:Collagen