Multiphoton microscopy using intrinsic signals for pharmacological studies in unstained cardiac and vascular tissue

Emmanuel Beaurepaire; Thierry Boulesteix; Ana-Maria Pena; Nicole Pages; Karim Senni; Gaston Godeau; Martin-Pierre Sauviat; Marie-Claire Schanne-Klein

doi:10.1117/12.589558

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29 March 2005 Multiphoton microscopy using intrinsic signals for pharmacological studies in unstained cardiac and vascular tissue

Emmanuel Beaurepaire, Thierry Boulesteix, Ana-Maria Pena, Nicole Pages, Karim Senni, Gaston Godeau, Martin-Pierre Sauviat, Marie-Claire Schanne-Klein

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Proceedings Volume 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III; (2005) https://doi.org/10.1117/12.589558
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

We report two novel applications of multiphoton microscopy for pharmacological studies of unstained cardiovascular tissue. First, we show that second harmonic generation (SHG) microscopy of unstained cardiac myocytes can be used to determine the sarcomere length with sub-resolution accuracy, owing to the remarkable contrast of the SHG signal originating from myosin filaments. A measurement precision of 20 nm is achieved, taking the sample variability into account. We used this technique to measure sarcomere contracture in the presence of saxitoxin, and results were in agreement with mechanical measurements of atrial tissue contracture. Second, we characterized multiphoton microscopy of intact unlabeled arteries. We performed simultaneous detection of two-photon-excited fluorescence (2PEF) from elastin laminae and SHG from collagen fibers upon 860 nm excitation. Combined 2PEF/SHG images provide a highly specific, micron scale description of the architecture of these two major components of the vessel wall. We used this methodology to study the effects of lindane (a pesticide) on the artery wall structure and evidenced structural alteration of the vessel morphology.

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Emmanuel Beaurepaire, Thierry Boulesteix, Ana-Maria Pena, Nicole Pages, Karim Senni, Gaston Godeau, Martin-Pierre Sauviat, and Marie-Claire Schanne-Klein "Multiphoton microscopy using intrinsic signals for pharmacological studies in unstained cardiac and vascular tissue", Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); https://doi.org/10.1117/12.589558

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