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

Emmanuel Beaurepaire 1 Thierry Boulesteix 1 G. Godeau 2 N. Pagès 3 Ana-Maria Pena 1 Martin-Pierre Sauviat 1 Marie-Claire Schanne-Klein 1 K. Senni 2
1 LOB - Laboratoire d'optique et biosciences
2 Paris V University, 1 rue Maurice-Arnoux, F-92120 Montrouge
3 Louis Pasteur University, 74 route du Rhin, F-67401 Illkirch Cedex
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. © (2005) COPYRIGHT SPIE-The International Society for Optical Engineering
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Emmanuel Beaurepaire, Thierry Boulesteix, G. Godeau, N. Pagès, Ana-Maria Pena, et al.. Multiphoton microscopy using intrinsic signals for pharmacological studies in unstained cardiac and vascular tissue. Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, Jan 2005, San Jose, CA, United States. pp.67, ⟨10.1117/12.589558⟩. ⟨hal-00844506⟩

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