Author(s)

Dominique Dumas, Elisabeth Werkmeister, Sebastien Hupont, Céline Huselstein, Natalia De Isla, Marthe Rousseau, Patrick Menu, Didier Mainard

Ingénierie Moléculaire et Thérapeutique (CNRS FR 3209), Plate Forme d’Imagerie Cellulaire et Tissulaire PTIBC-IBISA NANCY, Nancy, France.
Plate-Forme Microscopie-Imagerie-Cytométrie du campus Pasteur Lille (MICPaL), IFR 142 Institut Pasteur de Lille, Institut de Biologie de Lille, Lille, France.
Universite de Lorraine, Physiopathologie, Pharmacologie et Ingénierie Articulaires (IMOPA, CNRS 7365), Nancy, France.
Universite de Lorraine, Physiopathologie, Pharmacologie et Ingénierie Articulaires (IMOPA, CNRS 7365), Nancy, France; Ingénierie Moléculaire et Thérapeutique (CNRS FR 3209), Plate Forme d’Imagerie Cellulaire et Tissulaire PTIBC-IBISA NANCY, Nancy, France.

In this work, we have considered a new multimodality imaging for macroscopy based on Second Harmonic Generation (SHG) method to monitor invasivelessly the matrix collagen. As the triple helicoidally structure of collagen molecules appearing as not centrosymetric, very organized and spatially oriented, collagen fibrils give rise to a very strong SHG signal and can be imaged without any exogenous dye. To integrate a multidimensional scale with a large field of view (non-sliced samples), we have adapted and validated an instrumental coupling between a two photon excitation laser and a macroscope to collect cartography of SHG signal. We introduced an index (F-SHG) based on decay time response measured by TCSPC for respectively Fluorescence (F) and Second Harmonic Generation (SHG) values. For various sample where protein collagen is the major component of extracellular matrix (vessel, skin, carotide vessel, rat femoral head cartilage, mouse tumor, human wharton’s jelly and rat tendon) or not (nacre), we compared the index distribution obtained with MacroSHG. In this work, we showed for the first time that multiscale large field imaging (Macroscopy) combined to Multimodality approaches (SHG-TCSPC) could be an innovative and non-invasive technique to detect and identify some biological interest molecules (collagen) in biomedical topics.

Multiphoton Macroscopy, Second Harmonic Generation, Collagen, Extracellular Matrix

Dumas, D. , Werkmeister, E. , Hupont, S. , Huselstein, C. , Isla, N. , Rousseau, M. , Menu, P. and Mainard, D. (2014) Non-Invasive Second Harmonic Generation (SHG) in Macroscopy (MacroSHG) as Bio-Diagnosis to Image Collagen Network Organization in Extracellular Matrix. Engineering, 6, 485-490. doi: 10.4236/eng.2014.68050.