Scientific Results

Protein conformation and molecular order probed by second-harmonic-generation microscopy

Year: 2012

Authors: Vanzi F., Sacconi L., Cicchi R., Pavone FS.

Autors Affiliation: University of Florence, Department of Evolutionary Biology “Leo Pardi,” Florence, Italy; University of Florence, European Laboratory for Non-Linear Spectroscopy (LENS), Sesto Fiorentino, Italy; National Institute of Optics, National Research Council (INO-CNR), Florence, Italy; University of Florence, Department of Physics, Sesto Fiorentino, Italy

Abstract: Second-harmonic-generation (SHG) microscopy has emerged as a powerful tool to image unstained living tissues and probe their molecular and supramolecular organization. In this article, we review the physical basis of SHG, highlighting how coherent summation of second-harmonic response leads to the sensitivity of polarized SHG to the three-dimensional distribution of emitters within the focal volume. Based on the physical description of the process, we examine experimental applications for probing the molecular organization within a tissue and its alterations in response to different biomedically relevant conditions. We also describe the approach for obtaining information on molecular conformation based on SHG polarization anisotropy measurements and its application to the study of myosin conformation in different physiological states of muscle. The capability of coupling the advantages of nonlinear microscopy (micrometer-scale resolution in deep tissue) with tools for probing molecular structure in vivo renders SHG microscopy an extremely powerful tool for the advancement of biomedical optics, with particular regard to novel technologies for molecular diagnostic in vivo. (C) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE)

Journal/Review: JOURNAL OF BIOMEDICAL OPTICS

Volume: 17 (6)      Pages from: 060901  to: 060901

More Information: We thank Dr. Anna Letizia Allegra Mascaro for useful discussion about the manuscript. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement N. 228334 and Human Frontier Science Program research grant RGP0027/2009. This research project has been also supported by the Ente Cassa di Risparmio di Firenze (private foundation).
KeyWords: imaging; imaging coherence; microscopy; nonlinear optics; second-harmonic-generation
DOI: 10.1117/1.JBO.17.6.060901

Citations: 29
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English