4.5 Article

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

期刊

JOURNAL OF BIOMEDICAL OPTICS
卷 17, 期 6, 页码 -

出版社

SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.JBO.17.6.060901

关键词

imaging; imaging coherence; microscopy; nonlinear optics; second-harmonic-generation; tissues

资金

  1. European Union [228334]
  2. Human Frontier Science Program research grant [RGP0027/2009]
  3. Ente Cassa di Risparmio di Firenze (private foundation)

向作者/读者索取更多资源

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). [DOI: 10.1117/1.JBO.17.6.060901]

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