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Formation of Ordered Biomolecular Structures by the Self-assembly of Short Peptides

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

Francesco Vanzi¹, Leonardo Sacconi, Riccardo Cicchi

¹University of Florence, Department of Evolutionary Biology Leo Pardi, Florence, Italy.

Journal of Biomedical Optics

|June 28, 2012

Summary

Second-harmonic generation (SHG) microscopy images unstained living tissues by analyzing light interactions. This technique reveals molecular organization and conformational changes in vivo, advancing molecular diagnostics.

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Area of Science:

Biomedical Optics
Nonlinear Microscopy
Molecular Imaging

Background:

Second-harmonic generation (SHG) microscopy is a key technique for imaging unstained living tissues.
It probes molecular and supramolecular organization by analyzing light interactions.
SHG's sensitivity to emitter distribution is based on coherent summation of the second-harmonic response.

Purpose of the Study:

To review the physical basis of SHG microscopy.
To examine experimental applications for probing molecular organization and its alterations in biomedically relevant conditions.
To describe SHG polarization anisotropy for studying molecular conformation in vivo.

Main Methods:

Review of the physical principles of SHG.
Analysis of polarized SHG for 3D emitter distribution.
Application of SHG polarization anisotropy for myosin conformation studies.

Main Results:

SHG microscopy provides micrometer-scale resolution in deep tissues.
Polarized SHG is sensitive to the 3D distribution of molecules.
SHG polarization anisotropy reveals molecular conformation, demonstrated in muscle myosin studies.

Conclusions:

SHG microscopy offers a powerful platform for in vivo molecular diagnostics.
It combines nonlinear microscopy advantages with molecular structure probing.
This technology significantly advances biomedical optics and in vivo diagnostics.