Probing live samples in second-harmonic generation microscopy using specific markers and fluorescent proteins

E. De Meulenaere; R. Paesen; S. Psilodimitrakopoulos; M. Ameloot; P. Loza-Alvarez; J. Vanderleyden; K. Clays

doi:10.1117/12.907498

9 February 2012 Probing live samples in second-harmonic generation microscopy using specific markers and fluorescent proteins

E. De Meulenaere, R. Paesen, S. Psilodimitrakopoulos, M. Ameloot, P. Loza-Alvarez, J. Vanderleyden, K. Clays

Author Affiliations +

Proceedings Volume 8226, Multiphoton Microscopy in the Biomedical Sciences XII; 82263C (2012) https://doi.org/10.1117/12.907498
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

In an effort to complement cellular two-photon excited fluorescence (TPEF) microscopy with structural information from second-harmonic generation (SHG) imaging, we investigated the applicability of fluorescent proteins for SHG imaging. In the first stage, the first hyperpolarizability β, a measure for the second-order nonlinear optical properties of a molecule, was determined for several fluorescent proteins. In a second stage, an established HeLa cell line expressing a membrane protein labeled with a fluorescent protein, was adapted and imaged using simultaneous TPEF and SHG microscopy. The contour of stretched cells observed in these experiments was proven to be originating in microtubules instead of the fluorescent proteins.

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E. De Meulenaere, R. Paesen, S. Psilodimitrakopoulos, M. Ameloot, P. Loza-Alvarez, J. Vanderleyden, and K. Clays "Probing live samples in second-harmonic generation microscopy using specific markers and fluorescent proteins", Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 82263C (9 February 2012); https://doi.org/10.1117/12.907498

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