Interleaved dual-wavelength multiphoton imaging system for heterologous FRET and versatile fluorescent protein excitation

M. Anna Zal; Michelle Nelson; Tomasz Zal

doi:10.1117/12.699242

16 February 2007 Interleaved dual-wavelength multiphoton imaging system for heterologous FRET and versatile fluorescent protein excitation

M. Anna Zal, Michelle Nelson, Tomasz Zal

Author Affiliations +

Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 64421Q (2007) https://doi.org/10.1117/12.699242
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

Broad two-photon cross sections of fluorescent proteins allow excitation with a single wavelength of a tunable femtosecond pulsed laser but the brightness is sub-optimal and the cross-talk prevents sensitized emission FRET imaging in heterologous systems. We present a novel arrangement of a resonant scanning microscope capable of fast interline dual wavelength femtosecond excitation of pairs of fluorophores. This allows optimal and selective excitation of mCerulean and mCitrine as well as FRET imaging using the principles of sensitized emission 3-cube imaging. Performance of the system in thin and thick specimens is discussed.

Citation Download Citation

M. Anna Zal, Michelle Nelson, and Tomasz Zal "Interleaved dual-wavelength multiphoton imaging system for heterologous FRET and versatile fluorescent protein excitation", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64421Q (16 February 2007); https://doi.org/10.1117/12.699242

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