Open AccessUltrafast third harmonic micro-spectroscopy reveals a two-photon resonance in human hemoglobin
Author(s) -
Omar Clay,
Chris B. Schaffer,
Jeffrey A. Squier,
David Kleinfeld
Publication year - 2006
Publication title -
proceedings of spie, the international society for optical engineering/proceedings of spie
Language(s) - English
Resource type - Conference proceedings
SCImago Journal Rank - 0.192
H-Index - 176
eISSN - 1996-756X
pISSN - 0277-786X
DOI - 10.1117/12.659110
Subject(s) - ultrashort pulse , spectroscopy , materials science , laser , optics , high harmonic generation , nonlinear optics , microscopy , second harmonic generation , photon , two photon excitation microscopy , hemoglobin , resonance (particle physics) , optoelectronics , nuclear magnetic resonance , fluorescence , atomic physics , physics , chemistry , biochemistry , quantum mechanics
The recently developed technique of ultrafast third harmonic generation (THG) micro-spectroscopy is discussed. The approach is easily adapted to a standard laser scanning microscope and allows for two and three photon resonances to be identified in non-fluorescent unlabeled samples. This work provides nonlinear microscopists with a tool for further understanding the contrast and damage mechanisms they will encounter under nonlinear excitation. Here, we use THG micro-spectroscopy to investigate the nonlinear optical properties of hemoglobin over the spectral range of 770 -1000 nm with 100-fs duration, ~1-nJ energy laser pulses. We demonstrate the ability of this approach to distinguish different ligand binding states in physiological solutions of human hemoglobin.
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