Ultrafast third harmonic micro-spectroscopy reveals a two-photon resonance in human hemoglobin

Omar Clay; Chris B. Schaffer; Jeffrey A. Squier; David Kleinfeld

doi:10.1117/12.659110

28 February 2006 Ultrafast third harmonic micro-spectroscopy reveals a two-photon resonance in human hemoglobin

Omar Clay, Chris B. Schaffer, Jeffrey A. Squier, David Kleinfeld

Proceedings Volume 6108, Commercial and Biomedical Applications of Ultrafast Lasers VI; 610809 (2006) https://doi.org/10.1117/12.659110
Event: Lasers and Applications in Science and Engineering, 2006, San Jose, California, United States

Abstract

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.

Citation Download Citation

Omar Clay, Chris B. Schaffer, Jeffrey A. Squier, and David Kleinfeld "Ultrafast third harmonic micro-spectroscopy reveals a two-photon resonance in human hemoglobin", Proc. SPIE 6108, Commercial and Biomedical Applications of Ultrafast Lasers VI, 610809 (28 February 2006); https://doi.org/10.1117/12.659110

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