28 February 2006 Ultrafast third harmonic micro-spectroscopy reveals a two-photon resonance in human hemoglobin
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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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Omar Clay, Chris B. Schaffer, Jeffrey A. Squier, 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); doi: 10.1117/12.659110; https://doi.org/10.1117/12.659110

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