Greater signal and contrast in two-photon microscopy with ultrashort pulses

Lindsay R. Weisel; Peng Xi; Yair Andegeko; Vadim V. Lovozoy; Marcos Dantus

doi:10.1117/12.763549

15 February 2008 Greater signal and contrast in two-photon microscopy with ultrashort pulses

Lindsay R. Weisel, Peng Xi, Yair Andegeko, Vadim V. Lovozoy, Marcos Dantus

Proceedings Volume 6860, Multiphoton Microscopy in the Biomedical Sciences VIII; 68601O (2008) https://doi.org/10.1117/12.763549
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

Ultrashort <15 fs pulses are shown to provide higher fluorescence intensity, deeper sample penetration, and single laser selective excitation. To realize these advantages chromatic dispersion effects must be compensated. We use multiphoton intrapulse interference phase scan (MIIPS) to measure and then eliminate high-order distortions on pulses with a bandwidth greater than 100nm FWHM. Once compensated, the transform limited pulses deliver higher signal intensity, and this translates into deeper optical penetration depth with a high signal-to-noise ratio. By using a pulse shaper and taking advantage of the broad spectrum of the ultrafast laser, selective excitation of different cell organelles is observed due to the difference in nonlinear optical susceptibility of different chromophores without the use of an emission filter wheel.

Citation Download Citation

Lindsay R. Weisel, Peng Xi, Yair Andegeko, Vadim V. Lovozoy, and Marcos Dantus "Greater signal and contrast in two-photon microscopy with ultrashort pulses", Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 68601O (15 February 2008); https://doi.org/10.1117/12.763549

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