Computational modeling of STED microscopy through multiple biological cells under one- and two-photon excitation

Andrew E. Mark; Mitchell A. Davis; Matthew S. Starosta; Andrew K. Dunn

doi:10.1117/12.2078345

9 March 2015 Computational modeling of STED microscopy through multiple biological cells under one- and two-photon excitation

Andrew E. Mark, Mitchell A. Davis, Matthew S. Starosta, Andrew K. Dunn

Author Affiliations +

Proceedings Volume 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII; 93300V (2015) https://doi.org/10.1117/12.2078345
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

While superresolution optical microscopy techniques afford enhanced resolution for biological applications, they have largely been used to study structures in isolated cells. We use the FDTD method to simulate the propagation of focused beams for STED microscopy through multiple biological cells. We model depletion beams that provide 2D and 3D confinement of the fluorescence spot and assess the effective PSF of the system as a function of focal depth. We compare the relative size of the STED effective PSF under one- and two-photon excitation. PSF calculations suggest that imaging is possible up to the maximum simulation depth if the fluorescence emission remains detectable.

Citation Download Citation

Andrew E. Mark, Mitchell A. Davis, Matthew S. Starosta, and Andrew K. Dunn "Computational modeling of STED microscopy through multiple biological cells under one- and two-photon excitation", Proc. SPIE 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII, 93300V (9 March 2015); https://doi.org/10.1117/12.2078345

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