Enhancing the axial resolution of fluorescence microscopy using two-photon confocal configuration

Authors

Serafin Delica National Institute of Physics, University of the Philippines Diliman
Carlo Mar Blanca National Institute of Physics, University of the Philippines Diliman
Bernardino Buenaobra National Institute of Physics, University of the Philippines Diliman
Caesar Saloma National Institute of Physics, University of the Philippines Diliman

Abstract

We increase the axial resolution of a fluorescence microscope by utilizing two-photon excitation in a confocal configuration. This resolution increase is rooted on the nonlinear nature of the excitation process which was verified to have a power dependence of 2.02 – the exact excitation signature of a two-photon absorption process and on the detection discrimination in provided by the confocal pinhole. The two-photon confocal arrangement improves the resolution to 1.6 μm, 3 times sharper than two-photon widefield configuration and 96 times sharper than the single photon widefield case. The microscope is used to image 10 μm fluorescent spheres with minimal photobleaching.

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Issue

2004: Proceedings of the 22nd Samahang Pisika ng Pilipinas Physics Congress

Article ID

SPP-2004-PA-18

Section

Poster Session PA

Published

2004-10-25

How to Cite

[1]

S Delica, CM Blanca, B Buenaobra, and C Saloma, Enhancing the axial resolution of fluorescence microscopy using two-photon confocal configuration, Proceedings of the Samahang Pisika ng Pilipinas 22, SPP-2004-PA-18 (2004). URL: https://proceedings.spp-online.org/article/view/SPP-2004-PA-18.