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

Authors

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