Effects of using a metal layer in total internal reflection fluorescence microscopy |
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| Authors: | W.T. Tang E. Chung Y. Kim P.T.C. So C.J.R. Sheppard |
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| Affiliation: | (1) Computation and Systems Biology, Singapore-MIT Alliance, National University of Singapore, 117576 Singapore, Singapore;(2) Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;(3) Department of Mechanical Engineering and Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;(4) Division of Bioengineering and Department of Diagnostic Radiology, National University of Singapore, 117576 Singapore, Singapore |
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| Abstract: | This proceeding examines the characteristics of imaging through a metal-coated glass cover slip using total internal reflection fluorescence microscopy (TIRFM). Through back and front focal plane imaging of sub-diffraction-limited fluorescent beads, the experimental characteristics of the emission are compared with theoretical simulations. Furthermore, at the angle at which surface plasmon resonance occurs, we show that the anisotropic emission of the fluorescent beads collected through the metal layer results in a irregular point spread function that has a donut-like structure with multiple concentric rings. PACS 42.30.Va; 73.20.Mf |
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