Abstract
Spectral self-interference fluorescence microscopy (SSFM) has been recently developed to determine the axial position of fluorescent emitters placed on reflecting dielectric structures. In this paper, we review SSFM with emphasis on its axial localization capabilities. We show that there is a tradeoff between the numerical aperture (NA) of the microscopy system and the axial localization fidelity. To use high-NA objectives for better lateral resolution, we describe and demonstrate a high-NA 4Pi microscopy system that performs spectral self-interference microscopy for axial localization of less than 2 nm. We demonstrate axial localization by using artificial samples. We also probe the membrane topography oí a Shigella flexneri bacterium, several micrometers away from a solid support.
Original language | English (US) |
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Pages (from-to) | 217-225 |
Number of pages | 9 |
Journal | IEEE Journal on Selected Topics in Quantum Electronics |
Volume | 14 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2008 |
Funding
Manuscript received September 21, 2007; revised October 19, 2007. This work was supported in part by the U.S. National Science Foundation under Grant DBI 0138425 and Award EEC-9987821 of the Engineering Research Centers Program, in part by the Air Force Office of Scientific Research under Grant MURI F-49620-03-1-0379, and in part by the National Institutes of Health, National Institute of Biomedical Imaging and Bio Engineering under Grant 5R01 EB00 756-03.
Keywords
- Fluorescence microscopy
- Localization
- Selfinterference
- Subcellular imaging
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Electrical and Electronic Engineering