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Published August 2005 | Published
Journal Article Open

Single-Biomolecule Resolution Imaging with an Optical Microscope

Abstract

A Fluorescence Apertureless Near-field Scanning Optical Microscope (FANSOM) has been developed with FWHM optical resolution below 10 nm when imaging at ~600 nm wavelengths [1]. The apparatus combines an epi fluorescence optical microscope and an atomic force microscope (AFM) to obtain single-molecule sensitivity and optical resolution limited by the sharpness of the AFM probe. The AFM probe is used to stimulate or reduce the detected fluorescence emission rate depending on the AFM tip material and the polarization of the excitation light. The probe-sample interaction is described by near-field dipole-dipole physics, resulting in a stimulated emission rate that varies by r^6. When tapping the probe over the substrate being imaged, the near-field component is sharply modulated at that tapping frequency, thereby enabling separation from the far-field background during post-processing. Images of fluorescent single-molecules taken in a physiological environment will be presented.

Additional Information

© 2005 Microscopy Society of America. Published online: 01 August 2005. Extended abstract of a paper presented at Microscopy and Microanalysis 2005 in Honolulu, Hawaii, USA, July 31-August 4, 2005. This work was supported by NASA/JPL Research and Technology Development and Bio-Nano Programs, the Caltech President's Fund, NIH, Pharmagenomics and Arrowhead Research.

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