Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning

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We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies. (C) 2013 Optical Society of America
Publisher
OPTICAL SOC AMER
Issue Date
2013-07
Language
English
Article Type
Article
Citation

OPTICS EXPRESS, v.21, no.15, pp.17839 - 17848

ISSN
1094-4087
DOI
10.1364/OE.21.017839
URI
http://hdl.handle.net/10203/192487
Appears in Collection
ME-Journal Papers(저널논문)
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