Document Detail

Full spectrum filterless fluorescence microscopy.
MedLine Citation:
PMID:  20055924     Owner:  NLM     Status:  MEDLINE    
In conventional microscopes, fluorescence emission is separated from the backscattered illumination using the Stokes shift, whereby the emission occurs at a longer wavelength to the excitation. Such separation is usually achieved through a combination of wavelength filters that divide the spectrum into mutually exclusive excitation and emission bands. It is therefore impossible in these microscopes to access the full excitation/emission spectrum of the specimen in a single image. We report on a microscope that acquired fluorescence images using illumination across the spectral range 450-680 nm; the full emission spectrum was detected simultaneously across the same range. The microscope was also combined with structured illumination optical sectioning to give three-dimensionally resolved images with improved background rejection. Full spectrum fluorescence images of biological specimens are demonstrated. As this system is more versatile than the standard fluorescence microscope, it could be of benefit in many fluorescence imaging applications.
M J Booth; A Jesacher; R Juskaitis; T Wilson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microscopy     Volume:  237     ISSN:  1365-2818     ISO Abbreviation:  J Microsc     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-08     Completed Date:  2010-03-09     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  England    
Other Details:
Languages:  eng     Pagination:  103-9     Citation Subset:  IM    
Department of Engineering Science, University of Oxford Parks Road, Oxford OX13PJ, United Kingdom.
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MeSH Terms
Cell Line
Fluorescent Dyes
Imaging, Three-Dimensional
Microscopy, Fluorescence / instrumentation*
Sensitivity and Specificity
Grant Support
BB/E01240X/1//Biotechnology and Biological Sciences Research Council
Reg. No./Substance:
0/Fluorescent Dyes

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