Document Detail

Hyperspectral imaging: a novel approach for microscopic analysis.
MedLine Citation:
PMID:  11260591     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: The usefulness of the light microscope has been dramatically enhanced by recent developments in hardware and software. However, current technologies lack the ability to capture and analyze a high-resolution image representing a broad diversity of spectral signatures in a single-pass view. We show that hyperspectral imaging offers such a technology. METHODS AND RESULTS We developed a prototype hyperspectral imaging microscope capable of collecting the complete emission spectrum from a microscope slide. A standard epifluorescence microscope was optically coupled to an imaging spectrograph, with output recorded by a CCD camera. Software was developed for image acquisition and computer display of resultant X--Y images with spectral information. Individual images were captured representing Y-wavelength planes, with the stage successively moved in the X direction, allowing an image cube to be constructed from the compilation of generated scan files. This prototype instrument was tested with samples relevant to cytogenetic, histologic, cell fusion, microarray scanning, and materials science applications.
CONCLUSIONS: Hyperspectral imaging microscopy permits the capture and identification of different spectral signatures present in an optical field during a single-pass evaluation, including molecules with overlapping but distinct emission spectra. This instrument can reduce dependence on custom optical filters and, in future imaging applications, should facilitate the use of new fluorophores or the simultaneous use of similar fluorophores.
R A Schultz; T Nielsen; J R Zavaleta; R Ruch; R Wyatt; H R Garner
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry     Volume:  43     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  2001 Apr 
Date Detail:
Created Date:  2001-03-22     Completed Date:  2001-05-21     Revised Date:  2013-05-20    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  239-47     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Wiley-Liss, Inc.
McDermott Center for Human Growth and Development, The University of Texas Southwestern Medical Center, Dallas, Texas 75235-8591, USA.
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MeSH Terms
Image Cytometry / instrumentation*,  methods
Image Processing, Computer-Assisted
Imaging, Three-Dimensional / instrumentation*,  methods
In Situ Hybridization, Fluorescence
Microscopy, Fluorescence / instrumentation*,  methods
Spectrometry, Fluorescence / instrumentation*,  methods
Staining and Labeling

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