| Dark field Raman microscopy. | |
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MedLine Citation:
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PMID: 20568722 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Confocal Raman microscopy is often used for optical sectioning but is problematic when the sample plane of interest has a weak Raman cross-section/signal relative to areas that are out-of-focus. This is especially true for clinical samples in pathology, which consist of a thin tissue (approximately 5 microm) sample placed on a thick glass slide. Here, we recognize that the problem is the result of the extent of the illumination at the confocal plane being larger than the size of the sample and propose a dark field illumination scheme to efficiently reject substrate signals. The ability of several optical configurations in rejecting out-of-plane signal is investigated for two model systems: SU-8 photo resist over Teflon and SU-8 photo resist over polystyrene. The proposed reflective dark field approach, in which excitation converged to a focal point slightly above the focal plane of the collection optics, was found to be most effective in recording data from the sample. The proposed approach is validated by the rejection of substrate response (fluorescence) in spectra acquired from approximately 4 microm of breast tissue on a glass microscope slide. The proposed approach is easy to implement on existing confocal systems, has a straightforward optimization in acquiring data, and is not expected to result in loss of lateral resolution in mapping experiments. |
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Authors:
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Matthew V Schulmerich; Rohith Reddy; Anil K Kodali; Laura Jane Elgass; Krishnarao Tangella; Rohit Bhargava |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Analytical chemistry Volume: 82 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-07-29 Completed Date: 2010-11-04 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 6273-80 Citation Subset: IM |
Affiliation:
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Department of Bioengineering and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, 405 N. Mathews Ave. Urbana, Illinois 61801, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Epoxy Compounds
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chemistry Microscopy, Confocal / instrumentation*, methods Polymers / chemistry Polystyrenes / chemistry Polytetrafluoroethylene / chemistry Spectrum Analysis, Raman |
| Chemical | |
Reg. No./Substance:
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0/Epoxy Compounds; 0/Polymers; 0/Polystyrenes; 0/SU-8 compound; 9002-84-0/Polytetrafluoroethylene |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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