| The effects of spherical aberration on multiphoton fluorescence excitation microscopy. | |
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MedLine Citation:
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PMID: 21118240 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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Multiphoton fluorescence excitation microscopy is almost invariably conducted with samples whose refractive index differ from that of the objective immersion medium, conditions that cause spherical aberration. Due to the quadratic nature of multiphoton fluorescence excitation, spherical aberration is expected to profoundly affect the depth dependence of fluorescence excitation. In order to determine the effect of refractive index mismatch in multiphoton fluorescence excitation microscopy, we measured signal attenuation, photobleaching rates and resolution degradation with depth in homogeneous samples with minimal light scattering and absorption over a range of refractive indices. These studies demonstrate that signal levels and resolution both rapidly decline with depth into refractive index mismatched samples. Analyses of photobleaching rates indicate that the preponderance of signal attenuation with depth results from decreased rates of fluorescence excitation, even in a system with a descanned emission collection pathway. Similar results were obtained in analyses of fluorescence microspheres embedded in rat kidney tissue, demonstrating that spherical aberration is an important limiting factor in multiphoton fluorescence excitation microscopy of biological samples. |
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Authors:
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P A Young; S G Clendenon; J M Byars; R S Decca; K W Dunn |
Publication Detail:
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Type: Journal Article Date: 2010-10-11 |
Journal Detail:
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Title: Journal of microscopy Volume: 242 ISSN: 1365-2818 ISO Abbreviation: J Microsc Publication Date: 2011 May |
Date Detail:
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Created Date: 2011-04-04 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0204522 Medline TA: J Microsc Country: England |
Other Details:
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Languages: eng Pagination: 157-65 Citation Subset: IM |
Copyright Information:
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© 2010 The Authors Journal of Microscopy © 2010 Royal Microscopical Society. |
Affiliation:
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Department of Medicine, Division of Nephrology, Indiana University School of Medicine, Indianapolis, Indiana, U.S.A. Department of Physics, Indiana University, Bloomington, Indiana, U.S.A. Department of Physics, Indiana University-Purdue University Indianapolis, Indianapolis, Indiana, U.S.A. |
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