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A double fluorescence staining protocol to determine the cross-sectional area of myofibers using image analysisA double fluorescence staining protocol was developed to facilitate computer based image analysis. Myofibers from experimentally treated (irradiated) and control growing turkey skeletal muscle were labeled with the anti-myosin antibody MF-20 and detected using fluorescein-5-isothiocyanate (FITC). Extracellular material was stained with concanavalin A (ConA)-Texas red. The cross-sectional area of the myofibers was determined by calculating the number of pixels (0.83 mu m(2)) overlying each myofiber after subtracting the ConA-Texas red image from the MF-20-FITC image for each region of interest. As expected, myofibers in the irradiated muscle were smaller (P < 0.05) than those in the non-irradiated muscle. This double fluorescence staining protocol combined with image analysis is accurate and less labor-intensive than classical procedures for determining the cross-sectional area of myofibers.
Document ID
20040173252
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Mozdziak, P. E.
(Muscle Biology Laboratory, University of Wisconsin and Department of Anatomy, Madison, 53706, United States)
Fassel, T. A.
Schultz, E.
Greaser, M. L.
Cassens, R. G.
Date Acquired
August 22, 2013
Publication Date
March 1, 1996
Publication Information
Publication: Biotechnic & histochemistry : official publication of the Biological Stain Commission
Volume: 71
Issue: 2
ISSN: 1052-0295
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Musculoskeletal

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