Journal
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
Volume 60, Issue 6, Pages 428-438Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1369/0022155412442897
Keywords
human muscle; myosin heavy chain; myotubes; myotube size; myotube area; myotube diameter; semiquantitative image analysis; layers; fluorescence intensity
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Funding
- King's College London
- Department of Health via the National Institute for Health Research (NIHR) comprehensive Biomedical Research Centre
- St Thomas' NHS Foundation Trust
- King's College Hospital NHS Foundation Trust
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The accurate measurement of the morphological characteristics of cells with nonuniform conformations presents difficulties. We report here a straightforward method using immunofluorescent staining and the commercially available imaging program Adobe Photoshop, which allows objective and precise information to be gathered on irregularly shaped cells. We have applied this measurement technique to the analysis of human muscle cells and their immunologically marked intracellular constituents, as these cells are prone to adopting a highly branched phenotype in culture. Use of this method can be used to overcome many of the long-standing limitations of conventional approaches for quantifying muscle cell size in vitro. In addition, wider applications of Photoshop as a quantitative and semiquantitative tool in immunocytochemistry are explored. (J Histochem Cytochem 60:428-438, 2012)
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