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Colocalization Analysis in Fluorescence Microscopy
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Cancer and Vascular Biology. (Claesson-Welsh)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Cell Biology.
2012 (English)In: Cell Imaging Techniques: Methods and Protocols / [ed] Taatjes, Douglas J. & Roth, Jürgen, New York: Humana Press, 2012, 97-109 p.Chapter in book (Refereed)
Abstract [en]

The measurement of colocalization requires images of two fluorophores that are aligned, with no cross talk, and that the intensities remain within the response range of the microscope. Quantitation depends upon differentiating between the presence and absence of fluorescence, and measurements should be made within biologically relevant regions of interest. Co-occurrence can be measured simply by area or with the M1 and M2 coefficients, and should be compared to random distributions. Correlation analysis should use the Pearson and Spearman coefficients, which need to be measured by replicate based noise corrected correlation to eliminate errors arising from differences in image quality. Ideally, both co-occurrence and correlation should be reported.

Place, publisher, year, edition, pages
New York: Humana Press, 2012. 97-109 p.
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 319
Keyword [en]
colocalization, co-occurrence, correlation, pearson correlation coefficient, RBNCC
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-183653DOI: 10.1007/978-1-62703-056-4_5ISBN: 978-1-62703-055-7 (print)OAI: oai:DiVA.org:uu-183653DiVA: diva2:563664
Available from: 2012-10-31 Created: 2012-10-31 Last updated: 2012-11-01Bibliographically approved

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