Quantitative Colocalization Analysis of Multicolor Confocal Immunofluorescence Microscopy Images: Pushing Pixels to Explore Biological Phenomena
Open Access
- 1 January 2007
- journal article
- Published by Japan Society of Histochemistry & Cytochemistry in ACTA HISTOCHEMICA ET CYTOCHEMICA
- Vol. 40 (4), 101-111
- https://doi.org/10.1267/ahc.07002
Abstract
Quantitative colocalization analysis is an advanced digital imaging tool to examine antigens of interest in immunofluorescence images obtained using confocal microscopes. It employs specialized algorithms to estimate the degree of overlap of fluorescence signals and thus enables acquiring important new information not otherwise obtainable using qualitative approaches alone. As raw confocal images have high levels of background, they should be prepared to become suitable for reliable calculation of colocalization coefficients by correcting it. We provide concise theoretical basis of quantitative colocalization analysis, discuss its limitations, and describe proper use of the technique. The use of quantitative colocalization analysis is demonstrated by studying bile salt export pump and multidrug resistance associated protein 2 in the liver and major basic protein and platelet activating factor receptor antigens in conjunctiva. The review is focused on the applicability and correct interpretation of the results of colocalization coefficients calculations.Keywords
This publication has 33 references indexed in Scilit:
- Role of 12-lipoxygenase in volatile anesthetic-induced delayed preconditioning in miceAmerican Journal of Physiology-Heart and Circulatory Physiology, 2006
- Protection of adult rat cardiac myocytes from ischemic cell death: role of caveolar microdomains and δ-opioid receptorsAmerican Journal of Physiology-Heart and Circulatory Physiology, 2006
- Focal Adhesions in (Myo)fibroblasts Scaffold Adenylyl Cyclase with Phosphorylated CaveolinOnline Journal of Public Health Informatics, 2006
- Atrial natriuretic peptide promotes cardiomyocyte survival by cGMP-dependent nuclear accumulation of zyxin and AktJCI Insight, 2005
- Automated image analysis methods for 3-D quantification of the neurovascular unit from multichannel confocal microscope imagesCytometry Part A, 2005
- Experimental LPS-induced cholestasis alters subcellular distribution and affects colocalization of Mrp2 and Bsep proteins: A quantitative colocalization studyMicroscopy Research and Technique, 2005
- Colocalization analysis yields superior results after image restorationMicroscopy Research and Technique, 2004
- Endotoxin stimulates hydrogen peroxide detoxifying activity in rat hepatic endothelial cellsHepatology, 1996
- Endotoxin stimulates hydrogen peroxide detoxifying activity in rat hepatic endothelial cellsHepatology, 1996
- The point‐spread function of a confocal microscope: its measurement and use in deconvolution of 3‐D dataJournal of Microscopy, 1991