An automated method to quantify and visualize colocalized fluorescent signals.
Journal of Neuroscience Methods. 2005-07-01; 146(1): 42-49
DOI: 10.1016/j.jneumeth.2005.01.012
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1. J Neurosci Methods. 2005 Jul 15;146(1):42-9.
An automated method to quantify and visualize colocalized fluorescent signals.
Jaskolski F(1), Mulle C, Manzoni OJ.
Author information:
(1)Laboratoire Physiologie Cellulaire de la Synapse, CNRS UMR 5091, Institut
François Magendie, Université Bordeaux 2, Rue C. Saint-Saëns, 33077 Bordeaux
Cedex, France.
The most commonly used method to analyze colocalization of fluorescent signal in
paired images is based on superimposition of images (« merging ») and visual
inspection. A method based on the comparison of the mean deviation of fluorescent
signal intensity has recently been proposed to quantify colocalization within a
user-defined area [Li Q, Lau A, Morris TJ, Guo L, Fordyce CB, Stanley EF. A
syntaxin 1, Galpha(o), and N-type calcium channel complex at a presynaptic nerve
terminal: analysis by quantitative immunocolocalization. J Neurosci
2004;24:4070-81]. Unfortunately, the latter quantification method does not
provide a spatial representation of the correlation between the two fluorescent
signals. Here we propose a new method that combines quantification and imaging of
colocalization. We describe an algorithm based on edge detection and calculation
of signal intensity deviation. The method is illustrated and validated on both
simulated images and experimental data. This new and automated method calculates
a correlation index (I(corr)) and generates an image of the correlated signals
from the two original images. In addition to help in comparing and quantifying
colocalization between two fluorescent stainings, this method can be adapted to
measure the distribution of ions, proteins, organelles and cells in a large array
of techniques.
DOI: 10.1016/j.jneumeth.2005.01.012
PMID: 15935219 [Indexed for MEDLINE]