Colocalization Colormap

Independent fluorescent labeling of two different components is a commonly used  method  to determine their co-localization.  This website  contains  an  ImageJ  plugin  for automatic quantification  and  visualization of co-localized fluorescent signals. The plugin is based on the method originally described by Jaskolski et. al. (2005). This algorithm computes correlation of  intensities between pairs  of  individual  pixels in  two  different  channels. Results   are  presented  as  a  co-localization  color map  where  hot colors represent positive correlation (co-localization), whereas colors that are cold represent negative correlation (exclusion).

Spatial representation of colocalization        

Correlation between pair of pixels (described by x,y coordinates in the image)  is calculated  according  to the formula below:
 
 

Distribution  of  nMDP  (normalized   mean  deviation  product) values  (ranging from -1 to 1)  is  visualized  with a color  scale. Negative indexes are  represented  by  cold  colors (exclusion). Indexes above 0 are represented by hot colors (co-localization).








Automatic  co-localization  calculation using Colocalization Colormap plugin.
(A) Neuronal  body(asterisks)  and  dendritic  processes  (arrows)
stained with antibody against MAP2 protein.
(B) Immunoreactivity  of  CD44  adhesion  molecule  in  the brain.
(C-D) Color map  presenting  co-localization   between   adhesion molecule and MAP2 neuronal marker.