IMAGEJ COLOCALIZATION HOW TO

But if there is nothing to compare them to, what do they mean? A statistical significance test was derived by Costses to evaluate the probability that the measured value of Pearson's correlation, r between the two colour channels is significantly greater than values of r that would be calculated if there was only random overlap of the same information. These coefficients measure the amount or degree of colocalization, or rather correlation and co-occurrence respectively (but should not be expressed as % values, because that is not how they are defined). Values range from 0 to 1, expressing the fraction of intensity in a channel that is located in pixels where there is above zero (or threshold) intensity in the other colour channel. You can get more details in Manders et al.

Proportional to the amount of fluorescence of the colocalizing pixels or voxels in each colour channel. Noise makes the value closer to 0 than it should be. The result is +1 for perfect correlation, 0 for no correlation, and -1 for perfect anti-correlation. It is not sensitive to differences in mean signal intensities or range, or a zero offset between the two components. Here just are two of many colocalization coefficients to express the intensity correlation of colocalizing objects in each component of a dual-color image: Methods of colocalization analysis Pixel intensity spatial correlation analysis

IMAGEJ COLOCALIZATION HOW TO

For one place to start reading about colocalisation and for how to correctly capture quantitative fluorescence microscopy images suitable for colocalisation analysis, look here: Image Processing Courses at BioDIP, Dresden.

First you have to define what you mean by colocalisation, and that is not trivial. Suppose you are given some images by a colleague, or have some images of your own, and you want to measure the amount of colocalisation between two of the dyes or stains in the images.

5.1 Check image data for problems and suitability for analysis.

4 ImageJ plugins for colocalization analysis.

2 Why scatter plots instead of colour merge images?.

1.1.3 Do state the spatial resolution explicitly!.1.1.1 Pixel intensity spatial correlation analysis.

Imagej colocalization