A popular approach was introduced by Sylvain Costes, who utilized Pearson's correlation coefficient as a tool for setting the thresholds required by M1 and M2 in an objective fashion.
[6] In addition, due to the tendency of fluorescence images to contain a certain amount of out-of-focus signal, and poisson shot and other noise, they usually require pre-processing prior to quantification.
Up to now, most frequently used methods to quantify colocalization calculate the statistical correlation of pixel intensities in two distinct microscopy channels.
[13] Some impermeable fluorescent zinc dyes can detectably label the cytosol and nuclei of apoptizing and necrotizing cells among each of four different tissue types examined.
The molecules are detected as spots appearing on the surface in real-time, and their locations are found to within 10-20 nm by fitting of point-spread functions.
Since typical sizes of biomolecules are on the order of 10 nm, this precision is usually sufficient for calling of molecular interactions [17] For the purpose of better interpretation of the results of qualitative and quantitative colocalization studies, it was suggested to use a set of five linguistic variables tied to the values of colocalization coefficients, such as very weak, weak, moderate, strong, and very strong, for describing them.