The Colocalization Benchmark Source (CBS) is a free collection of downloadable images to test and validate the degree of colocalization of markers in fluorescence microscopy studies. It consists of computer-simulated images with exactly known (pre-defined) values of colocalization ranging from 0% to 90%. They can be downloaded as sets as well as separately.

Images were created using BioSim software as described previously. They simulate fluorescence microscopy images with a sampling rate of 4 pixels per resolution element. If the optical resolution is 200 nm, then the benchmark images are corresponding to images with a pixel size of 50 nm. All images are entirely background noise and out-of-focus fluorescence free. The size of images is 1024x1024 pixels.

These benchmark images can be used as reference points when quantifying colocalization in dual color fluorescence microscopy studies. The use of CBS will help to obtain more accurate and reliable information in quantitative colocalization experiments and, ultimately, improve understanding of the biological applicability of colocalization observations.

When you employed CBS images for reference in your publication(s), please let us know about it so that we can provide links to your paper(s) and results.

We are working on extending this database and its examples and we encourage fellow scientists to submit examples of their own images when CBS was used to validate colocalization. Please make sure to contribute to CBS by submitting your images.

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