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CoLocalizer Pro
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CoLocalizer Pro is a scientific software application, developed by CoLocalization Research Software, that allows researchers to analyze colocalization in the images obtained using fluorescence microscopy. Due to high popularity of Macintosh computers in medicine and biology, it is designed specifically for Mac OS X operating system. To bring the full advantage of Mac OS X, the software is written exclusively in Cocoa. CoLocalizer Pro is used in universities and research institutions worldwide. Lite version of the software, CoLocalizer Express, is geared toward students and beginners. The use of CoLocalizer Pro software is cited by various top impact factor journals, including Cell, Nature Neuroscience, Nature Methods, Journal of Clinical Investigation, Journal of Cell Biology, PNAS, etc. In October 2011 issue, renown Nature Protocols published a CoLocalizer Pro-based protocol to quantify spatial correlations of fluorescent markers. The protocol described in details the steps needed to be taken to perform quantification of colocalization of synergetic proteins. Overview CoLocalizer Pro employs a pixel-based method to estimate the degree of colocalization by calculating specialized coefficients and prepares obtained results for presentation and publication. The following coefficients can be calculated: , overlap coefficient (R), overlap coefficients k<sub>1</sub>-k<sub>2</sub>, colocalization coefficients m<sub>1</sub>-m<sub>2</sub>, and colocalization coefficients M<sub>1</sub>-M<sub>2</sub>. The workflow is designed to ensure that the results are accurate and reproducible. To provide the accuracy of calculations, the image background noise is corrected prior to the analysis and therefore does not impact the results. Another notable feature of the software is the option to save the results of coefficients calculations and image analysis as session reports with all the data combined in a single file for compact storage and further reference. The information about the degree of colocalization of proteins can play a role in understanding their functional significance with various medical and biological implications. Features * Views fluorescence microscopy images with colocalization at different magnifications and angles, merges single channel images into one or splits merged ones according to a pair of channels * Analyzes image stacks * Saves versions of analyzed images * Directly imports images from microscopes of analysis * Corrects background in the images to ensure reliability of quantification * Quantifies colocalization by calculating specialized coefficients * Examines image ROI, scattergram ROI, reveals colocalized or selected pixels, traces pixels of a particular color * Saves calculations data in various file formats, such as Microsoft Excel, Text, PDF, and HTML * Exports results as colocalization reports with all the coefficients and image data stored conveniently in one place Free colocalization benchmark sets Quantitative data obtained using CoLocalizer Pro can be validated with the help of free downloadable image sets from the Colocalization Benchmark Source. By calculating and comparing the values of coefficients on their images versus benchmark images from CBS, researchers can interpret their calculations. 
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