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Anna Bosch: n-bits Colocalization Macro for Image J

Abstract

Colocalization analysis compares the spatial localization of two fluorescent molecules. In colocalization quantitative fluorescence 12 or 16 bits images depth are usually required to have a broader dynamic range. Here we have developed a macro instruction for Image J (Wayne Rasband, National Institute of Health, USA) based on the colocalization highlighter plugin (P. Bourdoncle, Institute Jacques Monod, Service Imagerie, Paris, France) to highlight colocalization pixels from n bits images. This macro generates an image of colocalized pixels (binary) through the analysis pixel to pixel from each channel. This image will be defined by two criteria: 1) pixels with intensities higher than user-defined thresholds of their respective channel. 2) pixels where the ratios of intensities of the two channels are above a user-defined ratio (0-1).

Features from this macro are:

  • It works with n bits images and stacks of the two channel images.
  • User can decide parameters of colocalization criteria during the process: threshold and ratio among the two channels.
  • The values of the colocalized points are converted automatically to the real value of each channel and normalized to the total of each channel (proportion of colocalization)

Keywords

colocalization, macro

Administrative data

Presenting author: Anna Bosch
Organisation: Universitat de Barcelona-IDIBAPS

co-authors: Pol Solans, Neus Abella and Maria Calvo

Type: Poster (portrait)

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