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PERSPECTIVES IN CELL PHYSIOLOGY

Image acquisition for colocalization using optical microscopy

¹Department of Cellular and Physiological Sciences, University of British Columbia, Life Sciences Institute, Vancouver, British Columbia, Canada; and ²Department of Physiology, University of Arizona, Arizona Health Sciences Center, Tucson, Arizona

Submitted 29 February 2008 ; accepted in final form 17 March 2008

ABSTRACT

Colocalization, in which images of two or more fluorescent markers are overlaid, and coincidence between the probes is measured or displayed, is a common analytical tool in cell biology. Interpreting the images and the meaning of this identified coincidence is difficult in the absence of basic information about the acquisition parameters. In this commentary, we highlight important factors in the acquisition of images used to demonstrate colocalization, and we discuss the minimum information that authors should include in a manuscript so that a reader can interpret both the fluorescent images and any observed colocalization.