Modern biological research particularly in the fields of developmental and cell biology has been transformed by the rapid evolution of the light microscope. The light microscope, long a mainstay of the experimental biologist, is now used for a wide array of biological experimental scenarios and sample types. Much of the great developments in advanced biological imaging have been driven by the digital imaging revolution with powerful processors and algorithms. In particular, this combination of advanced imaging and computational analysis has resulted in the drive of the modern biologist to not only visually inspect dynamic phenomena, but to quantify the involved processes. This need to quantitate images has become a major thrust within the bioimaging community and requires extensible and accessible image processing routines with corresponding intuitive software packages. Novel algorithms both made specifically for light microscopy or adapted from other fields, such as astronomy, are available to biologists, but often in a form that is inaccessible for a number of reasons ranging from data input issues, usability and training concerns, and accessibility and output limitations. The biological community has responded to this need by developing open source software packages that are freely available and provide access to image processing routines. One of the most prominent is the open-source image package ImageJ. In this review, we give an overview of prominent imaging processing approaches in ImageJ that we think are of particular interest for biological imaging and that illustrate the functionality of ImageJ and other open source image analysis software. WIREs Dev Biol 2017, 6:e260. doi: 10.1002/wdev.260 For further resources related to this article, please visit the WIREs website.
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