Workflow

OligoMacro Toolset, is an ImageJ macro-toolset aimed at isolating oligodendrocytes from wide-field images, tracking isolated cells, characterizing processes morphology along time, outputting numerical data and plotting them. It takes benefit of ImageJ built-in functions to process images and extract data, and relies on the R software in order to generate graphs.

The research goal of this paper was to provide unbiased counts of labeled astrocytes and to estimate the area they cover, further to develop tools for defining the orientation of coupling within astrocyte networks under different stimuli.

In order to count the astrocytes and estimate the area they cover the following steps were used in this software.

Pre-processing: z-project (using max intensity); split channels; subtract background; remove outliers.

Segmentation: adjust threshold and convert to a binary file; Watershed.

Cell counting: Analyze particles

Measure Astrocytic network area: select a ROI using the polygon tool; set measurements (area); ROI manager -> add the traced polygon; measure.