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In this episode, Erin and Beth dive into the Threshold module in CellProfiler and explain how to pick the right thresholding algorithm so that you can best distinguish stuff from not stuff. They explain some concepts in thresholding with “No math. Concepts of math, but no math.” As if there as any question about how geeky they are, Erin and Beth both declare their favorite thresholding algorithms (Otsu 3-class, baby!). This is the second episode in a 3-part series on thresholding in CellProfiler. The first episode is available at • Thresholding in CellProfiler Part 1: Adapt... . The ~*~Sparkle~*~ episode is at • Identifying Speckles in CellProfiler - Ask... . #science #openscience #microscopy #tutorial #opensource #biology #imageanalysis #segmentation #cellprofiler ================================================== On Ask Erin/Dear Beth, bioimage analysis experts Beth Cimini and Erin Weisbart, of the Imaging Platform at the Broad Institute of MIT and Harvard, answer your image analysis questions! Whether it’s about tuning or troubleshooting, segmenting or measuring, Beth and Erin have opinions and by golly are they going to share them! Using mostly CellProfiler, but with expertise in many other image analysis softwares such as ImageJ/FIJI, Cellpose, Piximi, and more, no image analysis question is too hard, so ask away! Submit your own questions at broad.io/AskErinDearBeth Images and pipelines we’ve used are available for you to download and play with at https://ciminilab.github.io/AskErin_D...