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Help! Interpreting image-based profiles

Fernanda Garcia-Fossa & Anne Carpenter In a typical quantitative microscopy experiment, biologists choose fluorescent biomarkers and measure particular features (that is “metrics”) that they hypothesize will be perturbed in their samples. But in image-based profiling, you aim to let the cells tell...

Customizing a Model for Fiber Segmentation, Part 4: Analyzing the Results

Melissa Gillis In the final section of this blogpost series, I will discuss some of the benefits and drawbacks of this model and discuss some of the data I acquired from the images. This model was able to successfully segment the images that were not able to be accurately segmented by Pearl’s...

Customizing a Model for Fiber Segmentation, Part 2: Creating an ilastik Model

Melissa Gillis In the first part of this blogpost I described some of the tools I tried to accurately segment collagen IV fibers including, CellProfiler, Cellpose , and Piximi . Unfortunately none of these methods were successful in accurately segmenting the fibers so I decided to develop a machine...

Help! How does the Robust Background method work?

Pearl V. Ryder The Robust Background algorithm is a powerful algorithm for automatically setting thresholds to segment objects of interest when your image contains mostly background. However, since it contains the largest number of tunable parameters of any thresholding algorithm in CellProfiler, it...

Help! Why does CellProfiler say it can’t find any valid image sets?

Beth Cimini Defining the input to CellProfiler can be the hardest part of getting your pipeline set up and your analysis underway. Incoming images are configured in the first 4 modules of CellProfiler – Images, Metadata, NamesAndTypes, and Groups – which offer lots of flexibility. But it’s sometimes...

Help! What are these three different modules to identify objects?

Beth Cimini It can be confusing when you’re trying to set up your first pipeline to figure out which modules to use to generate your objects! A helpful way to understand the difference between Identifying Primary, Secondary, and Tertiary objects: Primary objects are segmented independently from any...