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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...

Rebecca Senft With slide scanners and other automated, high-throughput microscopes becoming more and more common, it’s important to understand how to work with the large image files they produce. Whole slide file formats (e.g., .mrxs, .svslide, .svs, .vms) are often massive when uncompressed (>40 GB...

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...

Mario Costa Cruz If you have color images stained with light-absorbing dyes, the UnmixColors module might be able to separate the dyes into separate channels so that they can be analyzed separately. Separating the dyes into channels can be a difficult task with traditional methods such as split...

Barbara Diaz-Rohrer If you are working with single-channel images, you can just drag a few images into CellProfiler and start making your pipeline. Most of us, however, have images with multiple channels or more complex image metadata that you need to tell CellProfiler how to identify.The tutorial...

Melissa Gillis Segmenting fibers using traditional cell segmentation software can be challenging. Because most biological cell segmentation tools were designed for cells, most programs have difficulty identifying and distinguishing longer and thinner objects such as fibers. Also, unlike cells...

Melissa Gillis In the previous two sections of this blogpost I explained how I analyzed multiple methods to segment fluorescent images of collagen IV fibers, and generated accurate segmentation using an ilastik machine learning model. In this section I will explain how I constructed a CellProfiler...

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...

Nasim Jamali As creators of open source tools, we spend a lot of time thinking about user needs - what kinds of images people are working with these days, what tools are working well, where the pain points are, and how easy it is to learn how to do any of this in the first place. While things like...

Rebecca Senft Workflow update June 2024 update Our recommended normalization workflow is evolving. Please see https://github.com/broadinstitute/jump-profiling-recipe/ for our latest developments on the normalization strategy in specific and aprofiling workflow in general. How to Normalize Cell...