Human HT29 cells are fairly smooth and elliptical. This pipeline demonstrates how to accurately identify these cells and how to measurements cellular parameters such as morphology, count, intensity and texture.
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In contrast to the HT29 cells, Drosophila Kc167 cells are a highly textured and clumpy cell type. This pipeline demonstrates how to identify these clumpy cells and obtain morphological, intensity and texture measurements.
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A simple pipeline that identifies and counts tumors in a mouse lung, and then measures their size.
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This is a simple example of a DNA damage assay using single cell gel electrophoresis. Here, the measurement of interest is the length and intensity of the comet tail. Also, illumination correction is used to reduce background fluorescence prior to measurement. Also shown is a silver-stained comet example in which the percentage of DNA contained in the tail is calculated.
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CellProfiler is commonly used to count cells or other objects as well as percent-positives, by measuring the per-cell staining intensity. This pipeline shows how to do both of these tasks, and demonstrates how various modules may be used to accomplish the same result.
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This pipeline demonstrates how to classify and count objects on the basis of their measured features. The example identifies uniformly round objects, in this case, yeast colonies growing on a dish. The pipeline also shows how to load a template and align it to a cropped image, as well as how to use illumination correction to subtract for background illumination.
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This pipeline identifies patches of yeast growing in a 96 well plate, serving as an introduction to the grid defintion and identification modules.
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Tissue samples often have irregularly shaped cells with adjacent edges. This pipeline shows how to input a color tissue image, split it into its component channels, and then identify individual cells from a particular stain and record the number of neighbors that each cell has.
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In this example, cells are grown as a tissue monolayer. Rather than identifying individual cells, this pipeline quantifies the area occupied by the tissue sample.
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Illumination correction is often important for both accurate segmentation and for intensity measurements. This example shows how the CorrectIlluminationCalculate and CorrectIlluminationApply modules are used to compensate for the non-uniformities in illumination often present in microscopy images.
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Measuring the colocalization between fluorescently labeled molecules is a widely used approach to measure the degree of spatial coincidence and potential interactions among subcellular species (e.g., proteins). This example shows how the object identifcation and RelateObjects modules are used to measure the degree of overlap between two fluorescent channels.
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Imaging flow cytometry combines the high-throughput capabilities of conventional flow cytometry with single-cell imaging. CellProfiler can be used to analyze the resulting images from imaging flow cytometry, whether brightfield, darkfield, or fluorescence.
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These pipelines have been developed for high-throughput screens on C. elegans and extract measurements on a per-worm basis. The Worm Toolbox page has further details on this workflow, as well as video tutorials, pipelines and image data in addition to those described below. (from C Wählby et al. Methods, 2014) The BBBC also has C. elegans sample images and information, as well as assay "ground truth" of various kinds.
In this pipeline, we identify individual worms and extract shape and intensity measurements. Worm untangling requires a worm model, which is provided together with the pipeline. If adjusting the pipeline to fit your own data, worm detection will likely improve by creating a new worm model based on your own image data.
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Once worms are untangled, this pipeline shows how they can be straightened and aligned with a low-resolution worm atlas to extract localized intensity measurements and compare patterns of reporter signals. Included are steps for identifying secondary objects (fluorescent marker signals) and relating these objects to individual worms, enabling count of signals on a per-worm basis.
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This pipeline detects individual worms by worm untangling and finds sub-objects (fatty regions stained with oil red O) within the worms. Using bright-field data only, it detects fatty regions by intensity thresholding in a single image channel and relates the fatty regions to individual worms. This enables detection of rare phenotypes in heterogeneous populations, phenotypes that would be missed if population averages were observed. More data can be found on the BBBC.
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In this human cytoplasm-nucleus translocation assay, learn how to load a previously calculated illumination correction function for two separate channels, measure protein content in the nucleus and cytoplasm, and calculate the ratio as a measure of translocation. This is a clumpy cell type, so studying the settings in primary object identification may be helpful for users interested in the more advanced options that module offers. More about these images can be found at the BBBC.
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This pipeline shows how to identify smaller objects (foci) within larger objects (nuclei) and how to use the Relate module to establish a relationship between the two as well as perform per-object aggregate measurements (such as number of foci per nucleus).
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This example shows an example of object tracking. This pipeline analyzes a time-lapse experiment to identify the cells and track them from frame to frame, which is challenging since the cells are also moving. In addition, this pipeline also extracts metadata from the filename and uses groups the images by metadata in order to independently process several sequences of images and output the measurements of each.
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Pipelines which have appeared in published papers are provided here. Please keep in mind that the listed pipelines are specific to the published assay and that no images are provided.
Example database, configuration file, and images: Single 96-well plate of human cellular images, for use with CellProfiler Analyst 2.0. More sample images and information on the image data can be found on the Broad Bioimage Benchmark Collection (BBBC). Moffat J et al., A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Cell. 2006 Mar 24;124(6):1283-98
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Previous examples (prior to 3.0) can be found on the previous examples page, but we discourage their use because they are now outdated.