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. elegans and extract measurements on a per-worm basis. These pipelines have been developed for high-throughput screens on C. CellProfiler can be used to analyze the resulting images from imaging flow cytometry, whether brightfield, darkfield, or fluorescence. Imaging flow cytometry combines the high-throughput capabilities of conventional flow cytometry with single-cell imaging. This quantification will enable you to compare the number of dots per cell in wild-type and mutant conditions as well as to measure how the number of dots per cell depends on primary antibody concentration.
CELLPROFILER MEASURE OBJECT SIZE SHAPE HOW TO
In this practical you will learn how to set up an automated CellProfiler image analysis pipeline that will (1) identify individual cells in images, based on a nuclear stain, (2) identify dot-like signals, and (3) count the number of dots per cell and output this information to a spreadsheet. This example shows how the object identifcation and RelateObjects modules are used to measure the degree of overlap between two fluorescent channels. 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 CorrectIlluminationCalculate and CorrectIlluminationApply modules are used to compensate for the non-uniformities in illumination often present in microscopy images. Illumination correction is often important for both accurate segmentation and for intensity measurements. Rather than identifying individual cells, this pipeline quantifies the area occupied by the tissue sample. In this example, cells are grown as a tissue monolayer. 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. Tissue samples often have irregularly shaped cells with adjacent edges. This pipeline identifies patches of yeast growing in a 96 well plate, serving as an introduction to the grid defintion and identification modules. 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. The example identifies uniformly round objects, in this case, yeast colonies growing on a dish. This pipeline demonstrates how to classify and count objects on the basis of their measured features. This pipeline shows how to do both of these tasks, and demonstrates how various modules may be used to accomplish the same result. In addition to cellular object and feature identification, these pipelines include some of the more specialized modules in CellProfiler for image pre-processing or measurement.Ĭell/particle counting and scoring the percentage of stained objectsĬellProfiler is commonly used to count cells or other objects as well as percent-positives, by measuring the per-cell staining intensity. Also shown is a silver-stained comet example in which the percentage of DNA contained in the tail is calculated. Also, illumination correction is used to reduce background fluorescence prior to measurement. Here, the measurement of interest is the length and intensity of the comet tail. This is a simple example of a DNA damage assay using single cell gel electrophoresis.
This pipeline demonstrates how to identify these clumpy cells and obtain morphological, intensity and texture measurements.Ī simple pipeline that identifies and counts tumors in a mouse lung, and then measures their size. In contrast to the HT29 cells, Drosophila Kc167 cells are a highly textured and clumpy cell type. This pipeline demonstrates how to accurately identify these cells and how to measurements cellular parameters such as morphology, count, intensity and texture. Human HT29 cells are fairly smooth and elliptical.
These pipelines are made for simple cellular and tissue image assays, and include some basic measurements. Sequencing RNA molecules in situ combining CellProfiler with ImageJ plugins.Object Tracking and Metadata Management.Human cytoplasm-nucleus translocation assay (SBS Bioimage).Human cytoplasm-nucleus translocation assay (SBS Vitra).Untangle worms and make measurements bright-field staining pattern phenotype.Straighten worms and extract intensity measurements using a low-resolution atlas.Cell/particle counting and scoring the percentage of stained objects.OLD CellProfiler example images and pipelines (Prior to 4.0) These examples are outdated, please see our current Examples page for the updated versions.