Cellprofiler6/26/2023 In 5 images of well separated Jurkat cells in culture, manual analysis identified 236 cells with profile areas of (mean ± s.d.) 6608 ± 2466 pixels, while CellProfiler identified 185 cells with profile areas of 6190 ± 3175 pixels. The jpg files were converted to tif files for Scion Image. The pipeline was validated against manual analysis using Scion Image software (Scion Corporation, Frederick, MD, USA). Images were saved as jpg files for analysis by CellProfiler. Phase contrast images were taken of Jurkat cells in culture with a Nikon Diaphot inverted microscope fitted with a Nikon D-40 digital camera. The pipeline also incorporates upper and lower size cutoffs to rule out cell clusters and debris. CellProfiler has an edge finder module that we have exploited in developing a pipeline that can automatically measure the sizes of cells in a large number of phase contrast images. On the other hand, phase contrast images tend not to have a high contrast between the cell interior and the background, but show an edge at the boundary of the cell. CellProfiler (1) is a software package that was designed for the automated analysis of fluorescence images, which feature high contrast between labelled cells and the background, enabling the software to identify cells in the image. We aimed to develop a cheap and easy method for measuring cell size that could generate pilot data to be followed up with more sophisticated methods. The method of choice is usually flow cytometry, which involves expensive equipment and high running costs that may tend to prohibit speculative pilot studies. Studying cell volume control requires an efficient method of measuring cell size in populations of cells. Particularly in the case of apoptosis, this mechanism is not well understood. Change in cell volume is a fundamental and probably essential feature of both cell division and apoptosis, so it is important to understand mechanisms of cell volume control. Upsetting this balance can lead to a variety of diseases.
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