Importantly, fluorescence is, in principle, quantitative in that intensity of fluorescence from each position in a sample is proportional to the abundance of the fluorescent moiety in that region of the sample.
We believe this method will help other non-image processing specialists use microscopy for quantitative image analysis.įluorescence microscopy is the method of choice to visualize specific cellular organelles, proteins, or nucleic acids with high sensitivity and selectivity. Our method is straightforward and easily applicable using our staining protocol. In a second application, we quantified changes in the projected cell area of CHO cells upon lowering the incubation temperature, a common stimulus to increase protein production in biotechnology applications, and found a stark decrease in cell area. Using this approach, we quantified changes in the projected cell area, circularity, and aspect ratio of THP-1 cells differentiating from monocytes to macrophages, observing significant cell growth and a transition from circular to elongated form. 86% of the evaluated cells were segmented correctly, and the average intersection over union score of detected segmentation frames to manually segmented cells was above 0.83. Resultsīased on an evaluation with HeLa cells compared to human counting, our algorithm reached accuracy levels above 92% and sensitivity levels of 94%. In our method, the “leaky” fluorescence from the DNA stain DRAQ5 is used for automated nucleus detection and 2D cell segmentation.
#Imagej automatic cell counter software
In this work, we present a fast, customizable, and unsupervised cell segmentation method that is based solely on Fiji (is just ImageJ)®, one of the most commonly used open-source software packages for microscopy analysis. Image segmentation and quantification are essential steps in quantitative cellular analysis.
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