Quantitative Analysis of Cultured Cells: High-End Solution for Ready-To-Publish Data

Cell cultures are often used for the investigation of different research questions, such as cell growth, proliferation, toxicity studies, and kinetics of biological samples. In addition to methods that provide information about the presence and quantity of metabolism products, cell culture imaging/image analysis provides information on the count, shape, and expression of various markers of fixed cells (Scholz et al, 2019). Depending on the scale of the study and cell line, manual cell culture analysis can often be a time-consuming process, especially in cell dense cultures. The demand for automatization of analytical processes calls for new solutions to enable fast and reproducible data output.

The following case study uses the StrataQuest image analysis solution for the analysis of a fixed and IF-processed monolayer of cultured cells. It shows how StrataQuest enables a simple and automated pipeline for cell identification and marker detection. The aim of this project was to detect and count the nuclei and the cytoplasm, and in addition, to assess the intensity of the red marker.

intro 2 min

(a) shows the original fluorescent cell culture image, where cytoplasm was stained for a red marker and nuclei for DAPI (blue). (b) and (c) represent their gray channels, whereby the nuclei marker channel was later used to identify cell nuclei in the turquoise mask, either on (d) gray or (e) overlay image. To detect cell cytoplasm, a special algorithm was applied: cytoplasm mask starts from the nucleus mask and grows until the signal intensity of the cytoplasmic marker drops. (g) shows the combined masks.

Cell culture pic2

As an output of the analysis, a single FACS-like scattergram (h, left) can provide information on the number of cells, where a cytoplasmatic marker is expressed with the highest intensity (n=14), as well as the overall number of cells in the image (n=373). Cells with the highest intensity of the marker can be also visualized via a backward connection (h, right).

StrataQuest can be used to detect and quantify not only cells, but also cellular substructures. Check our StrataQuest App Center for the variety of applications developed for specific research projects. We also offer custom App development in line with your needs and wishes – contact our team today to find out more.

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