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Cell Cycle studies are commonly performed through flow cytometry in order to obtain the proportion of cells at each phase of the cell cycle. But what about fine cellular analysis or concomitant protein localization analysis. Go one step beyond conventional methods with Pathfinder™ Cellscan Image Cytometer. Multispectral microscope-based analysis enables quantification of huge populations of cells, whilst maintaining information, as well as image, for each individual cell. The following example demonstrates just some of the Pathfinder™ Image.
Cytometer capabilities:
Nuclei of HeLa cells in culture were counterstained with DAPI, mitosis were labeled in red using an antibody, cells in anaphase and telophase were labeled in green and nuclei were automatically detected:
For each individual cell, the amount of DNA in the nucleus (total blue signal intensity) was measured, as well as the red and green intensities. Again, all the calculated values can be plotted in histograms (with optional multi-gaussian fit), multi-histograms or scatter-plots, and sub-populations can be isolated on those plots. All cells belonging to an isolated sub-population are then highlighted both on the images and in the results table with automated re-calculation of the statistics on all the parameters for the sub-population of interest.
Moreover, a simple mouse click enables automated re-postioning of the cells of interest under the eyepiece of the microscope. The scatter plot below shows the distribution of the ratio anaphase/telophase (green) vs mitosis marker (red) signal intensities among DNA content of the cells (blue signal) for this experiment, demonstrating the ability to isolate sub-populations and have direct one-click access to the cells belonging to this supopulation (labeled with a pink dot).
Performances:
Time for capture and analysis of the samples depends on the quality of the samples and the complexity of the analysis, but in most cases, the full process takes less than 10 minutes for a 96-wells plate and may be as fast as 5 minutes per plate.
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