Live-Cell Analysis of Cell Subsets and Heterogeneity: Your Questions Answered
An expert cellular biologist answers your questions on cell subset identification in studying heterogeneous populations of cells
8 Oct 2018
Expert insights
Cell-by-cell analysis can help characterize cell subsets and monitor cell behavior in real time. In a recent webinar, Dr. Clare Szybut presented how cell-by-cell analysis enables monitoring and quantification of dynamic phenotypic changes in heterogeneous cell populations and reveals the exciting new software that can make it possible.
If you didn’t catch it live, no problem, we’ve got you covered. You can watch the webinar on-demand here >>
Watch Webinar NowThank you for all your questions. Here are some of the highlights from the Q&A session, which took place over Twitter:
Q: Can the data for individual cells be exported for analysis/visualization in other packages?
A: Yes. Data can be readily exported to packages such as Excel. Due to the large file sizes, each time point is exported as a separate file.
Q: Is the Fab-based antibody labeling approach suitable for use on other platforms, such as flow cytometry.
A: Yes. The phenotyping validation data of PBMCs was achieved using the FabFluor488-Ab labeled cells on a flow cytometry instrument.
Q: Will the software module enable segmentation of adherent cell types?
A: No. At present, the module has not been developed or validated for use with adherent cells. However, that is an obvious area of interest.
Q: What is the mask?
A: The mask refers to the segmentation of the phase image enabling each cell to be identified.
Q: Have you used this technology for any primary cells?
A: Yes, we have used live-cell imaging extensively with PBMC and isolated cell types from the blood.
Q: If I have two cell lines, one labeled with GFP and the other RFP, can this system detect the two cell types, monitor growth and roundness?
A: Yes, this would be an ideal set-up for this new software module.
Q: What is special about these antibodies, apart from being non-toxic?
A: There is nothing special about them, but they have been labeled with IncuCyte FabFluor-488 for detection. We do recommend you purchase low azide containing antibodies.
Q: What do you do to keep suspension cells centered in the wells? We have problems with cells moving towards the edges of the wells.
A: We have observed similar issues. Routinely, we coat our plates with PLO to reduce this issue. Please see the Essen BioScience website for full protocols and details of the use of non-adherent cells with IncuCyte.
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