HCT116 cells were monitored with PWS for a period of 24 hours. Images were captured every 15 min. Ten cells that could be observed dividing during the 24-hour period were identified and chosen for analysis. D was tracked for 3 hours before cell division and at least 6 hours after cell division.

To analyze spatial correlations between progeny cells and their progenitors, histograms of D at all pixels within the nucleus were compared. First, each cell was analyzed at each time point. A histogram of D was calculated with 10 evenly spaced bins with widths of 0.15. Each histogram was analyzed by being normalized by the average histogram of all cells at the same time point. For example, a histogram of cell #1, 3 hours before cell division, was normalized by the average of all cells (N = 10) histograms 3 hours before cell division. Therefore, these normalized histogram ratios focused on each cell’s specific deviation from the mean of the population at a specific time point. See fig. S7 for a step-by-step explanation for how histograms were calculated and normalized. The Pearson correlation coefficient was calculated in MATLAB with the “corrcoef” function comparing every pair of progeny cells at each time point. Also, all progeny cells at 3 hours after division were compared to all progenitor cells at 3 hours before division. Three hours was chosen to compare cells before and after division at relatively stable time points (i.e., not during cell detachment or nuclear splitting).

To analyze temporal correlations in averaged nuclear D, the Pearson correlation coefficient was calculated in MATLAB with the corrcoef function to compare time-dependent changes in D between each of the progeny cells (after division). D values measured for a 2-hour segment was compared between all progeny cells and progenitors. The 2-hour time period was 1 hour before (for progenitors) and after (for progeny cells) division. A 1-hour “buffer” around the time of cell division was chosen since the measured D was highly dynamic during this time and subject to artifacts since the cell was in the process of partially detaching from the glass substrate.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.