We have seen that CMFDAd IC21 cells seem to be dying madly. This is a sad sad sad thing, as we all use CMFDA to label cell volume for morphology and cell tracking measurements.
Our control sample of IC21s scanned with transmitted light seems to live fine, especially on the DeltaVision scope in CCR. This indicates to us that the problem is from the combination of intense mercury lamp light and/or CMFDA.
We have found that using Trolox, a water-soluble Vitamin E derivative, allows the cells to live and move under CMFDA/Hg illumination - this has been shown by both Monica Chu on Kinetiscan and Sarah Hopp on DeltaVision.
We have also recently seen that free radical based death is caused by photoinitiators in PEGDA. Different chemicals have different radicals and cause different damage.
We intend to:
Control for everything: Transmitted light scans of CMFDA stained, CMFDA/Trolox Control for illumination: Transmitted light scan of unstained, but with equivalent exposure of Hg lamp light at same wavelengths.
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