We would like to stain both calcium and endoplasmic reticulum in cancer cells growing in the same plate (dish or flask) and by using only one protocol so that we can merge images to locate calcium (inside ER or in cytoplasm). Is this possible? Can you recommend products-protocols for this imaging process?
I think that the endoplasmic reticulum and calcium staining in cancer cells, growing in the same plate (dish or flask) is not possible. We have to stain separately for both of this. But we it is possible to stain the both with the single protocol. By using ER-Tracker Blue-White DPX (E-12353) and Fluo-3 calcium indicator from Molecular Probes we can stain both the calcium and endoplasmic reticulum with common protocol.
Below is the reference for the common protocol.
3, 3’-Diindolylmethane (DIM) and its derivatives induce apoptosis in pancreatic cancer cells through endoplasmic reticulum stress-dependent upregulation of DR5, Carcinogenesis vol.27 no.4 pp.717–728, 2006
This mesurement is possible - just combine the ER tracker red (EM 610nm) with Fura-2AM (EM 510). If you have adequate filters you can nicely colocalize and additionally you are only staining the intracellular Ca2+. The staining protocols can be combined in one sample.
Dear Vakamullu,
are you interested in staing Ca2+ into the ER or into the cytoplasm? They are two extremely different pools.
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