I used two conditions. In one well I stained the cells using only Calcein AM & Hoechst and the other well was stained with Ethidium Homodimer-1 and hoechst.
The cells stained with only Calcein-AM & Hoechst is showing signal in red and the cells which were stained only with Ethidium Homodimer-1 and hoechst showed signal in green color. I am confused now. Why will they signal in those color even when the stains are not added?
Calcein AM is a non-fluorescent, uncharged molecule that can pass through the plasma membrane of viable cells. Once inside the cells, Calcein AM is hydrolyzed by intracellular esterases into the green-fluorescent. Ethidium Homodimer-1 in impermeant dye and Hoechst dyes are cell membrane-permeant.
When we detected live, apoptotic dying and dead cells, we used Calcein AM to show live cells, Apoptotic-dead cell tag (surface staining) to show apoptotic dying and dead cells, and fixable viability dye or DAPI to dead cells. Please visit www.biomagis.com
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