No cells were detected to be attached to T25 flask since last Friday;
T25 was trashed today;
T75 (60% confluent HMVEC-D) flask was labeled with Live/Dead - Protocol: Cells were washed twice with PBS; 5 ul of both working solution of Calcein and Ethidium (2 uM and 4uM) were added to 5 mL of PBS; Cells were incubated for 45 min with 3 ml [from the solution] (plus fresh 7 mL of PBS) of labeling solution; washed twice with PBS and fresh media was used;
Cells were visualized under the microscope and pictures taken and stored in the microscope computer under wayne folder;
Still not clear and relatively weak signal from ethidium, could happen that cells are just very viable...