User:Easilva/Notebook/Wayne/Entry Base

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Checking efficiency of Live/Dead staining Main project page

20130217

  • 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...



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