Guava easycyte 6 2l capillary cytometer

After appropriate treatments, cells were loaded with CellTracker Green (200 μmole/L) for 30 min on ice, followed by two washes in cold PBS. Fluorescent emission at 525 nm was analyzed by flow cytometry in a Guava EasyCyte 6-2L capillary cytometer (Merck, Millipore) using a 488-nm argon ion excitation laser. Results are expressed as MFI.

After appropriate treatments, cells were washed twice in PBS and then resuspended in 100 μl Annexin binding buffer containing a 1:100 dilution of Annexin V–FITC and 50 nmol/L tetramethyl-rhodamine methyl ester; where appropriate for MSC coculture experiments, a 1:100 dilution of anti-CD90 APC-conjugated antibody was added. CD90 was used to discriminate MSC (positive) from leukemia cells (negative). In some experiments viable leukemia cells were determined by staining with propidium iodide (Sigma), and viable leukemia progenitors were assessed after staining with anti-CD34 APC-conjugated antibody. Cells were then analyzed by flow cytometry in a Guava EasyCyte 6-2L capillary cytometer (Merck, Millipore) or a FACSCalibur cytometer (BD Biosciences) using a 488-nm argon ion and 633-nmHeNe excitation lasers.