To help prevent clumping, minimize the time between single cell suspension preparation and loading onto the Tapestri instrument. Additionally, the final cell suspension can be made up to 1 mM EDTA or 5% BSA.
When working with Bone Marrow Mononuclear Cells (BMMCs), the following solution can be used during thawing and washing to help prevent clumping.
BMMC Wash Solution
Trisodium Citrate Dihydrate 22.0 g (75 mM Sodium Citrate)
Citric Acid Monohydrate 8.0 g (40 mM Citric Acid)
Glucose Monohydrate 24.5 g (125 mM Glucose)
Water to 1 liter
No adjustments to pH needed.
Thaw BMMCs with a 1:1 mix of FBS:BMMC Wash Solution, and use the BMMC Wash Solution in lieu of PBS for subsequent wash steps.