Protocols
CSMCFACS Facility User Guide
Sample Preparation
For Analysis
Tubes: Falcon polystyrene tubes 12X75mm (Cat#2008), non-sterile or (Cat#2058), sterile.
Cell Concentration: Approx. 1 x 10^6 cells per mL resuspended in PBS
containing 1% BSA. No phenol red should be used. Minimum sample volume is 500 uL.
For Sorting
Sample Tubes: Falcon polystyrene tubes 12X75mm, (Cat#2058) or Falcon
15mL conical (Cat#2096).
Collection Tubes: Same as above except for 4-way sorting tubes must be
12X75mm. Collection tubes should contain an appropriate volume of media
supplemented with 20% FCS.
Cell Concentration: Approx. 20-25 x 10^6 cells per mL resuspended in PBS
containing 1% BSA. No phenol red should be used. Minimum sample volume is
500 uL. For single cell sorting into 96 well plates concentration should be 1-5 x
10^6 cells per mL.
Cells: Samples should be filtered prior to sorting through filter top tubes Falcon
12X75mm (Cat#2235). For serious cell aggregation problems use DNAse, EDTA
(1 mM) or PBS w/o Ca++ or Mg++ containing 0.1% BSA or FCS.
