The University of Iowa Carver College of Medicine

Flow Cytometry Facility

Flow Cytometry Protocols

Preparing Cell Samples for Flow Cytometry

Detaching Adherent Cells from Tissue Plates without Trypsin

Staining for Viability

Cell Fixation

Disaggregation of Clumped Cells

Immunofluorescence Staining

Absolute Cell Counts

The Facility's flow cytometers do not calculate absolute cell counts (total number of cells per sample). In order to make that calcuation, the total volume of fluid passing through the instrument during data acquisition must be known. Using a known concentration of beads mixed into the cell sample, the fluid volume can be established by the total bead count and then the cells per unit volume can be calculated from the known volume. Several vendors sell beads for this purpose.

Calcium Flux

Apoptosis

DNA Content and Cell Cycle

Intracellular Staining

Links to Other Flow Cytometry Facility Protocols