Dear Fellow Researchers,
My colleagues and I ran into some hurdles trying to do intracellular staining for flow cytometry. We are trying to stain human astrocytes with fluorescent GFAP antibody, and analyze the cells on the FACScan. We do the fixation and permeabilization and then staining as suggested by the manufacturer, but are not seeing anything. Does anyone have any suggestions? I have heard that it's much harder to do suspension intracellular staining on fibroblast-body-type cells because they don't stretch out in suspension as well as on slides, which makes cytokeratine type staining difficult, is this true? Is there any special tricks to intracellular staining for cells in suspension? I'd appreciate any comment, thanks so much!