A clinical research consortium sponsored by NIAID and JDRF
The Department of Flow & Image Cytometry at Roswell Park Cancer Institute serves as a core facility for the ITN, providing comprehensive flow cytometric immunophenotyping services.
Blood specimens received by RPCI blood are processed and stained with a selection of common and trial-specific 5 color monoclonal antibody (mAb) cocktails. The cocktails are designed to identify and quantitate most lymphocyte and many myeloid subsets. On average, each patient sample is stained with 10 different, 5 color mAb cocktails. Following staining, data is acquired on FACSCanto (BD Biosciences, San Jose, CA) flow cytometers and analyzed using WinList software (Verity Software House, Topsham, ME).
Other services include assessment of specimen viability, DNA cell cycle, apoptosis (either Annexin + PI or TUNEL), intracellular proteins and cytokines (activation with polyclonal stimulants, peptide or overlapping peptide pools), HLA class I tetramers, proliferation by dye dilution (PKH reagents or CFSE), and soluble bead array for chemokines and cytokines.
