Red Blood Cell Biology Laboratory

Methods

Blood fractionation
Blood leukoreduction
Colorimetric Elisa
Confocal Laser Scanning Microscope (CLSM)
Conventional electron microscopy (TEM, SEM)
Density fractionation of RBCs (in vivo aging)
Flow cytometry
Fluorometry
Immunoblotting
Immunofluorescence microscopy
Immunogold electron microscopy (unicryl acrylic resins)
Immunoprecipitation
In vitro model of transfusion (RBC reconstitution with plasma at body ToC)
Isolation of RBCs and EVs
Subcellular fractionation of RBCs (preparation of ghosts, cytosol, membrane fractions/skeleton)
Statistical and network analyses

Assays for measurement of:

Caspase activation (bulk, in situ)
EV-associated hemolysis
Glutathione (GSH)
Hemolysis and cellular fragilities (upon storage, osmotic, mechanical and oxidative stresses)
Oxidative and Ca2+ stress (intracellular ROS/Ca2+ accumulation, protein oxidative modifications, lipid peroxidation)
Protein identification and quantitation (red cell fractions and EV phenotyping)
Proteasome activity
PS exposure
RBC shape distribution (spherocytosis, stomatocytosis, echinocytosis)
Total Antioxidant Capacity (TAC) of plasma (FRAP assay) (UA-dependent/independent)