Question: How should I collect blood to isolate PBMCs for single-cell sequencing?
Answer : There are a number of options for blood collection tubes:
Vacutainer tubes: Whole blood can be collected using vacutainer tubes. In-house we have evaluated Sodium Heparin, Sodium Citrate, K2EDTA, and ACD-A anticoagulants and the single-cell data generated is comparable. Users looking to isolate PBMCs can collect samples in these tubes and perform PBMC isolation using a ficoll gradient and SepMate tube ( follow option 3 of our demonstrated protocol Isolation of Leukocytes, Bone Marrow and Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing). Vacutainer tubes are also a good option for users who may want to do multiple assays (i.e. Flow, CytoF, ELISA) from the same tube of blood.
CPT tubes: Whole blood from which PBMCs (no granulocytes) are to be isolated may benefit from the use of CPT tubes. CPT tubes with Sodium Heparin and Sodium Citrate anticoagulants were evaluated in-house and generated comparable single-cell data. These tubes already contain a density gradient. After blood collection, tubes are spun to separate the sample into layers and PBMCs are isolated ( follow option 2 of our demonstrated protocol Isolation of Leukocytes, Bone Marrow and Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing). CPT tubes are a good option for beginners as the density separation is only a 1-step centrifugation process, and only 1 tube is needed for both collection and PBMC isolation.
Products: Single Cell Gene Expression, Single Cell Immune Profiling.