Question: How should I collect blood or bone marrow to preserve neutrophils for single-cell sequencing?
Answer: Vacutainer tubes: Whole blood or bone marrow aspirates from which neutrophils or whole blood leukocytes will be isolated can be collected using vacutainer tubes. Sodium Heparin, Sodium Citrate, K2EDTA, and ACD-A anticoagulants were evaluated and generated comparable single-cell data. 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.
After collection, samples should be transported at Room Temperature (neutrophils do not like cold temperatures) and processed as soon as possible (<24hrs). In-house, our samples were processed <6hrs after collection.
Isolate whole blood leukocytes (including neutrophils) by performing RBC lysis steps (option 1 of our demonstrated protocol Isolation of Leukocytes, Bone Marrow and Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing )
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Fixed RNA Profiling Gene Expression