Neutrophils are the most abundant white blood cells in circulation. They are the first responders of the innate immune system that release cytotoxic compounds to kill bacteria and phagocytose foreign particles. Analysis of neutrophils and granulocytes is challenging in single cell transcriptomic data due to the low RNA content in these cells along with relatively high levels of RNases and other inhibitory compounds. Furthermore, neutrophils are sensitive to degradation after collection, requiring careful sample handling for preservation. This Technical Note describes the successful isolation and analysis of neutrophil populations from whole blood.