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  • Protocol for one-step selective lysis of red blood cells and platelets with long-term preservation of white blood cells (human) at ambient temperature.

Protocol for one-step selective lysis of red blood cells and platelets with long-term preservation of white blood cells (human) at ambient temperature.

STAR protocols (2021-09-28)
Rajesh K Chaurasia, Kapil B Shirsath, Balvinder K Sapra
ABSTRACT

Current protocols for storage of white blood cells (WBCs) rely on constant refrigeration. The protocol described below explains the preparation of a fixative combination saline (FCS) formulation, which allows fixation of human WBCs and lysis of red blood cells and platelets (at ambient temperature, 4-35oC) in whole blood samples in one step. FCS can be used for storing and transporting blood at ambient temperatures for up to 4 months, without altering the nuclear morphology and genomic integrity of WBCs.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Acetic acid, glacial, ReagentPlus®, ≥99%
Sigma-Aldrich
Potassium chloride, Molecular Biology, ≥99.0%
Sigma-Aldrich
TWEEN® 20, Molecular Biology, viscous liquid
Sigma-Aldrich
Sodium phosphate dibasic, ACS reagent, ≥99.0%
Sigma-Aldrich
Triton X-100, Molecular Biology