Impact of preanalytical variables on granulocytic surface antigen expression: a review.
BACKGROUND
There is a gradual but steady increase in the use of granulocytic surface marker studies to diagnose several inherited and acquired blood and bone marrow disorders. Diagnosis and follow-up of patients with inflammation and infection are other areas of quantitative flow cytometric application. Despite the increased use of flow cytometry to study granulocytes, there seems to be no well-established standards regarding specimen handling for these studies.


METHODS
This review summarizes the effect of preanalytical variables on granulocytic surface markers.


CONCLUSIONS
Storing specimens in sodium heparin at room temperature for up to 72 h seems satisfactory. Other anticoagulants, although acceptable, may have a shorter storage time. Storage time could be prolonged further with the use of some preservation media. Lysed whole blood is the preferred technique. Techniques should avoid major temperature change and excessive manipulation and should maintain storage and methodologic temperature with minimal fluctuation. Fixation before staining with the antibody may result in decreased expression of some surface antigens.
Elghetany
M Tarek
MT
Department of Pathology, University of Texas Medical Branch, Galveston, Texas 77555-0743, USA. melgheta@utmb.edu
Davis
Bruce H
BH
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D000954:Antigens, Surface / Q000096:biosynthesis*; D001793:Blood Preservation / Q000379:methods; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods; D002624:Chemistry, Clinical / Q000379:methods; D005404:Fixatives / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards*; D006098:Granulocytes / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007249:Inflammation; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors
D000925:Anticoagulants; D000954:Antigens, Surface; D002412:Cations; D005404:Fixatives