PMID: 8601699Apr 1, 1996Paper

Identification of F-reticulocytes by two-stage fluorescence image cytometry

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
M L OsterhoutK Horiuchi

Abstract

Precise determination of reticulocytes (young red blood cells) containing fetal hemoglobin (Hb F), F-reticulocytes, is important for assessment of the efficacy of drugs such as hydroxyurea and butyrate in elevating the levels of Hb F in patients with sickle cell disease (SCD) and beta-thalassemia. We developed a reliable and easily applicable method for determining F-reticulocytes using fluorescence image cytometry. Reticulocytes were first identified by preparing a monolayer smear of blood stained by acridine orange. Images of reticulocytes and of all cells were obtained for a selected area on the smear. After removing the acridine orange, cells containing Hb F (F-cells) in the same area were then identified by immunofluorescence. Using images of F-cells, reticulocytes, and all cells for the same fields it was possible to identify F-reticulocytes. To assess the validity of our two-stage staining method, we compared our results with those obtained by traditional methods. There was significant correlation of our method with the conventional immunofluorescence staining method for F-cells (r2 = 0.99; slope = 0.99) and with the accepted brilliant cresyl blue method for reticulocytes (r2 = 0.97; slope = 0.96). Heretofore, the abilit...Continue Reading

Citations

Sep 28, 2001·Journal of Clinical Laboratory Analysis·R S RileyA Tidwell
Jul 4, 2002·Hematology/oncology Clinics of North America·Roger S RileyGina Romagnoli

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