Estimation of fetal hemoglobin levels in individual red cells via fluorescence image cytometry
K Horiuchi, ML Osterhout, H Kamma… - … : The Journal of the …, 1995 - Wiley Online Library
K Horiuchi, ML Osterhout, H Kamma, NA Bekoe, KJ Hirokawa
Cytometry: The Journal of the International Society for Analytical …, 1995•Wiley Online LibraryA method for estimating fetal hemoglobin (Hb F) levels in individual red blood cells was
developed. Cell smears were prepared using a slide maker to ensure uniform thickness and
were then stained with immunofluorescence. An antifading gel was applied to preserve a
stable fluorescence. The total fluorescence intensities from the same number of red cells in
different slide specimens correlated with their hemolysate Hb F levels, which were
determined via column chromatography (R= 0.95). Hb F level in individual cells was …
developed. Cell smears were prepared using a slide maker to ensure uniform thickness and
were then stained with immunofluorescence. An antifading gel was applied to preserve a
stable fluorescence. The total fluorescence intensities from the same number of red cells in
different slide specimens correlated with their hemolysate Hb F levels, which were
determined via column chromatography (R= 0.95). Hb F level in individual cells was …
Abstract
A method for estimating fetal hemoglobin (Hb F) levels in individual red blood cells was developed. Cell smears were prepared using a slide maker to ensure uniform thickness and were then stained with immunofluorescence. An antifading gel was applied to preserve a stable fluorescence. The total fluorescence intensities from the same number of red cells in different slide specimens correlated with their hemolysate Hb F levels, which were determined via column chromatography (R = 0.95). Hb F level in individual cells was estimated from fluorescence intensity and cell area, which were determined via image analysis techniques and the hemolysate Hb F level. Blood from a normal subject, a subject with hereditary persistence of fetal hemoglobin, and from sickle cell patients with varying Hb F levels was analyzed. Our analyses showed a wide distribution of Hb F among cells for the normal subject and a gaussian distribution with a peak at the hemolysate Hb F level for the subject with hereditary persistence of fetal hemoglobin. The Hb F distributions were unique to the patients with sickle cell disease. Because Hb F level in individual sickle cells is crucial to the inhibition of cell sickling, the unique Hb F distribution may be important in determining the clinical course of this disease. © 1995 Wiley‐Liss, Inc.
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