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Routine Hemoglobinopathy



Isoelectric focusing (IEF)
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Routine Hemoglobinopathy

 

Red Blood Cell Pit Count

Counts of  pitted red blood cells (RBC) can be used to assess splenic reticuloendothelial function in patients with sickle cell disease.  There is an inverse relationship between the pit count and the degree of splenic dysfunction. Less than 2% of RBCs contain pits in individuals with normal splenic function, whereas up to 50% of RBCs may demonstrate pitting in individuals with functional asplenia.

Erythrocytes are fixed with 1% glutaraldeyde and examined under phase contrast microscopy to visualize and manually count pitting as an estimate of the number of pitted red cells.   (Ref. Fatunde, O.J. and Scott, R.B. 1986 J Ped Hem Onc )

 
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Pit

Figure 1. Peripheral blood of the propositus, showing irreversibly sickled cells and pitted red cells. A wet prearation is shown under a nomarski objective (x400).

Ref: Witkowska, HE, et al. Sickle Cell Disease in a Patient with Sickle Cell Trait and Compound Heterozygosity for Hemoglobin S and Hemoglobin Quebed-CHORI. NEJM. 1991. Vol. 325 No. 16. Pp 1150-1154.