Assessment of a new immunocytochemical technique in HbF‐cell counting by a novel objective evaluation method

Abstract

Summary.A recently developed immunocytochemical technique in HbF-cell counting was assessed by an objective evaluation method. The basic principle of this method is the preparation of aliquots with predetermined HbF-cell (target) values. These aliquots serve as control samples to standardize the HbF-cell measurements by the new immunocytochemical technique, which uses the StreptABComplex/ AP staining procedure (SAP) and visualization under white light. Immunofluorescence optical counts (IF) were performed in parallel with the new technique. A trend of inaccuracy was observed in low target values for both methods. As the level of target values increased, deviations became insignificant (relative accuracy < 8%) with SAP having slightly better results. Linear regression data of the estimated %HbF-cell rates by the two methods versus the target values were very satisfactory for both methods with SAP being slightly better. SAP seems to provide an accurate and reliable alternative for HbF-cell estimation comparable with the classical IF optical count.

Red cell macrocytosis in hypoxemic patients with chronic obstructive pulmonary disease

“…The percentage of F-cells was microscopically determined on peripheral blood smears treated with a 0.4% formaldehyde in acetone solution to allow both erythrocyte fixation and permeabilization. 10 HbF staining was carried out by the classical immunocytochemical streptavidin-biotinylated alkaline phosphatase technique (streptABComplex/ AP, DAKOPATTS a/s Denmark) employing a monoclonal anti-g chain antibody (clone NaM-16-2F4 Bioatlantic, France) as the primary reagent. With these reagents, F-cells stained red; the number and proportion over the whole RBC population was determined by optical microscopy under oil.…”

Immunocytometric quantitation of foeto-maternal haemorrhage with the Abbott Cell-Dyn CD4000 haematology analyser

“…In addition, various alternative approaches such as gel agglutination (Salama et al. , 1998), immunocytochemistry (Tsantes et al. , 2003) and flow cytometry (Patton et al.…”

“…Many different formulae for the determination of FMH by KBT and modifications of the KBT have been described (Corsetti et al, 1987(Corsetti et al, , 1988Bromilow & Duguid, 1997). In addition, various alternative approaches such as gel agglutination (Salama et al, 1998), immunocytochemistry (Tsantes et al, 2003) and flow cytometry (Patton et al, 1990;Thorpe et al, 1994;Johnson et al, 1995;Lloyd-Evans et al, 1996;Davis et al, 1998;Nelson et al, 1998) have been evaluated. Flow cytometric methods using anti-RhD or anti-HbF (gamma-chain) antibodies represent a major improvement on KBT procedures but are often of limited use in routine haematology laboratories because of the need for a dedicated flow cytometer and associated technical expertise.…”