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Methodology-dependent variations in reticulocyte counts using a manual and two different flow cytometric procedures
A J van Houte1, P C Bartels, M Schoorl
1Department of Clinical Chemistry, Haematology and Immunology, Medical Centre Alkmaar, The Netherlands.
European Journal of Clinical Chemistry and Clinical Biochemistry : Journal of the Forum of European Clinical Chemistry Societies
|November 1, 1994
Summary
Flow cytometry methods using thiazol orange (FACScan) and auramine O (Sysmex) offer higher reproducibility for reticulocyte enumeration compared to the traditional microscopic brilliant cresyl blue method. These advanced techniques provide a more reliable alternative for counting reticulocytes.
Area of Science:
- Hematology
- Clinical Pathology
- Laboratory Medicine
Background:
- Reticulocyte enumeration is crucial for assessing bone marrow erythropoiesis.
- Traditional microscopic methods using brilliant cresyl blue are labor-intensive and have lower reproducibility.
- Flow cytometry offers potential for more accurate and efficient reticulocyte counting.
Purpose of the Study:
- To compare the accuracy and reproducibility of microscopic reticulocyte counting with two flow cytometric methods: thiazol orange (FACScan) and auramine O (Sysmex).
- To establish reference ranges for reticulocyte counts using each method.
- To evaluate the correlation between microscopic and flow cytometric techniques.
Main Methods:
- Comparison of microscopic brilliant cresyl blue method with FACScan (thiazol orange) and Sysmex (auramine O) flow cytometry for reticulocyte enumeration.
- Intra-assay coefficients of variation were calculated for each method.
- Correlation coefficients (r) were determined between the methods.
- Reference ranges (2.5th and 97.5th percentile) were established for each method.
Main Results:
- Flow cytometry methods demonstrated significantly higher reproducibility (mean intra-assay CV of 5%) compared to the microscopic method (33.9%).
- Poor correlation was observed between microscopic and flow cytometric methods (r = 0.61 for FACScan, r = 0.57 for Sysmex).
- A satisfactory correlation was found between the two flow cytometric methods (r = 0.79).
- The Sysmex R-3000 method yielded the narrowest reference range (8-18/1000).
Conclusions:
- Flow cytometric analysis of reticulocytes offers superior reproducibility over the microscopic enumeration method.
- The Sysmex R-3000 method provides a reliable and reproducible approach for reticulocyte counting with a narrow reference range.
- Flow cytometry is an attractive alternative for reticulocyte enumeration due to its enhanced accuracy and efficiency.