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Potential pitfalls of comparative measurements of reticulocytes with flow cytometry and microscopy in prematures and infants

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Published/Copyright: September 21, 2011
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 42 Issue 10

Abstract

Reticulocyte counting by flow cytometry (Bayer H*3, ADVIA 120) in blood of prematures, infants and children > 1 year of age was compared with microscopic counting under research conditions (9000 counted red blood cells per slide). While in children > 1 year a good concordance of both methods was observed, 2.3–2.4-fold higher values were obtained in neonates by microscopy (Brilliant Cresyl Blue stain, 0.5%). However, another laboratory found good agreement between H*3-counting and microscopy in samples also obtained from neonates using the same methods. Despite very similar results for all age groups in comparative flow cytometry measurements in both laboratories, counting of smears from neonates differed, showing an approximately 2.3-fold larger amount of reticulocytes in our laboratory. The reason for these observations was a greater enlargement (1250-fold) used routinely in our laboratory compared with 800-fold in the other one. Thus very mature reticulocytes frequently found in neonates could only be detected using a 1250-fold enlargement. Similarly, the low concentration of the colouring matter used in the H*3 (0.0005% oxazin or 0.001% ADVIA 120) is obviously not sufficient for detection of mature reticulocytes. Therefore, it is important to consider this phenomenon and to standardise microscopic enlargement, especially for comparisons in multicentre studies.

Keywords: flow cytometry; infants; microscopy; neonates; preterms; reticulocytes

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Received: 2004-6-30
Accepted: 2004-8-20
Published Online: 2011-9-21
Published in Print: 2004-10-1

© Walter de Gruyter

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https://doi.org/10.1515/CCLM.2004.234
Keywords for this article
flow cytometry; infants; microscopy; neonates; preterms; reticulocytes

Articles in the same Issue

  1. The significance of serum γ-glutamyltransferase in cardiovascular diseases
  2. Anti-tissue transglutaminase antibodies in inflammatory bowel disease: new evidence
  3. Expression level of MDR1 message in peripheral blood leukocytes from healthy adults: a competitive nucleic acid sequence-based amplification assay for its determination
  4. Characteristics of prolinase against various iminodipeptides in erythrocyte lysates from a normal human and a patient with prolidase deficiency
  5. Alteration of homocysteine catabolism in pre-eclampsia, HELLP syndrome and placental insufficiency
  6. Immune system-mediated endothelial damage is associated with NO and antioxidant system disorders
  7. Bone marrow microvascular density and angiogenic growth factors in multiple myeloma
  8. The insulin-like growth factor system in the circulation of patients with viral infections
  9. Time course of systemic markers of inflammation in patients presenting with acute coronary syndromes
  10. Age- and sex-related reference values for serum insulin concentration and its biological determinants in a French healthy population. The STANISLAS cohort
  11. Potential pitfalls of comparative measurements of reticulocytes with flow cytometry and microscopy in prematures and infants
  12. Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay
  13. Analytical and diagnostic accuracy of the EliA™ automated enzyme fluoroimmunoassay for antineutrophil cytoplasmic autoantibody detection
  14. A particle-enhanced turbidimetric immunoassay for quantitative determination of orosomucoid in urine: development, validation and reference values
  15. Analytical performance and clinical results of a fully automated MEIA system for brain natriuretic peptide assay: comparison with a point of care testing method
  16. Multicentre performance evaluation of the E170 Module for MODULAR ANALYTICS
  17. Congress of Clinical Chemistry and Laboratory Medicine Annual meeting of the German United Society for Clinical Chemistry and Laboratory Medicine (DGKL), Düsseldorf, Germany, November 22-24, 2004
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