Giemsa-stained wet mount based method for reticulocyte quantification: a viable alternative in resource limited or malaria endemic settings

The quantity of circulating reticulocytes is an important indicator of erythropoietic activity in response to a wide range of haematological pathologies. While most modern laboratories use flow cytometry to quantify reticulocytes, most field laboratories still rely on 'subvital' staining....

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Bibliographic Details
Published inPloS one Vol. 8; no. 4; p. e60303
Main Authors Lee, Wenn-Chyau, Russell, Bruce, Lau, Yee-Ling, Fong, Mun-Yik, Chu, Cindy, Sriprawat, Kanlaya, Suwanarusk, Rossarin, Nosten, Francois, Renia, Laurent
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 02.04.2013
Public Library of Science (PLoS)
Subjects
Analysis
Automation
Azure Stains
Biology
Blood
Cytometry
Diagnosis
Erythrocytes
Flow cytometry
Hematology
Humans
Immunology
Infections
Laboratories
Malaria
Malaria - blood
Malaria - diagnosis
Measurement
Medical examination
Medicine
Methods
Methylene blue
Methylene Blue - analogs & derivatives
Microbiology
Microscopy
Parasitology
Plasmodium vivax
Reticulocyte Count - methods
Reticulocytes
Reticulocytes - parasitology
Stains
Stains & staining
Vector-borne diseases
United States
Kuala Lumpur Malaysia
Malaysia
Singapore
Thailand
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Summary:The quantity of circulating reticulocytes is an important indicator of erythropoietic activity in response to a wide range of haematological pathologies. While most modern laboratories use flow cytometry to quantify reticulocytes, most field laboratories still rely on 'subvital' staining. The specialist 'subvital' stains, New Methylene Blue (NMB) and Brilliant Crésyl Blue are often difficult to procure, toxic, and show inconsistencies between batches. Here we demonstrate the utility of Giemsa's stain (commonly used microbiology and parasitology) in a 'subvital' manner to provide an accurate method to visualize and count reticulocytes in blood samples from normal and malaria-infected individuals.
Bibliography:Competing Interests: Bruce Russell, Laurent Renia, and Francois Nosten are all Academic editors of PLOS ONE. The authors agree to abide by the rules, ethical standards, and practices of PLOS; furthermore this does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.
Involved in ethics submission and clinical sampling collection: FN CC. Conceived and designed the experiments: WCL BR YLL LR. Performed the experiments: WCL BR CC KS RS. Analyzed the data: WCL BR YLL CC KS RS FN LR. Contributed reagents/materials/analysis tools: YLL MYF LR FN KS. Wrote the paper: WCL BR YLL MYF CC RS FN LR.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0060303