Abstract

Ethylene diamine tetra acetate (EDTA) added blood is commonly used to perform various analysis like the complete blood count in a hematology laboratory. Prolonged storage in EDTA can cause morphological changes in the cellular components of the blood which has some potential to tamper with the diagnosis and interpretation of results especially during peripheral blood smear reporting and differential counting. This study aims to assess the morphology related variations in WBC and RBC in peripheral smears prepared from EDTA added blood stored at room temperature at different time periods. Peripheral smears were prepared from 20 healthy volunteers after 0 hrs, 2 hrs, 5 hours, 10 hours and 24 hours of storage in EDTA added blood collection containers. Parameters affecting cellular morphology such as crenation, acanthocytes, bite cells, fragmentation and central pallor (for RBCs) and nuclear degeneration, number of lobes in nucleus and vacuolation in cytoplasm (for WBCs) were quantified microscopically in 10 fields/smear after Leishman staining. Students t-test was performed to determine if there was significant morphological variation at different time points and p value less than 0.05 was considered significant. There was significant increase in the number of RBCs with crenation, acanthocytes, bite cells in smears prepared after 2 hours while WBCs showed significant nuclear degeneration and cytoplasmic changes in smears prepared after 1 hour of storage. Maximum cellular distortion was seen in smears prepared after 24 hours of storage at room temperature. Morphological interpretations of RBCS should preferentially be done within 2 hours while for differential count should preferentially be done within 1 hour to minimize errors and misdiagnosis, when blood samples are stored at room temperature.

Keywords: Complete Blood Cell (CBC); Ethylene Diamine Tetra Acetate (EDTA); Crenation; Bite Cells; Acanthocytes; Nuclear Degeneration; Cytoplasmic Vacuolation

References

1. Narasimha Aparna., et al. “Anticoagulant Induced Artefacts in Peripheral Blood Smears”. Indian Journal of Hematology and Blood Transfusion2 (2008): 43-48.
2. Koolwal Dr M S. “Anticoagulant EDTA Induced Storage Effect (Artifacts) on Peripheral Blood Cells”. Journal of Medical Science and Clinical Research1 (2018).
3. Basu Debdatta and Harsha Veluru. “A Study of Storage Related Changes and Effect of Refrigeration on Hematological Parameters and Blood Cell Morphology in EDTA Anticoagulated Blood”. Annals of Pathology and Laboratory Medicine 5 (2019): A289-296.
4. Houwen Berend. “Blood Film Preparation and Staining Procedures”. Clinics in Laboratory Medicine1 (2002): 1-14.
5. Adewoyin A S and B Nwogoh. “Peripheral Blood Film - a Review”. Annals of Ibadan Postgraduate Medicine2 (2014): 71-79.
6. Schapkaitz Elise and Dashini Pillay. “Prolonged Storage-Induced Changes in Haematology Parameters Referred for Testing”. African Journal of Laboratory Medicine1 (2015).
7. Wahed Amer., et al. “Sources of Errors in Hematology and Coagulation”. Hematology and Coagulation (2020).
8. Sukorini Usi., et al. “The Effect of Excessive Disodium Ethylene Diamine Tetraacetic Acid (Na2EDTA) Anticoagulant Concentration on Leukocytes Profile in Peripheral Blood Examination”. Medical Journal of Indonesia3 (2007).
9. Antwi-Baffour Samuel. “Prolong Storage of Blood in EDTA Has an Effect on the Morphology and Osmotic Fragility of Erythrocytes”. International Journal of Biomedical Science and Engineering2 (2013): 20-23.
10. Kafka M and T. Yermiahu. “The Effect of EDTA as an Anticoagulant on the Osmotic Fragility of Erythrocytes”. Clinical and Laboratory Haematology4 (1998).
11. Bartoli V., et al. “Influence of Blood Storage Conditions and Anticoagulants on Results of Blood Cell Filtration Test”. Clinical Hemorheology2 (1986).
12. Zini G. “Stability of Complete Blood Count Parameters with Storage: Toward Defined Specifications for Different Diagnostic Applications”. International Journal of Laboratory Hematology2 (2014).
13. Turhan Turan., et al. “Effects of Storage Conditions on Complete Blood Cell Count Parameters”. Turkish Journal of Biochemistry2 (2011): 165-174.
14. Vives-Corrons Joan Lluis., et al. “Effect of EDTA-Anticoagulated Whole Blood Storage on Cell Morphology Examination: A Need for Standardization”. International Journal of Laboratory Hematology2 (2014): 222-226.
15. Wu Dong wen., et al. “How Long Can We Store Blood Samples: A Systematic Review and Meta-Analysis”. EBioMedicine 24 (2017): 277-285.
16. Sareen Rateesh., et al. “Incubation and Its Effect on Leishman Stain”. Journal of Laboratory Physicians03 (2018): 357-361.

Citation

Citation: Deepa Revi., et al. “Storage Dependent Cellular Changes in Blood Smears Prepared from EDTA Added Venous Blood Samples”.Acta Scientific Medical Sciences 5.6 (2021): 42-46.

Copyright

Copyright: © 2021 Deepa Revi., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.