Related Experiment Video
Updated: May 17, 2025

Characterization of Sickling During Controlled Automated Deoxygenation with Oxygen Gradient Ektacytometry
Published on: November 5, 2019
Time-Dependent EDTA Effect on Leukocyte Differential Count and Morphology Compared to Blood Smear Made by Direct
Anna Irding1, Filip Landgren1, Elisabeth Aardal2
1Department of Clinical Chemistry, Linköping University, Linköping, Sweden.
Introduction:
The impact of ethylenediaminetetraacetic acid (EDTA) on leukocyte count and morphology has led to the current recommendation that smears made from EDTA blood should be prepared within 4 h of sampling. However, previous studies have only been performed on smears with normal cell counts, and smears made from K2EDTA blood have not been studied in comparison with smears made from direct finger prick. The aim of this study was to investigate the time limit for performing a morphological assessment on smears from EDTA blood without compromising the results.
Methods:
Blood samples from 37 patients with known or suspected abnormal leukocyte morphology were selected, with smears from direct finger prick used as controls. Smears were prepared from the original EDTA tube every 4 h up to 24 h after sampling. The smears were evaluated for differential count, morphological assessment, and smear quality scoring.
Results:
No significant difference in cell count was observed on smears made up to 12 h after sampling for neutrophils (p = 0.430), lymphocytes (p = 0.080), monocytes (p = 0.948) eosinophils (p = 0.398), basophils (p = 0.460), and blast cells (p = 0.239) compared to smears made by finger prick. However, smears prepared later than 8 h postsampling showed a significant increase in morphological changes.
Conclusion:
A manual differential leukocyte count can be reliably performed on smears from EDTA blood within 12 h, whereas a comprehensive morphological assessment requires smears to be prepared within 8 h of sampling.
More Related Videos
12:29Live Imaging of Antifungal Activity by Human Primary Neutrophils and Monocytes in Response to A. fumigatus
Published on: April 19, 2017
14:45Enumeration of Major Peripheral Blood Leukocyte Populations for Multicenter Clinical Trials Using a Whole Blood Phenotyping Assay
Published on: September 16, 2012
Related Concept Videos
Effects of EDTA on End-Point Detection Methods
In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a...
EDTA: Direct, Back-, and Displacement Titration
Direct titration involves buffering the metal ion solution to the desired pH and directly titrating with standard EDTA until the endpoint. The optimum pH ensures a large conditional formation constant of metal−EDTA and visibility of the free indicator color in the solution. In addition, auxiliary complexing reagents are used to prevent the precipitation of metal hydroxides...
EDTA: Indirect and Alkalimetric Titration
Masking and Demasking Agents
There are many masking agents, such as cyanide, fluoride, triethanolamine, thiourea, and 2,3-bis(sulfanyl)propan-1-ol (formerly 2,3-dimercapto-1-propanol), with the masking agent chosen based on...