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Dihydrorhodamine-123 Flow Cytometry Method: Time for Substantial Revision in Technical Procedure Publisher Pubmed



Zavvar M1, 2 ; Zargaran S2, 3 ; Baghdadi H2, 4 ; Poopak P2 ; Poopak AH2, 5 ; Nabatchian F1 ; Fatahi Y6, 7 ; Khosravipour G2 ; Poopak B2, 8
Authors
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Authors Affiliations
  1. 1. Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Payvand Clinical and Specialty Laboratory, Tehran, Iran
  3. 3. Faculty of Dentistry, University of Toronto, ON, Canada
  4. 4. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
  5. 5. Cancer Research Center, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  7. 7. Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  8. 8. Islamic Azad University, Tehran Medical Sciences Branch, Tehran, Iran

Source: Laboratory Medicine Published:2025


Abstract

The dihydrorhodamine 123 assay is generally applied to measure the production of intracellular reactive oxygen species in neutrophils using flow cytometry and is considered a diagnostic evaluation for chronic granulomatous disease. In fact, there is a broad range of variables that can directly or indirectly affect test results, either individually or collectively. It is therefore crucial to identify the ideal requirements to achieve reliable results as well as using these requirements to provide standard operating procedures that should be taken into account. Therefore, we focus on aligning optimum results by comparing preanalytical and analytical phases that influence test results, such as the effect of various anticoagulants, transport and maintaining temperature (24°C or 4°C) of samples, test prime run time, appropriate solution concentrations, and effect of incubation temperature (24°C or 37°C) during the test run. © The Author(s) 2024. Published by Oxford University Press on behalf of American Society for Clinical Pathology. All rights reserved.