Dihydrorhodamine-123 flow cytometry method: time for substantial revision in technical procedure

Author:

Zavvar Mahdi12ORCID,Zargaran Sina23ORCID,Baghdadi Hamed24ORCID,Poopak Peyvand2,Poopak Amir Hossein25,Nabatchian Fariba1,Fatahi Yousef67,Khosravipour Gelareh2,Poopak Behzad28ORCID

Affiliation:

1. Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Tehran University of Medical Sciences , Tehran , Iran

2. Payvand Clinical and Specialty Laboratory , Tehran , Iran

3. Faculty of Dentistry, University of Toronto , Ontario , Canada

4. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University , Tehran , Iran

5. Cancer Research Center, Tehran University of Medical Sciences , Tehran , Iran

6. Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences , Tehran , Iran

7. Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences , Tehran , Iran

8. Islamic Azad University, Tehran Medical Sciences Branch , Tehran , Iran

Abstract

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.

Funder

Tehran University of Medical Sciences

Publisher

Oxford University Press (OUP)

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