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Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay


Hussain, Husniza and Santhana, Raj L. and Ahmad, Syahida and Abd Razak, Mohd Fuat and Wan Mohamud, Wan Nazaimoon and Bakar, Jamilah and Mohd. Ghazali, Hasanah and Yildiz, Fatih (2019) Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay. Cogent Food & Agriculture, 5. pp. 1-9. ISSN 2331-1932


In vitro cell viability tests are usually done using 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. A new spectrofluorometry method was developed using acridine orange (AO) and propidium iodide (PI) using multi-label microplate reader. Nine biogenic amines (BAs) [histamine (HIM), putrescine (PUT), cadaverine (CAD), 2-phenylethylamine (PHM), tyramine (TYM), tryptamine (TPM), spermine (SPM), spermidine (SPD) and agmatine (AGM)] were exposed to RAW 264.7 macrophage in singles at 37°C with 5% carbon dioxide supplementation for 18–24 hours and cell viability was determined using MTS method and AO/PI developed method using dual-spectrofluorometry. Based on MTS assay, SPM and SPD were found to be cytotoxic and it was supported by AO/PI assay. The precedence of disintegration in the nucleus rather than mitochondria upon cell non-viability was also supported by transmission electron microscopy (TEM). The results showed that AO/PI method could be used as an alternative method to determine cytotoxicity besides usual usage in confocal microscopy.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Food Science and Technology
DOI Number: https://doi.org/10.1080/23311932.2019.1582398
Publisher: Taylor & Francis
Keywords: Acridine orange; Propidium iodide; Dual-spectrofluorometry; Cell viability
Depositing User: Ms. Nuraida Ibrahim
Date Deposited: 10 Sep 2020 03:11
Last Modified: 10 Sep 2020 03:11
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1080/23311932.2019.1582398
URI: http://psasir.upm.edu.my/id/eprint/81120
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