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

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 bio...

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Main Authors: Hussain, Husniza, Santhana, Raj L., Ahmad, Syahida, Abd Razak, Mohd Fuat, Wan Mohamud, Wan Nazaimoon, Bakar, Jamilah, Mohd. Ghazali, Hasanah, Yildiz, Fatih
Format: Article
Language:English
Published: Taylor & Francis 2019
Online Access:http://psasir.upm.edu.my/id/eprint/81120/
http://psasir.upm.edu.my/id/eprint/81120/1/CELL.pdf
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author Hussain, Husniza
Santhana, Raj L.
Ahmad, Syahida
Abd Razak, Mohd Fuat
Wan Mohamud, Wan Nazaimoon
Bakar, Jamilah
Mohd. Ghazali, Hasanah
Yildiz, Fatih
author_facet Hussain, Husniza
Santhana, Raj L.
Ahmad, Syahida
Abd Razak, Mohd Fuat
Wan Mohamud, Wan Nazaimoon
Bakar, Jamilah
Mohd. Ghazali, Hasanah
Yildiz, Fatih
author_sort Hussain, Husniza
building UPM Institutional Repository
collection Online Access
description 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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spelling upm-811202020-09-10T03:11:20Z http://psasir.upm.edu.my/id/eprint/81120/ Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay Hussain, Husniza Santhana, Raj L. Ahmad, Syahida Abd Razak, Mohd Fuat Wan Mohamud, Wan Nazaimoon Bakar, Jamilah Mohd. Ghazali, Hasanah Yildiz, Fatih 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. Taylor & Francis 2019 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/81120/1/CELL.pdf 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 https://www.tandfonline.com/loi/oafa20 10.1080/23311932.2019.1582398
spellingShingle Hussain, Husniza
Santhana, Raj L.
Ahmad, Syahida
Abd Razak, Mohd Fuat
Wan Mohamud, Wan Nazaimoon
Bakar, Jamilah
Mohd. Ghazali, Hasanah
Yildiz, Fatih
Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title_full Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title_fullStr Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title_full_unstemmed Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title_short Determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
title_sort determination of cell viability using acridine orange/propidium iodide dual-spectrofluorometry assay
url http://psasir.upm.edu.my/id/eprint/81120/
http://psasir.upm.edu.my/id/eprint/81120/
http://psasir.upm.edu.my/id/eprint/81120/
http://psasir.upm.edu.my/id/eprint/81120/1/CELL.pdf