Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity

This study presents a straightforward and reliable method for determining the viability of Acanthamoeba cysts. A standard method for determining Acanthamoeba cyst viability in an in vitro cytotoxicity analysis is required to ensure that the double-walled and sturdy cysts are affected by the sub...

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Autores principales:	Nor Farah Azwani Che Mohamad, Nur Syakinah Nafisa Failei, Nurhidayana Mohd Rased, Azila Adnan, Ma Nyuk Ling, Hazlina Ahamad Zakeri, Eny Kusrini, Fatimah Hashim
Formato:	article
Lenguaje:	EN
Publicado:	Universitas Indonesia 2021
Materias:	dithiothreitol fluorescence intensity high content screening keratitis propidium iodide Technology T Technology (General) T1-995
Acceso en línea:	https://doaj.org/article/d03f68f8d3e045cab3bd4686ed4a5c67
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spelling	oai:doaj.org-article:d03f68f8d3e045cab3bd4686ed4a5c672021-12-02T17:41:43ZTechnique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity2086-96142087-210010.14716/ijtech.v12i4.4207https://doaj.org/article/d03f68f8d3e045cab3bd4686ed4a5c672021-10-01T00:00:00Zhttps://ijtech.eng.ui.ac.id/article/view/4207https://doaj.org/toc/2086-9614https://doaj.org/toc/2087-2100This study presents a straightforward and reliable method for determining the viability of Acanthamoeba cysts. A standard method for determining Acanthamoeba cyst viability in an in vitro cytotoxicity analysis is required to ensure that the double-walled and sturdy cysts are affected by the substance tested. In this study, a new approach was used to determine the cysticidal potential of redox Cleland’s reagent, dithiothreitol (DTT), against Acanthamoeba cysts. This approach constitutes a significant breakthrough, as the cyst form of Acanthamoeba is known for its high resistance to various chemicals and drugs used to treat infections of the central nervous system and eyes caused by Acanthamoeba. Cyst viability was evaluated based on the intensity of the cyst population under fluorescence produced by propidium iodide (PI) dye and measured using an enzyme-linked immunosorbent assay (ELISA) reader at an absorbance of 636 nm. The results were validated using high-content screening (HCS). For analysis, an individual cell was imaged and examined for phenotypic changes in the Acanthamoeba cyst at the cyst population level. Fluorescence intensity of the cysts in each well in a 96-well plate was measured using Image J software. HCS is an automated technique that uses fluorescence microscopy to produce quantitative data.Nor Farah Azwani Che MohamadNur Syakinah Nafisa FaileiNurhidayana Mohd RasedAzila AdnanMa Nyuk LingHazlina Ahamad ZakeriEny KusriniFatimah HashimUniversitas Indonesiaarticledithiothreitolfluorescence intensityhigh content screeningkeratitispropidium iodideTechnologyTTechnology (General)T1-995ENInternational Journal of Technology, Vol 12, Iss 4, Pp 720-727 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	dithiothreitol fluorescence intensity high content screening keratitis propidium iodide Technology T Technology (General) T1-995
spellingShingle	dithiothreitol fluorescence intensity high content screening keratitis propidium iodide Technology T Technology (General) T1-995 Nor Farah Azwani Che Mohamad Nur Syakinah Nafisa Failei Nurhidayana Mohd Rased Azila Adnan Ma Nyuk Ling Hazlina Ahamad Zakeri Eny Kusrini Fatimah Hashim Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
description	This study presents a straightforward and reliable method for determining the viability of Acanthamoeba cysts. A standard method for determining Acanthamoeba cyst viability in an in vitro cytotoxicity analysis is required to ensure that the double-walled and sturdy cysts are affected by the substance tested. In this study, a new approach was used to determine the cysticidal potential of redox Cleland’s reagent, dithiothreitol (DTT), against Acanthamoeba cysts. This approach constitutes a significant breakthrough, as the cyst form of Acanthamoeba is known for its high resistance to various chemicals and drugs used to treat infections of the central nervous system and eyes caused by Acanthamoeba. Cyst viability was evaluated based on the intensity of the cyst population under fluorescence produced by propidium iodide (PI) dye and measured using an enzyme-linked immunosorbent assay (ELISA) reader at an absorbance of 636 nm. The results were validated using high-content screening (HCS). For analysis, an individual cell was imaged and examined for phenotypic changes in the Acanthamoeba cyst at the cyst population level. Fluorescence intensity of the cysts in each well in a 96-well plate was measured using Image J software. HCS is an automated technique that uses fluorescence microscopy to produce quantitative data.
format	article
author	Nor Farah Azwani Che Mohamad Nur Syakinah Nafisa Failei Nurhidayana Mohd Rased Azila Adnan Ma Nyuk Ling Hazlina Ahamad Zakeri Eny Kusrini Fatimah Hashim
author_facet	Nor Farah Azwani Che Mohamad Nur Syakinah Nafisa Failei Nurhidayana Mohd Rased Azila Adnan Ma Nyuk Ling Hazlina Ahamad Zakeri Eny Kusrini Fatimah Hashim
author_sort	Nor Farah Azwani Che Mohamad
title	Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
title_short	Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
title_full	Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
title_fullStr	Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
title_full_unstemmed	Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity
title_sort	technique for determining the viability of acanthamoeba cysts treated with a cysticidal agent based on membrane integrity
publisher	Universitas Indonesia
publishDate	2021
url	https://doaj.org/article/d03f68f8d3e045cab3bd4686ed4a5c67
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Technique for Determining the Viability of Acanthamoeba Cysts Treated with a Cysticidal Agent Based on Membrane Integrity

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