Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors

Abstract Hexaminolevulinate (HAL) induced Protoporphyrin IX (PpIX) fluorescence is commonly used to differentiate cancer cells from normal cells in vivo, as for instance in blue light cystoscopy for bladder cancer diagnosis. A detailed approach is here provided to use this diagnostic principle ex vi...

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Autores principales:	Kit Man Chan, Jonathan Gleadle, Jordan Li, Thomas Danny Michl, Krasimir Vasilev, Melanie MacGregor
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9e6fea34737143f7aaefb15e4398d5c0
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spelling	oai:doaj.org-article:9e6fea34737143f7aaefb15e4398d5c02021-12-02T18:17:41ZImproving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors10.1038/s41598-021-86649-62045-2322https://doaj.org/article/9e6fea34737143f7aaefb15e4398d5c02021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86649-6https://doaj.org/toc/2045-2322Abstract Hexaminolevulinate (HAL) induced Protoporphyrin IX (PpIX) fluorescence is commonly used to differentiate cancer cells from normal cells in vivo, as for instance in blue light cystoscopy for bladder cancer diagnosis. A detailed approach is here provided to use this diagnostic principle ex vivo in an immunosensor device, towards enabling non-invasive cancer diagnostic from body fluids, such as urine. Several factors susceptible to affect the applicability of HAL-assisted diagnosis in body fluids were tested. These included the cell viability and its impact on PpIX fluorescence, the storage condition and shelf life of HAL premix reagent, light exposure (360–450 nm wavelengths) and its corresponding effect on both intensity and bleaching of the PpIX fluorescence as a function of the microscopy imaging conditions. There was no significant decrease in the viability of bladder cancer cells after 6 h at 4 °C (student’s t-test: p > 0.05). The cellular PpIX fluorescence decreased in a time-dependent manner when cancer cells were kept at 4 °C for extended period of time, though this didn’t significantly reduce the fluorescence intensity contrast between cancer and non-cancer cells kept in the same condition for 6 h. HAL premix reagent kept in long term storage at 4 °C induced stronger PpIX fluorescence than reagent kept in the − 20 °C freezer. The PpIX fluorescence was negatively affected by repeated light exposure but increased with illumination intensity and exposure time. Though this applied to both healthy and cancer cell lines, and therefore did not statistically improved the differentiation between cell types. This study revealed important experimental settings that need to be carefully considered to benefit from the analytical potential of HAL induced fluorescence when used in technologies for the diagnosis of cancer from body fluids.Kit Man ChanJonathan GleadleJordan LiThomas Danny MichlKrasimir VasilevMelanie MacGregorNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kit Man Chan Jonathan Gleadle Jordan Li Thomas Danny Michl Krasimir Vasilev Melanie MacGregor Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
description	Abstract Hexaminolevulinate (HAL) induced Protoporphyrin IX (PpIX) fluorescence is commonly used to differentiate cancer cells from normal cells in vivo, as for instance in blue light cystoscopy for bladder cancer diagnosis. A detailed approach is here provided to use this diagnostic principle ex vivo in an immunosensor device, towards enabling non-invasive cancer diagnostic from body fluids, such as urine. Several factors susceptible to affect the applicability of HAL-assisted diagnosis in body fluids were tested. These included the cell viability and its impact on PpIX fluorescence, the storage condition and shelf life of HAL premix reagent, light exposure (360–450 nm wavelengths) and its corresponding effect on both intensity and bleaching of the PpIX fluorescence as a function of the microscopy imaging conditions. There was no significant decrease in the viability of bladder cancer cells after 6 h at 4 °C (student’s t-test: p > 0.05). The cellular PpIX fluorescence decreased in a time-dependent manner when cancer cells were kept at 4 °C for extended period of time, though this didn’t significantly reduce the fluorescence intensity contrast between cancer and non-cancer cells kept in the same condition for 6 h. HAL premix reagent kept in long term storage at 4 °C induced stronger PpIX fluorescence than reagent kept in the − 20 °C freezer. The PpIX fluorescence was negatively affected by repeated light exposure but increased with illumination intensity and exposure time. Though this applied to both healthy and cancer cell lines, and therefore did not statistically improved the differentiation between cell types. This study revealed important experimental settings that need to be carefully considered to benefit from the analytical potential of HAL induced fluorescence when used in technologies for the diagnosis of cancer from body fluids.
format	article
author	Kit Man Chan Jonathan Gleadle Jordan Li Thomas Danny Michl Krasimir Vasilev Melanie MacGregor
author_facet	Kit Man Chan Jonathan Gleadle Jordan Li Thomas Danny Michl Krasimir Vasilev Melanie MacGregor
author_sort	Kit Man Chan
title	Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
title_short	Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
title_full	Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
title_fullStr	Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
title_full_unstemmed	Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
title_sort	improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9e6fea34737143f7aaefb15e4398d5c0
work_keys_str_mv	AT kitmanchan improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors AT jonathangleadle improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors AT jordanli improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors AT thomasdannymichl improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors AT krasimirvasilev improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors AT melaniemacgregor improvinghexaminolevulinateenabledcancercelldetectioninliquidbiopsyimmunosensors
_version_	1718378290280923136

Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors

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