Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.

Multicolor fluorescence microscopy is a powerful technique to fully visualize many biological phenomena by acquiring images from different spectrum channels. This study expands the scope of multicolor fluorescence microscopy by serial imaging of polystyrene micro-beads as surrogates for drug carrier...

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Autores principales:	Mert Kaya, Fabian Stein, Jeroen Rouwkema, Islam S M Khalil, Sarthak Misra
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ea4135b7ec3142589c22bd6357b54545
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spelling	oai:doaj.org-article:ea4135b7ec3142589c22bd6357b545452021-12-02T20:10:39ZSerial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.1932-620310.1371/journal.pone.0253222https://doaj.org/article/ea4135b7ec3142589c22bd6357b545452021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0253222https://doaj.org/toc/1932-6203Multicolor fluorescence microscopy is a powerful technique to fully visualize many biological phenomena by acquiring images from different spectrum channels. This study expands the scope of multicolor fluorescence microscopy by serial imaging of polystyrene micro-beads as surrogates for drug carriers, cancer spheroids formed using HeLa cells, and microfluidic channels. Three fluorophores with different spectral characteristics are utilized to perform multicolor microscopy. According to the spectrum analysis of the fluorophores, a multicolor widefield fluorescence microscope is developed. Spectral crosstalk is corrected by exciting the fluorophores in a round-robin manner and synchronous emitted light collection. To report the performance of the multicolor microscopy, a simplified 3D tumor model is created by placing beads and spheroids inside a channel filled with the cell culture medium is imaged at varying exposure times. As a representative case and a method for bio-hybrid drug carrier fabrication, a spheroid surface is coated with beads in a channel utilizing electrostatic forces under the guidance of multicolor microscopy. Our experiments show that multicolor fluorescence microscopy enables crosstalk-free and spectrally-different individual image acquisition of beads, spheroids, and channels with the minimum exposure time of 5.5 ms. The imaging technique has the potential to monitor drug carrier transportation to cancer cells in real-time.Mert KayaFabian SteinJeroen RouwkemaIslam S M KhalilSarthak MisraPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 6, p e0253222 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mert Kaya Fabian Stein Jeroen Rouwkema Islam S M Khalil Sarthak Misra Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
description	Multicolor fluorescence microscopy is a powerful technique to fully visualize many biological phenomena by acquiring images from different spectrum channels. This study expands the scope of multicolor fluorescence microscopy by serial imaging of polystyrene micro-beads as surrogates for drug carriers, cancer spheroids formed using HeLa cells, and microfluidic channels. Three fluorophores with different spectral characteristics are utilized to perform multicolor microscopy. According to the spectrum analysis of the fluorophores, a multicolor widefield fluorescence microscope is developed. Spectral crosstalk is corrected by exciting the fluorophores in a round-robin manner and synchronous emitted light collection. To report the performance of the multicolor microscopy, a simplified 3D tumor model is created by placing beads and spheroids inside a channel filled with the cell culture medium is imaged at varying exposure times. As a representative case and a method for bio-hybrid drug carrier fabrication, a spheroid surface is coated with beads in a channel utilizing electrostatic forces under the guidance of multicolor microscopy. Our experiments show that multicolor fluorescence microscopy enables crosstalk-free and spectrally-different individual image acquisition of beads, spheroids, and channels with the minimum exposure time of 5.5 ms. The imaging technique has the potential to monitor drug carrier transportation to cancer cells in real-time.
format	article
author	Mert Kaya Fabian Stein Jeroen Rouwkema Islam S M Khalil Sarthak Misra
author_facet	Mert Kaya Fabian Stein Jeroen Rouwkema Islam S M Khalil Sarthak Misra
author_sort	Mert Kaya
title	Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
title_short	Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
title_full	Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
title_fullStr	Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
title_full_unstemmed	Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
title_sort	serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/ea4135b7ec3142589c22bd6357b54545
work_keys_str_mv	AT mertkaya serialimagingofmicroagentsandcancercellspheroidsinamicrofluidicchannelusingmulticolorfluorescencemicroscopy AT fabianstein serialimagingofmicroagentsandcancercellspheroidsinamicrofluidicchannelusingmulticolorfluorescencemicroscopy AT jeroenrouwkema serialimagingofmicroagentsandcancercellspheroidsinamicrofluidicchannelusingmulticolorfluorescencemicroscopy AT islamsmkhalil serialimagingofmicroagentsandcancercellspheroidsinamicrofluidicchannelusingmulticolorfluorescencemicroscopy AT sarthakmisra serialimagingofmicroagentsandcancercellspheroidsinamicrofluidicchannelusingmulticolorfluorescencemicroscopy
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Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.

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