Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy

Ralf P Friedrich,1 Eveline Schreiber,1 Rainer Tietze,1 Hai Yang,2 Christian Pilarsky,2 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung-Professorship, Universitätsk...

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Autores principales:	Friedrich RP, Schreiber E, Tietze R, Yang H, Pilarsky C, Alexiou C
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	superparamagnetic iron oxide nanoparticles cellular spion uptake cytotoxicity flow cytometry atomic emission spectroscopy Medical technology R855-855.5 Chemical technology TP1-1185
Acceso en línea:	https://doaj.org/article/55444c097945491f8694d51481e70f30
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spelling	oai:doaj.org-article:55444c097945491f8694d51481e70f302021-12-02T16:10:55ZIntracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy1177-8903https://doaj.org/article/55444c097945491f8694d51481e70f302020-12-01T00:00:00Zhttps://www.dovepress.com/intracellular-quantification-and-localization-of-label-free-iron-oxide-peer-reviewed-article-NSAhttps://doaj.org/toc/1177-8903Ralf P Friedrich,1 Eveline Schreiber,1 Rainer Tietze,1 Hai Yang,2 Christian Pilarsky,2 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung-Professorship, Universitätsklinikum Erlangen, Erlangen, 91054, Germany; 2Department of Surgery, Universitätsklinikum Erlangen, Erlangen 91054, GermanyCorrespondence: Ralf P FriedrichDepartment of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung-Professorship, Universitätsklinikum Erlangen, Glückstrasse 10a, Erlangen 91054, GermanyTel +49 9131 85 43943Fax +49 9131 85 34828Email ralf.friedrich@uk-erlangen.deBackground: The limitations of optical microscopy to determine the cellular localization of label-free nanoparticles prevent a solid prediction of the cellular effect of particles intended for medical applications. To avoid the strong physicochemical changes associated with fluorescent labelling, which often result in differences in cellular uptake, efficiency and toxicity of particles, novel detection techniques are required.Methods: In the present study, we determined the intracellular content of unlabeled SPIONs by analyzing refractive index (RI)-based images from holotomographic three-dimensional (3D) microscopy and side scatter data measured by flow cytometry. The results were compared with the actual cellular SPION amount as quantified by atomic emission spectroscopy (AES).Results: Live cell imaging by 3D holotomographic microscopy demonstrated cell-specific differences in intracellular nanoparticle uptake in different pancreatic cell lines. Thus, treatment of PANC-1SMAD4 (1− 4) and PANC-1SMAD4 (2− 6) with SPIONs resulted in a significant increase in number of areas with higher RI, whereas in PANC-1, SUIT-2 and PaCa DD183, only a minimal increase of spots with high RI was observed. The increase in areas with high RI was in accordance with the SPION content determined by quantitative iron measurements using AES. In contrast, determination of the SPION amount by flow cytometry was strongly cell type-dependent and did not allow the discrimination between intracellular and membrane-bound SPIONs. However, flow cytometry is a very rapid and reliable method to assess the cellular toxicity and allows an estimation of the cell-associated SPION content.Conclusion: Holotomographic 3D microscopy is a useful method to distinguish between intracellular and membrane-associated particles. Thus, it provides a valuable tool for scientists to evaluate the cellular localization and the particle load, which facilitates prediction of potential toxicity and efficiency of nanoparticles for medical applications.Keywords: superparamagnetic iron oxide nanoparticles, cellular SPION uptake, cytotoxicity, flow cytometry, atomic emission spectroscopyFriedrich RPSchreiber ETietze RYang HPilarsky CAlexiou CDove Medical Pressarticlesuperparamagnetic iron oxide nanoparticlescellular spion uptakecytotoxicityflow cytometryatomic emission spectroscopyMedical technologyR855-855.5Chemical technologyTP1-1185ENNanotechnology, Science and Applications, Vol Volume 13, Pp 119-130 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	superparamagnetic iron oxide nanoparticles cellular spion uptake cytotoxicity flow cytometry atomic emission spectroscopy Medical technology R855-855.5 Chemical technology TP1-1185
spellingShingle	superparamagnetic iron oxide nanoparticles cellular spion uptake cytotoxicity flow cytometry atomic emission spectroscopy Medical technology R855-855.5 Chemical technology TP1-1185 Friedrich RP Schreiber E Tietze R Yang H Pilarsky C Alexiou C Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
description	Ralf P Friedrich,1 Eveline Schreiber,1 Rainer Tietze,1 Hai Yang,2 Christian Pilarsky,2 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung-Professorship, Universitätsklinikum Erlangen, Erlangen, 91054, Germany; 2Department of Surgery, Universitätsklinikum Erlangen, Erlangen 91054, GermanyCorrespondence: Ralf P FriedrichDepartment of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung-Professorship, Universitätsklinikum Erlangen, Glückstrasse 10a, Erlangen 91054, GermanyTel +49 9131 85 43943Fax +49 9131 85 34828Email ralf.friedrich@uk-erlangen.deBackground: The limitations of optical microscopy to determine the cellular localization of label-free nanoparticles prevent a solid prediction of the cellular effect of particles intended for medical applications. To avoid the strong physicochemical changes associated with fluorescent labelling, which often result in differences in cellular uptake, efficiency and toxicity of particles, novel detection techniques are required.Methods: In the present study, we determined the intracellular content of unlabeled SPIONs by analyzing refractive index (RI)-based images from holotomographic three-dimensional (3D) microscopy and side scatter data measured by flow cytometry. The results were compared with the actual cellular SPION amount as quantified by atomic emission spectroscopy (AES).Results: Live cell imaging by 3D holotomographic microscopy demonstrated cell-specific differences in intracellular nanoparticle uptake in different pancreatic cell lines. Thus, treatment of PANC-1SMAD4 (1− 4) and PANC-1SMAD4 (2− 6) with SPIONs resulted in a significant increase in number of areas with higher RI, whereas in PANC-1, SUIT-2 and PaCa DD183, only a minimal increase of spots with high RI was observed. The increase in areas with high RI was in accordance with the SPION content determined by quantitative iron measurements using AES. In contrast, determination of the SPION amount by flow cytometry was strongly cell type-dependent and did not allow the discrimination between intracellular and membrane-bound SPIONs. However, flow cytometry is a very rapid and reliable method to assess the cellular toxicity and allows an estimation of the cell-associated SPION content.Conclusion: Holotomographic 3D microscopy is a useful method to distinguish between intracellular and membrane-associated particles. Thus, it provides a valuable tool for scientists to evaluate the cellular localization and the particle load, which facilitates prediction of potential toxicity and efficiency of nanoparticles for medical applications.Keywords: superparamagnetic iron oxide nanoparticles, cellular SPION uptake, cytotoxicity, flow cytometry, atomic emission spectroscopy
format	article
author	Friedrich RP Schreiber E Tietze R Yang H Pilarsky C Alexiou C
author_facet	Friedrich RP Schreiber E Tietze R Yang H Pilarsky C Alexiou C
author_sort	Friedrich RP
title	Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
title_short	Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
title_full	Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
title_fullStr	Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
title_full_unstemmed	Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy
title_sort	intracellular quantification and localization of label-free iron oxide nanoparticles by holotomographic microscopy
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/55444c097945491f8694d51481e70f30
work_keys_str_mv	AT friedrichrp intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy AT schreibere intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy AT tietzer intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy AT yangh intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy AT pilarskyc intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy AT alexiouc intracellularquantificationandlocalizationoflabelfreeironoxidenanoparticlesbyholotomographicmicroscopy
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Intracellular Quantification and Localization of Label-Free Iron Oxide Nanoparticles by Holotomographic Microscopy

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