In vitro optoacoustic flow cytometry with light scattering referencing

Abstract Morphological and functional optoacoustic imaging is enhanced by dedicated transgene reporters, in analogy to fluorescence methods. The development of optoacoustic reporters using protein engineering and directed evolution would be accelerated by high-throughput in-flow screening for intrac...

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Autores principales:	Markus Seeger, Andre C. Stiel, Vasilis Ntziachristos
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/071aebc2c9a34c0aa1647ad100e2686b
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spelling	oai:doaj.org-article:071aebc2c9a34c0aa1647ad100e2686b2021-12-02T14:16:07ZIn vitro optoacoustic flow cytometry with light scattering referencing10.1038/s41598-021-81584-y2045-2322https://doaj.org/article/071aebc2c9a34c0aa1647ad100e2686b2021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81584-yhttps://doaj.org/toc/2045-2322Abstract Morphological and functional optoacoustic imaging is enhanced by dedicated transgene reporters, in analogy to fluorescence methods. The development of optoacoustic reporters using protein engineering and directed evolution would be accelerated by high-throughput in-flow screening for intracellular, genetically encoded, optoacoustic contrast. However, accurate characterization of such contrast is impeded because the optoacoustic signals depend on the cell’s size and position in the flow chamber. We report herein an optoacoustic flow cytometer (OA-FCM) capable of precise measurement of intracellular optoacoustic signals of genetically-encoded chromoproteins in flow. The novel system records light-scattering as a reference for the detected optoacoustic signals in order to account for cell size and position, as well as excitation light flux in the focal volume, which we use to reference the detected optoacoustic signals to enhance the system’s precision. The OA-FCM was calibrated using micrometer-sized particles to showcase the ability to assess in-flow objects in the size range of single-cells. We demonstrate the capabilities of our OA-FCM to identify sub-populations in a mixture of two E. coli stocks expressing different reporter-proteins with a precision of over 90%. High-throughput screening of optoacoustic labels could pave the way for identifying genetically encoded optoacoustic reporters by transferring working concepts of the fluorescence field such as directed evolution and activated cell sorting.Markus SeegerAndre C. StielVasilis NtziachristosNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Markus Seeger Andre C. Stiel Vasilis Ntziachristos In vitro optoacoustic flow cytometry with light scattering referencing
description	Abstract Morphological and functional optoacoustic imaging is enhanced by dedicated transgene reporters, in analogy to fluorescence methods. The development of optoacoustic reporters using protein engineering and directed evolution would be accelerated by high-throughput in-flow screening for intracellular, genetically encoded, optoacoustic contrast. However, accurate characterization of such contrast is impeded because the optoacoustic signals depend on the cell’s size and position in the flow chamber. We report herein an optoacoustic flow cytometer (OA-FCM) capable of precise measurement of intracellular optoacoustic signals of genetically-encoded chromoproteins in flow. The novel system records light-scattering as a reference for the detected optoacoustic signals in order to account for cell size and position, as well as excitation light flux in the focal volume, which we use to reference the detected optoacoustic signals to enhance the system’s precision. The OA-FCM was calibrated using micrometer-sized particles to showcase the ability to assess in-flow objects in the size range of single-cells. We demonstrate the capabilities of our OA-FCM to identify sub-populations in a mixture of two E. coli stocks expressing different reporter-proteins with a precision of over 90%. High-throughput screening of optoacoustic labels could pave the way for identifying genetically encoded optoacoustic reporters by transferring working concepts of the fluorescence field such as directed evolution and activated cell sorting.
format	article
author	Markus Seeger Andre C. Stiel Vasilis Ntziachristos
author_facet	Markus Seeger Andre C. Stiel Vasilis Ntziachristos
author_sort	Markus Seeger
title	In vitro optoacoustic flow cytometry with light scattering referencing
title_short	In vitro optoacoustic flow cytometry with light scattering referencing
title_full	In vitro optoacoustic flow cytometry with light scattering referencing
title_fullStr	In vitro optoacoustic flow cytometry with light scattering referencing
title_full_unstemmed	In vitro optoacoustic flow cytometry with light scattering referencing
title_sort	in vitro optoacoustic flow cytometry with light scattering referencing
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/071aebc2c9a34c0aa1647ad100e2686b
work_keys_str_mv	AT markusseeger invitrooptoacousticflowcytometrywithlightscatteringreferencing AT andrecstiel invitrooptoacousticflowcytometrywithlightscatteringreferencing AT vasilisntziachristos invitrooptoacousticflowcytometrywithlightscatteringreferencing
_version_	1718391705843007488

In vitro optoacoustic flow cytometry with light scattering referencing

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