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Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy

Composed of a fluidic and an optical system, flow cytometry has been widely used for biosensing. The fluidic flow enables its automatic high-throughput sample loading and sorting while the optical system works for molecular detection by fluorescence for micron-level cells and particles. This technol...

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Detalles Bibliográficos
Autores principales: Zhao, Xiaohui, Ding, Leqi, Yan, Jingsheng, Xu, Jin, He, Hao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10059927/
https://www.ncbi.nlm.nih.gov/pubmed/36992015
http://dx.doi.org/10.3390/s23063305
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author Zhao, Xiaohui
Ding, Leqi
Yan, Jingsheng
Xu, Jin
He, Hao
author_facet Zhao, Xiaohui
Ding, Leqi
Yan, Jingsheng
Xu, Jin
He, Hao
author_sort Zhao, Xiaohui
collection PubMed
description Composed of a fluidic and an optical system, flow cytometry has been widely used for biosensing. The fluidic flow enables its automatic high-throughput sample loading and sorting while the optical system works for molecular detection by fluorescence for micron-level cells and particles. This technology is quite powerful and highly developed; however, it requires a sample in the form of a suspension and thus only works in vitro. In this study, we report a simple scheme to construct a flow cytometry based on a confocal microscope without any modifications. We demonstrate that line scanning of microscopy can effectively excite fluorescence of flowing microbeads or cells in a capillary tube in vitro and in blood vessels of live mice in vivo. This method can resolve microbeads at several microns and the results are comparable to a classic flow cytometer. The absolute diameter of flowing samples can be indicated directly. The sampling limitations and variations of this method is carefully analyzed. This scheme can be easily accomplished by any commercial confocal microscope systems, expands the function of them, and is of promising potential for simultaneous confocal microscopy and in vivo detection of cells in blood vessels of live animals by a single system.
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spelling pubmed-100599272023-03-30 Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy Zhao, Xiaohui Ding, Leqi Yan, Jingsheng Xu, Jin He, Hao Sensors (Basel) Communication Composed of a fluidic and an optical system, flow cytometry has been widely used for biosensing. The fluidic flow enables its automatic high-throughput sample loading and sorting while the optical system works for molecular detection by fluorescence for micron-level cells and particles. This technology is quite powerful and highly developed; however, it requires a sample in the form of a suspension and thus only works in vitro. In this study, we report a simple scheme to construct a flow cytometry based on a confocal microscope without any modifications. We demonstrate that line scanning of microscopy can effectively excite fluorescence of flowing microbeads or cells in a capillary tube in vitro and in blood vessels of live mice in vivo. This method can resolve microbeads at several microns and the results are comparable to a classic flow cytometer. The absolute diameter of flowing samples can be indicated directly. The sampling limitations and variations of this method is carefully analyzed. This scheme can be easily accomplished by any commercial confocal microscope systems, expands the function of them, and is of promising potential for simultaneous confocal microscopy and in vivo detection of cells in blood vessels of live animals by a single system. MDPI 2023-03-21 /pmc/articles/PMC10059927/ /pubmed/36992015 http://dx.doi.org/10.3390/s23063305 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Zhao, Xiaohui
Ding, Leqi
Yan, Jingsheng
Xu, Jin
He, Hao
Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title_full Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title_fullStr Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title_full_unstemmed Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title_short Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy
title_sort constructing an in vitro and in vivo flow cytometry by fast line scanning of confocal microscopy
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10059927/
https://www.ncbi.nlm.nih.gov/pubmed/36992015
http://dx.doi.org/10.3390/s23063305
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