Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms

Abstract Continuous monitoring of large specimens for long durations requires fast volume imaging. This is essential for understanding the processes occurring during the developmental stages of multicellular organisms. One of the key obstacles of fluorescence based prolonged monitoring and data coll...

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Autores principales:	Jigmi Basumatary, Tarannum Ara, Amartya Mukherjee, Debanjan Dutta, Upendra Nongthomba, Partha Pratim Mondal
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d9df9a8a9ccc4973a9a41c5c71c70777
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spelling	oai:doaj.org-article:d9df9a8a9ccc4973a9a41c5c71c707772021-12-02T14:17:31ZFluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms10.1038/s41598-021-86951-32045-2322https://doaj.org/article/d9df9a8a9ccc4973a9a41c5c71c707772021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86951-3https://doaj.org/toc/2045-2322Abstract Continuous monitoring of large specimens for long durations requires fast volume imaging. This is essential for understanding the processes occurring during the developmental stages of multicellular organisms. One of the key obstacles of fluorescence based prolonged monitoring and data collection is photobleaching. To capture the biological processes and simultaneously overcome the effect of bleaching, we developed single- and multi-color lightsheet based OVSS imaging technique that enables rapid screening of multiple tissues in an organism. Our approach based on OVSS imaging employs quantized step rotation of the specimen to record 2D angular data that reduces data acquisition time when compared to the existing light sheet imaging system (SPIM). A co-planar multicolor light sheet PSF is introduced to illuminate the tissues labelled with spectrally-separated fluorescent probes. The detection is carried out using a dual-channel sub-system that can simultaneously record spectrally separate volume stacks of the target organ. Arduino-based control systems were employed to automatize and control the volume data acquisition process. To illustrate the advantages of our approach, we have noninvasively imaged the Drosophila larvae and Zebrafish embryo. Dynamic studies of multiple organs (muscle and yolk-sac) in Zebrafish for a prolonged duration (5 days) were carried out to understand muscle structuring (Dystrophin, microfibers), primitive Macrophages (in yolk-sac) and inter-dependent lipid and protein-based metabolism. The volume-based study, intensity line-plots and inter-dependence ratio analysis allowed us to understand the transition from lipid-based metabolism to protein-based metabolism during early development (Pharyngula period with a critical transition time, $$\tau _c = 50$$ τ c = 50 h post-fertilization) in Zebrafish. The advantage of multicolor lightsheet illumination, fast volume scanning, simultaneous visualization of multiple organs and an order-less photobleaching makes OVSS imaging the system of choice for rapid monitoring and real-time assessment of macroscopic biological organisms with microscopic resolution.Jigmi BasumataryTarannum AraAmartya MukherjeeDebanjan DuttaUpendra NongthombaPartha Pratim MondalNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jigmi Basumatary Tarannum Ara Amartya Mukherjee Debanjan Dutta Upendra Nongthomba Partha Pratim Mondal Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
description	Abstract Continuous monitoring of large specimens for long durations requires fast volume imaging. This is essential for understanding the processes occurring during the developmental stages of multicellular organisms. One of the key obstacles of fluorescence based prolonged monitoring and data collection is photobleaching. To capture the biological processes and simultaneously overcome the effect of bleaching, we developed single- and multi-color lightsheet based OVSS imaging technique that enables rapid screening of multiple tissues in an organism. Our approach based on OVSS imaging employs quantized step rotation of the specimen to record 2D angular data that reduces data acquisition time when compared to the existing light sheet imaging system (SPIM). A co-planar multicolor light sheet PSF is introduced to illuminate the tissues labelled with spectrally-separated fluorescent probes. The detection is carried out using a dual-channel sub-system that can simultaneously record spectrally separate volume stacks of the target organ. Arduino-based control systems were employed to automatize and control the volume data acquisition process. To illustrate the advantages of our approach, we have noninvasively imaged the Drosophila larvae and Zebrafish embryo. Dynamic studies of multiple organs (muscle and yolk-sac) in Zebrafish for a prolonged duration (5 days) were carried out to understand muscle structuring (Dystrophin, microfibers), primitive Macrophages (in yolk-sac) and inter-dependent lipid and protein-based metabolism. The volume-based study, intensity line-plots and inter-dependence ratio analysis allowed us to understand the transition from lipid-based metabolism to protein-based metabolism during early development (Pharyngula period with a critical transition time, $$\tau _c = 50$$ τ c = 50 h post-fertilization) in Zebrafish. The advantage of multicolor lightsheet illumination, fast volume scanning, simultaneous visualization of multiple organs and an order-less photobleaching makes OVSS imaging the system of choice for rapid monitoring and real-time assessment of macroscopic biological organisms with microscopic resolution.
format	article
author	Jigmi Basumatary Tarannum Ara Amartya Mukherjee Debanjan Dutta Upendra Nongthomba Partha Pratim Mondal
author_facet	Jigmi Basumatary Tarannum Ara Amartya Mukherjee Debanjan Dutta Upendra Nongthomba Partha Pratim Mondal
author_sort	Jigmi Basumatary
title	Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
title_short	Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
title_full	Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
title_fullStr	Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
title_full_unstemmed	Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms
title_sort	fluorescence based rapid optical volume screening system (ovss) for interrogating multicellular organisms
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d9df9a8a9ccc4973a9a41c5c71c70777
work_keys_str_mv	AT jigmibasumatary fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms AT tarannumara fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms AT amartyamukherjee fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms AT debanjandutta fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms AT upendranongthomba fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms AT parthapratimmondal fluorescencebasedrapidopticalvolumescreeningsystemovssforinterrogatingmulticellularorganisms
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Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms

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