Dynamic Visualization and Quantification of Single Vesicle Opening and Content by Coupling Vesicle Impact Electrochemical Cytometry with Confocal Microscopy.

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Zheng, Ying-NingGothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology (author)

Dynamic Visualization and Quantification of Single Vesicle Opening and Content by Coupling Vesicle Impact Electrochemical Cytometry with Confocal Microscopy.

Article/chapterEnglish2021

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2021-08-09
American Chemical Society (ACS),2021

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LIBRIS-ID:oai:gup.ub.gu.se/313175
https://gup.ub.gu.se/publication/313175URI
https://doi.org/10.1021/acsmeasuresciau.1c00021DOI

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Language:English

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Subject category:ref swepub-contenttype
Subject category:art swepub-publicationtype

Notes

In this work, we introduce a novel method for visualization and quantitative measurement of the vesicle opening process by correlation of vesicle impact electrochemical cytometry (VIEC) with confocal microscopy. We have used a fluorophore conjugated to lipids to label the vesicle membrane and manipulate the membrane properties, which appears to make the membrane more susceptible to electroporation. The neurotransmitters inside the vesicles were visualized by use of a fluorescence false neurotransmitter 511 (FFN 511) through accumulation inside the vesicle via the neuronal vesicular monoamine transporter 2 (VMAT 2). Optical and electrochemical measurements of single vesicle electroporation were carried out using an in-house, disk-shaped, gold-modified ITO (Au/ITO) microelectrode device (5 nm thick, 33 μm diameter), which simultaneously acted as an electrode surface for VIEC and an optically transparent surface for confocal microscopy. As a result, the processes of adsorption, electroporation, and opening of single vesicles followed by neurotransmitter release on the Au/ITO surface have been simultaneously visualized and measured. Three opening patterns of single isolated vesicles were frequently observed. Comparing the vesicle opening patterns with their corresponding VIEC spikes, we propose that the behavior of the vesicular membrane on the electrode surface, including the adsorption time, residence time before vesicle opening, and the retention time after vesicle opening, are closely related to the vesicle content and size. Large vesicles with high content tend to adsorb to the electrode faster with higher frequency, followed by a shorter residence time before releasing their content, and their membrane remains on the electrode surface longer compared to the small vesicles with low content. With this approach, we start to unravel the vesicle opening process and to examine the fundamentals of exocytosis, supporting the proposed mechanism of partial or subquantal release in exocytosis.

Subject headings and genre

NATURVETENSKAP Kemi Analytisk kemi hsv//swe
NATURAL SCIENCES Chemical Sciences Analytical Chemistry hsv//eng
NATURVETENSKAP Kemi hsv//swe
NATURAL SCIENCES Chemical Sciences hsv//eng
Electrochemical Cytometry
electrochemistry
Confocal Microscopy
single vesicle imaging
dynamic
live imaging

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Nguyen, Tho D. K.Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xngduc (author)
Dunevall, JohanGothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xdunej (author)
Phan, Nhu TN,1981Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,National Center for Imaging Mass Spectrometry,Department of Chemistry and Molecular Biology(Swepub:gu)xphant (author)
Ewing, Andrew G,1957Gothenburg University,Göteborgs universitet,National Center for Imaging Mass Spectrometry,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xewian (author)
Göteborgs universitetInstitutionen för kemi och molekylärbiologi (creator_code:org_t)

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In:ACS measurement science Au: American Chemical Society (ACS)1:3, s. 131-1382694-250X

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By the author/editor: Zheng, Ying-Ning; Nguyen, Tho D. K ...; Dunevall, Johan; Phan, Nhu TN, 19 ...; Ewing, Andrew G, ...

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