Please use this identifier to cite or link to this item:
http://bura.brunel.ac.uk/handle/2438/25426
Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Haas, KT | - |
dc.contributor.author | Fries, MW | - |
dc.contributor.author | Venkitaraman, AR | - |
dc.contributor.author | Esposito, A | - |
dc.date.accessioned | 2022-11-03T12:54:47Z | - |
dc.date.available | 2022-11-03T12:54:47Z | - |
dc.date.issued | 2021-05-27 | - |
dc.identifier | ORCiD ID: Alessandro Esposito: https://orcid.org/0000-0002-5051-091X | - |
dc.identifier | 637123 | - |
dc.identifier.citation | Haas, K.T. et al. (2021) 'Single-Cell Biochemical Multiplexing by Multidimensional Phasor Demixing and Spectral Fluorescence Lifetime Imaging Microscopy', Frontiers in Physics, 9, 637123, pp. 1 - 14. doi: 10.3389/fphy.2021.637123. | en_US |
dc.identifier.uri | https://bura.brunel.ac.uk/handle/2438/25426 | - |
dc.description | Data Availability Statement: The original contributions presented in the study are included in the article/Supplementary Material; further inquiries can be directed to the corresponding author. | en_US |
dc.description.abstract | Copyright © 2021 Haas, Fries, Venkitaraman and Esposito. Revealing mechanisms underpinning cell function requires understanding the relationship between different biochemical reactions in living cells. However, our capabilities to monitor more than two biochemical reactions in living cells are limited. Therefore, the development of methods for real-time biochemical multiplexing is of fundamental importance. Here, we show that data acquired with multicolor (mcFLIM) or spectrally resolved (sFLIM) fluorescence lifetime imaging can be conveniently described with multidimensional phasor transforms. We demonstrate a computational framework capable of demixing three Forster resonance energy transfer (FRET) probes and quantifying multiplexed biochemical activities in single living cells. We provide a comparison between mcFLIM and sFLIM suggesting that sFLIM might be advantageous for the future development of heavily multiplexed assays. However, mcFLIM—more readily available with commercial systems—can be applied for the concomitant monitoring of three enzymes in living cells without significant losses. | en_US |
dc.description.sponsorship | Gates Foundation studentship; Medical Research Council core grants (MC_UU_12022/1 and MC_UU_12022/8); Wellcome Trust (090340/Z/09/Z); Cancer Research UK (C54674/A27487). | en_US |
dc.format.extent | 1 - 14 | - |
dc.format.medium | Electronic | - |
dc.language | English | - |
dc.language.iso | en_US | en_US |
dc.publisher | Frontiers Media SA | en_US |
dc.rights | Copyright © 2021 Haas, Fries, Venkitaraman and Esposito. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. | - |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | - |
dc.subject | sFLIM | en_US |
dc.subject | FRET biosensors | en_US |
dc.subject | FRET biosensors | en_US |
dc.subject | TCSPC | en_US |
dc.subject | spectral demixing | en_US |
dc.subject | biochemical multiplexing | en_US |
dc.title | Single-Cell Biochemical Multiplexing by Multidimensional Phasor Demixing and Spectral Fluorescence Lifetime Imaging Microscopy | en_US |
dc.type | Article | en_US |
dc.identifier.doi | https://doi.org/10.3389/fphy.2021.637123 | - |
dc.relation.isPartOf | Frontiers in Physics | - |
pubs.publication-status | Published | - |
pubs.volume | 9 | - |
dc.identifier.eissn | 2296-424X | - |
dc.rights.holder | Haas, Fries, Venkitaraman and Esposito | - |
Appears in Collections: | Dept of Life Sciences Research Papers |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
FullText.pdf | Copyright © 2021 Haas, Fries, Venkitaraman and Esposito. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. | 2.96 MB | Adobe PDF | View/Open |
This item is licensed under a Creative Commons License