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Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer

Conventional flow cytometry is a valuable quantitative tool. Flow cytometers reveal physical and biochemical information from cells at a high throughput, which is quite valuable for many biomedical, biological, and diagnostic research fields. Flow cytometers range in complexity and typically provide...

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Autores principales: Sambrano, Jesus, Rodriguez, Felicia, Martin, John, Houston, Jessica P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8357029/
https://www.ncbi.nlm.nih.gov/pubmed/34386487
http://dx.doi.org/10.3389/fphy.2021.647985
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author Sambrano, Jesus
Rodriguez, Felicia
Martin, John
Houston, Jessica P.
author_facet Sambrano, Jesus
Rodriguez, Felicia
Martin, John
Houston, Jessica P.
author_sort Sambrano, Jesus
collection PubMed
description Conventional flow cytometry is a valuable quantitative tool. Flow cytometers reveal physical and biochemical information from cells at a high throughput, which is quite valuable for many biomedical, biological, and diagnostic research fields. Flow cytometers range in complexity and typically provide multiparametric data for the user at rates of up to 50,000 cells measured per second. Cytometry systems are configured such that fluorescence or scattered light signals are collected per-cell, and the integrated optical signal at a given wavelength range indicates a particular cellular feature such as phenotype or morphology. When the timing of the optical signal is measured, the cytometry system becomes “time-resolved.” Time-resolved flow cytometry (TRFC) instruments can detect fluorescence decay kinetics, and such measurements are consequential for Förster Resonance Energy Transfer (FRET) studies, multiplexing, and metabolic mapping, to name a few. TRFC systems capture fluorescence lifetimes at rates of thousands of cells per-second, however the approach is challenged at this throughput by terminal cellular velocities. High flow rates limit the total number of photons integrated per-cell, reducing the reliability of the average lifetime as a cytometric parameter. In this contribution, we examine an innovative approach to address this signal-to-noise issue. The technology merges time-resolved hardware with microfluidics and acoustics. We present an “acoustofluidic” time-resolved flow cytometer so that cellular velocities can be adjusted on the fly with a standing acoustic wave (SAW). Our work shows that acoustic control can be combined with time-resolved features to appropriately balance the throughput with the optical signals necessary for lifetime data.
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spelling pubmed-83570292021-08-11 Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer Sambrano, Jesus Rodriguez, Felicia Martin, John Houston, Jessica P. Front Phys Article Conventional flow cytometry is a valuable quantitative tool. Flow cytometers reveal physical and biochemical information from cells at a high throughput, which is quite valuable for many biomedical, biological, and diagnostic research fields. Flow cytometers range in complexity and typically provide multiparametric data for the user at rates of up to 50,000 cells measured per second. Cytometry systems are configured such that fluorescence or scattered light signals are collected per-cell, and the integrated optical signal at a given wavelength range indicates a particular cellular feature such as phenotype or morphology. When the timing of the optical signal is measured, the cytometry system becomes “time-resolved.” Time-resolved flow cytometry (TRFC) instruments can detect fluorescence decay kinetics, and such measurements are consequential for Förster Resonance Energy Transfer (FRET) studies, multiplexing, and metabolic mapping, to name a few. TRFC systems capture fluorescence lifetimes at rates of thousands of cells per-second, however the approach is challenged at this throughput by terminal cellular velocities. High flow rates limit the total number of photons integrated per-cell, reducing the reliability of the average lifetime as a cytometric parameter. In this contribution, we examine an innovative approach to address this signal-to-noise issue. The technology merges time-resolved hardware with microfluidics and acoustics. We present an “acoustofluidic” time-resolved flow cytometer so that cellular velocities can be adjusted on the fly with a standing acoustic wave (SAW). Our work shows that acoustic control can be combined with time-resolved features to appropriately balance the throughput with the optical signals necessary for lifetime data. 2021-05-14 2021-05 /pmc/articles/PMC8357029/ /pubmed/34386487 http://dx.doi.org/10.3389/fphy.2021.647985 Text en https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) (https://creativecommons.org/licenses/by/4.0/) . 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.
spellingShingle Article
Sambrano, Jesus
Rodriguez, Felicia
Martin, John
Houston, Jessica P.
Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title_full Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title_fullStr Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title_full_unstemmed Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title_short Toward the Development of an On-Chip Acoustic Focusing Fluorescence Lifetime Flow Cytometer
title_sort toward the development of an on-chip acoustic focusing fluorescence lifetime flow cytometer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8357029/
https://www.ncbi.nlm.nih.gov/pubmed/34386487
http://dx.doi.org/10.3389/fphy.2021.647985
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