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用于多色荧光激活细胞分选的稳健双乳液。

Robust Double Emulsions for Multicolor Fluorescence-Activated Cell Sorting.

机构信息

Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zürich, 8093 Zürich, Switzerland.

Institute for Research in Biomedicine, Faculty of Biomedical Sciences, Università della Svizzera italiana, 6500 Bellinzona, Switzerland.

出版信息

Anal Chem. 2024 Sep 17;96(37):14809-14818. doi: 10.1021/acs.analchem.4c02363. Epub 2024 Sep 4.


DOI:10.1021/acs.analchem.4c02363
PMID:39231502
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11411495/
Abstract

Cell-cell interactions are essential for the proper functioning of multicellular organisms. For example, T cells interact with antigen-presenting cells (APCs) through specific T-cell receptor (TCR)-antigen interactions during an immune response. Fluorescence-activated droplet sorting (FADS) is a high-throughput technique for efficiently screening cellular interaction events. Unfortunately, current droplet sorting instruments have significant limitations, most notably related to analytical throughput and complex operation. In contrast, commercial fluorescence-activated cell sorters offer superior speed, sensitivity, and multiplexing capabilities, although their use as droplet sorters is poorly defined and underutilized. Herein, we present a universally applicable and simple-to-implement workflow for generating double emulsions and performing multicolor cell sorting using a commercial FACS instrument. This workflow achieves a double emulsion detection rate exceeding 90%, enabling multicellular encapsulation and high-throughput immune cell activation sorting for the first time. We anticipate that the presented droplet sorting strategy will benefit cell biology laboratories by providing access to an advanced microfluidic toolbox with minimal effort and cost investment.

摘要

细胞间相互作用对于多细胞生物的正常功能至关重要。例如,在免疫反应中,T 细胞通过特异性 T 细胞受体(TCR)-抗原相互作用与抗原呈递细胞(APC)相互作用。荧光激活液滴分选(FADS)是一种高效筛选细胞相互作用事件的高通量技术。不幸的是,目前的液滴分选仪器具有显著的局限性,最明显的是与分析吞吐量和复杂操作有关。相比之下,商用荧光激活细胞分选器具有卓越的速度、灵敏度和多重检测能力,尽管它们作为液滴分选器的应用尚未得到明确界定和充分利用。在此,我们提出了一种普遍适用且易于实施的工作流程,用于使用商用 FACS 仪器生成双乳液并进行多色细胞分选。该工作流程实现了超过 90%的双乳液检测率,首次实现了多细胞封装和高通量免疫细胞激活分选。我们预计,所提出的液滴分选策略将通过提供先进的微流控工具包,使细胞生物学实验室能够以最小的努力和成本投入获得该工具包,从而使细胞生物学实验室受益。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/f134837c9661/ac4c02363_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/3b8ccdee8cfa/ac4c02363_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/e69a21002006/ac4c02363_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/de94c28f6216/ac4c02363_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/f134837c9661/ac4c02363_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/3b8ccdee8cfa/ac4c02363_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/e69a21002006/ac4c02363_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/de94c28f6216/ac4c02363_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01f3/11411495/f134837c9661/ac4c02363_0004.jpg

相似文献

[1]
Robust Double Emulsions for Multicolor Fluorescence-Activated Cell Sorting.

Anal Chem. 2024-9-17

[2]
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[3]
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[4]
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[5]
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Cytometry A. 2024-7

[6]
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[7]
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[8]
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[9]
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[10]
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引用本文的文献

[1]
Deciphering cellular complexity: advances and future directions in single-cell protein analysis.

Front Bioeng Biotechnol. 2025-1-14

本文引用的文献

[1]
Design automation of microfluidic single and double emulsion droplets with machine learning.

Nat Commun. 2024-1-2

[2]
Double emulsions as a high-throughput enrichment and isolation platform for slower-growing microbes.

ISME Commun. 2023-5-9

[3]
Combi-seq for multiplexed transcriptome-based profiling of drug combinations using deterministic barcoding in single-cell droplets.

Nat Commun. 2022-8-1

[4]
High-throughput single-cell antibody secretion quantification and enrichment using droplet microfluidics-based FRET assay.

iScience. 2022-6-2

[5]
Systematic characterization of effect of flow rates and buffer compositions on double emulsion droplet volumes and stability.

Lab Chip. 2022-6-14

[6]
An efficient strategy for a controllable droplet merging system for digital analysis.

RSC Adv. 2018-10-5

[7]
Droplet flow cytometry for single-cell analysis.

RSC Adv. 2021-6-14

[8]
Cell Culture in Microfluidic Droplets.

Chem Rev. 2022-4-13

[9]
Characterizing cell interactions at scale with made-to-order droplet ensembles (MODEs).

Proc Natl Acad Sci U S A. 2022-2-1

[10]
Immunotherapy of cancer in single-cell RNA sequencing era: A precision medicine perspective.

Biomed Pharmacother. 2022-2

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