Linear triglycerol-based fluorosurfactants show high potential for droplet-microfluidics-based biochemical assays

Chowdhury, Mohammad Suman; Zheng, Wenshan; Singh, Abhishek Kumar; Ong, Irvine Lian Hao; Hou, Yong; Heyman, John A.; Faghani, Abbas; Amstad, Esther; Weitz, David A.; Haag, Rainer

doi:10.1039/d1sm00890k

Cited by 13 publications

(28 citation statements)

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“…Consequently, we decided to pursue the study with an average DOC of 5.0 and the tetra-azide fluorosurfactant (3) to limit the fraction of residual nongrafted antibody in the droplets. The size distribution of droplets was studied using a method previously published by Chowdhury et al, 25 and the droplet size distribution of the emulsion stabilized by the fluorosurfactant 3 showed a coefficient of variation of 5.1% after 2 h (Figure S2).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Droplet Surface Immunoassay by Relocation (D-SIRe) for High-Throughput Analysis of Cytosolic Proteins at the Single-Cell Level

et al. 2023

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Enzyme-linked immunosorbent assay (ELISA) is a central analytic method in biological science for the detection of proteins. Introduction of droplet-based microfluidics allowed the development of miniaturized, less-consuming, and more sensitive ELISA assays by coencapsulating the biological sample and antibody-functionalized particles. We report herein an alternative in-droplet immunoassay format, which avoids the use of particles. It exploits the oil/aqueous-phase interface as a protein capture and detection surface. This is achieved using tailored perfluorinated surfactants bearing azide-functionalized PEG-based polar headgroups, which spontaneously react when meeting at the droplet formation site, with strained alkyne-functionalized antibodies solubilized in the water phase. The resulting antibody-functionalized inner surface can then be used to capture a target protein. This surface capture process leads to concomitant relocation at the surface of a labeled detection antibody and in turn to a drastic change in the shape of the fluorescence signal from a convex shape (not captured) to a characteristic concave shape (captured). This novel droplet surface immunoassay by fluorescence relocation (D-SIRe) proved to be fast and sensitive at 2.3 attomoles of analyte per droplet. It was further demonstrated to allow detection of cytosolic proteins at the single bacteria level.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Droplet Surface Immunoassay by Relocation (D-SIRe) for High-Throughput Analysis of Cytosolic Proteins at the Single-Cell Level

et al. 2023

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“…Diese Eigenschaft hat spannende Forschungs-und Produktentwicklungen in Chemie und Biologie angestoßen, unter anderem organische Synthesen im kleinen Maßstab, Single-Cell-Barcoding, Einzelzell-Chromatin-Immunpräzipitationssequenzierung, kombinatorisches Wirkstoffscreening, Entdeckung therapeutischer Antikörper, gerichtete enzymatische Evolution und digitale enzymbasierte Immunadsorptionstests (ELISA). [13,20] Oft sind diese Tröpfchen metastabil und neigen zu vorzeitiger Koaleszenz, was mit Fluortensiden, die ihre Stabilität erhöhen, vermieden werden kann. Zum Lösen dieser Fluortenside wird in der Regel ein biologisch inertes fluoriertes Öl verwendet, das mit wässrigen und organischen Lösungen nicht mischbar ist und somit eine ideale Plattform für das Einfangen von Biomolekülen in Tröpfchen darstellt.…”

Section: Stabilisierung Von Mikrotröpfchenunclassified

Grenzflächen mit fluorierten Amphiphilen: Überstrukturen und Mikrofluidik

et al. 2023

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Abbildung 4. a) Allgemeine chemische Struktur der Gelatoren auf der Grundlage des Cyclohexyl-Diamid-Kerns. b) Vorgeschlagener Aufbaumechanismus für den C 7 F 15 -substituierten Gelator. [31] c)-e) Zweistufige hierarchische Struktur der selbstorganisierten Strukturen von Gelatoren mit verschiedenen Seitenketten. [31,32] f)-h) Selbstreinigung gegenüber Blut auf geschmierter gleitfähiger Oberfläche. [31] i) und j) Lichtmikroskopische Aufnahmen, die das unterschiedliche Ausmaß der Beeinträchtigung während der Wasserflussstabilitätstests der hergestellten Beschichtungen zeigen. [32] k) und l) Veränderung des makroskopischen Aussehens der Beschichtungen in (i) bzw. (j), analysiert anhand der Pixelhelligkeit. [32] Abbildung adaptiert mit Genehmigung von Wei et al. [31] und Lee et al. [32]

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“…18,37 reported to reduce crosstalk. 36,38,39 Previous solutions to droplet crosstalk in mineral oil-based systems, such as BSA, have not been successful in mitigating crosstalk with fluorocarbon carrier fluids. 18 Investigations into droplet crosstalk have relied on spectroscopic analysis based on fluorescent, 18,[26][27][28]37,40 colorimetric, 41 or secondary reports 38,41 to determine the extent of transfer.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Direct partitioning has been largely discounted as a mechanism due to the low solubility of small molecules in fluorocarbons, but some have hypothesized that a small amount of crosstalk can still occur through semi-fluorinated oils . Recent investigations of fluorosurfactant stabilized systems have supported the view that reverse micelles are a mechanism for droplet-to-droplet transport. , These findings have led to the development of new surfactants that are reported to reduce crosstalk. ,, Previous solutions to droplet crosstalk in mineral oil-based systems, such as BSA, have not been successful in mitigating crosstalk with fluorocarbon carrier fluids …”

Section: Introductionmentioning

confidence: 99%

Evaluation of Analyte Transfer between Microfluidic Droplets by Mass Spectrometry

et al. 2023

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Droplet microfluidics enables high-throughput experimentation and screening by encapsulating chemical and biochemical samples in aqueous droplets segmented by an immiscible fluid. In such experiments, it is critical that each droplet remains chemically distinct. A common approach is to use fluorinated oils with surfactants to stabilize droplets. However, some small molecules have been observed to transport between droplets under these conditions. Attempts to study and mitigate this effect have relied on evaluating crosstalk using fluorescent molecules, which inherently limits the analyte scope and conclusions drawn about the mechanism of the effect. In this work, transport of low molecular weight compounds between droplets was investigated using electrospray ionization mass spectrometry (ESI-MS) for measurement. The use of ESI-MS significantly expands the scope of analytes that can be tested. We tested 36 structurally diverse analytes that were found to exhibit crosstalk ranging from negligible to complete transfer using HFE 7500 as the carrier fluid and 008-fluorosurfactant as a surfactant. Using this data set, we developed a predictive tool showing that high log P and log D values correlate with high crosstalk, and high polar surface area and log S correlate with low crosstalk. We then investigated several carrier fluids, surfactants, and flow conditions. It was discovered that transport is strongly dependent on all of these factors and that experimental design and surfactant tailoring can reduce carryover. We present evidence for mixed crosstalk mechanisms including both micellar and oil partitioning transfer. By understanding the driving mechanisms, surfactant and oil compositions can be designed to better reduce chemical transport for screening workflows.

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Linear triglycerol-based fluorosurfactants show high potential for droplet-microfluidics-based biochemical assays

Abstract: We decoupled the influence of different spatial geometries of oligoglycerol-based head groups and contributed to the goal of rational fluorosurfactant design for droplet microfluidics-based water-in-oil emulsions and suited applications.

Cited by 13 publications

References 30 publications

Droplet Surface Immunoassay by Relocation (D-SIRe) for High-Throughput Analysis of Cytosolic Proteins at the Single-Cell Level

Droplet Surface Immunoassay by Relocation (D-SIRe) for High-Throughput Analysis of Cytosolic Proteins at the Single-Cell Level

Grenzflächen mit fluorierten Amphiphilen: Überstrukturen und Mikrofluidik

Evaluation of Analyte Transfer between Microfluidic Droplets by Mass Spectrometry

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