BRIEF RESEARCH REPORT article
Front. Bioeng. Biotechnol.
Sec. Nanobiotechnology
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1568027
This article is part of the Research TopicMicrodroplets and the Art of Directed Evolution: Pitfalls and Promising AdvancesView all articles
A sticky situation -Simple method for rapid Poissonian encapsulation of highly aggregation-prone microbeads in polydisperse emulsions
Provisionally accepted
- Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
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Directed evolution leverages the principles of natural selection to engineer biomolecules with desired properties. Microbead-based approaches within water-in-oil emulsions have proven invaluable for high-throughput in vitro selections. However, highly aggregation-prone microbeads present significant challenges, including clustering, inconsistent distribution, and droplet instability.Here, we introduce a simple and cost-effective method for generating polydisperse emulsions with restored Poissonian distributions of highly aggregation-prone microbeads. This approach utilizes modified gel loader pipette tips, drawn out to create nozzles capable of disrupting microbead clusters during emulsification. Two widely utilized oil-surfactant formulations -mineral oil with Abil EM 90 and FluoSurf in HFE 7500 -were evaluated for emulsion preparation. Emulsions prepared using the modified nozzles exhibited exceptional stability, maintaining integrity during week-long incubations at 37°C, and reliably distributed microbeads into droplets in accordance with a Poissonian distribution despite the microbeads' highly aggregation-prone property.
Keywords: Microbeads, In vitro compartmentalization, Emulsion, encapsulation, Aggregation, clustering, Poisson Distribution
Received: 28 Jan 2025; Accepted: 16 Jun 2025.
Copyright: © 2025 Hasecke and Hoyer. 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) or licensor 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.
* Correspondence: Filip Hasecke, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
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