Reliable yeast replicative lifespan determination with a single-channel microfluidic chip.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-10-31 DOI:10.1242/bio.060596

Valentina Salzman, Moises R Bustamante Torres, Francisco G Correa Tedesco, Nahuel Tarkovski, María J Godás Willems, Joaquín N Bravo, Magalí Mercuri, Dante G Mercado, Guido Berlin, Martín G Bellino, Pablo S Aguilar, Laura C Estrada

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Abstract

Saccharomyces cerevisiae is a powerful model for aging research due to its short lifespan and genetic malleability. Microfluidic devices offer an attractive approach enabling rapid monitoring of hundreds of cells during their entire replicative lifespan (RLS). Yet, key operational issues such as contaminations, cell loss, and cell aggregates-dependent flow obstruction can hinder RLS experiments. We report the development of a microfluidic device configuration that effectively prevents flow blockage. We conducted comprehensive performance characterization, evaluating trapping efficiency, cell retention, budding orientation, and cell aggregate formation. The optimized device successfully supported long-term culturing and reliable RLS measurements of budding yeast strains. For accurate lifespan determination, a detailed workflow is provided that includes device fabrication, live microscopy setup, and characterization of cell age distribution. This work describes an accessible and reliable microfluidic device for yeast RLS studies, promoting further exploration in aging research.

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利用单通道微流控芯片可靠测定酵母的复制寿命

毕赤酵母（Saccharomyces cerevisiae）寿命短，基因可塑性强，是研究衰老的有力模型。微流控装置提供了一种极具吸引力的方法，可在数百个细胞的整个复制寿命（RLS）期间对其进行快速监测。然而，一些关键的操作问题，如污染、细胞丢失和细胞聚集导致的流动阻塞，都会阻碍 RLS 实验的进行。我们报告了一种能有效防止流动阻塞的微流体设备配置的开发情况。我们进行了全面的性能表征，评估了捕获效率、细胞保留率、出芽定向和细胞聚集的形成。优化后的装置成功支持了长期培养和可靠的出芽酵母菌株 RLS 测量。为了准确测定细胞寿命，本文提供了一个详细的工作流程，包括设备制造、活体显微镜设置和细胞年龄分布表征。这项工作描述了一种用于酵母 RLS 研究的简便可靠的微流控装置，促进了衰老研究的进一步探索。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.

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