Letourneau Alexander, Kegel Jack, Al-Ramahi Jehad, Yachinich Emily, Krause Harris B, Stewart Cameron J, McClean Megan N
Department of Biomedical Engineering, University of Wisconsin-Madison, United States.
MethodsX. 2020 Apr 24;7:100891. doi: 10.1016/j.mex.2020.100891. eCollection 2020.
Traditional methods to assess microbial cells during suspension culture require laborious and frequent manual sampling. Approaches to automate sampling and assessment utilize dedicated, sophisticated equipment and suffer from a lack of temporal resolution and sampling efficiency. In this study we describe a simple microfluidic device that allows microbial cells to be sampled from suspension culture and rapidly slowed and concentrated for single-cell imaging on a standard laboratory microscope. We demonstrate a device that: •slows and concentrates microbial cells, specifically budding yeast, sampled from suspension culture and improves imaging of individual cells by concentrating them in a single focal plane•provides imaging quality and temporal resolution that is capable of monitoring dynamic spatiotemporal processes, such as nuclear localization of a protein•is inexpensive and simple enough to be fabricated and used in laboratories equipped for standard molecular and cellular biology.
在悬浮培养过程中评估微生物细胞的传统方法需要费力且频繁地手动采样。自动化采样和评估方法使用专门的复杂设备,且存在时间分辨率和采样效率不足的问题。在本研究中,我们描述了一种简单的微流控装置,该装置能够从悬浮培养物中对微生物细胞进行采样,并快速使其减速和浓缩,以便在标准实验室显微镜下进行单细胞成像。我们展示了一种装置,该装置:• 使从悬浮培养物中采样的微生物细胞(特别是出芽酵母)减速并浓缩,并通过将它们集中在单个焦平面中来改善单个细胞的成像 • 提供能够监测动态时空过程(如蛋白质的核定位)的成像质量和时间分辨率 • 价格低廉且足够简单,可在配备标准分子和细胞生物学设备的实验室中制造和使用。