退半月板诱导酵母细胞在单细胞水平上在微流体通道内对接

Min Cheol Park, J. Hur, K. Kwon, Jee-Won Park, Sang-Hyun Park, K. Suh
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引用次数: 0

摘要

我们提出了一种简单的半月板诱导方法,将非粘附酵母细胞捕获到微流控通道内的微孔上。微孔是用紫外光固化聚氨酯丙烯酸酯(PUA)溶液在玻璃基板上通过毛细管模压制成的,形成了定义明确、坚固的微结构。将出芽酵母(Saccharomyces cerevisiae)的细胞悬浮液通过毛细管填充引入微流体通道,随后通过蒸发产生退缩的半月板。随着半月板的消退,一到五个酵母细胞在半月板薄区域产生的横向毛细管力自发地捕获到微孔上。利用这个细胞平台,我们通过监测绿色荧光蛋白(GFP)的表达随时间的变化,观察了酵母细胞对交配信息素(α因子)刺激的反应。观察到α -因子在刺激后60 min触发GFP表达,荧光强度持续60 min无变化
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Receding meniscus induced docking of yeast cells inside microfluidic channels at single cell level
We present a simple receding meniscus induced method to capture non-adherent yeast cells onto microwells inside microfluidic channels. Microwells were fabricated by capillary molding onto glass substrate using a UV curable polyurethane acrylate (PUA) solution, leading to well-defined, robust microstructures. A cell suspension of the budding yeast, Saccharomyces cerevisiae, was introduced into microfluidic channels by capillary filling and a receding meniscus was subsequently generated by evaporation. As the meniscus receded, one to five yeast cells were spontaneously captured onto microwells by lateral capillary force created at the thin region of the meniscus. Using this cell-based platform, we observed the response of yeast cells upon stimulation by a mating pheromone (alpha-factor) by monitoring the expression of green fluorescent protein (GFP) with time. It was observed that alpha-factor triggered the expression of GFP at 60 min after stimulation and the fluorescence intensity was sustained for additional 60 min without changes
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