Huberts Daphne H E W, Janssens Georges E, Lee Sung Sik, Vizcarra Ima Avalos, Heinemann Matthias
Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen.
J Vis Exp. 2013 Aug 20(78):e50143. doi: 10.3791/50143.
We demonstrate the use of a simple microfluidic setup, in which single budding yeast cells can be tracked throughout their entire lifespan. The microfluidic chip exploits the size difference between mother and daughter cells using an array of micropads. Upon loading, cells are trapped underneath these micropads, because the distance between the micropad and cover glass is similar to the diameter of a yeast cell (3-4 μm). After the loading procedure, culture medium is continuously flushed through the chip, which not only creates a constant and defined environment throughout the entire experiment, but also flushes out the emerging daughter cells, which are not retained underneath the pads due to their smaller size. The setup retains mother cells so efficiently that in a single experiment up to 50 individual cells can be monitored in a fully automated manner for 5 days or, if necessary, longer. In addition, the excellent optical properties of the chip allow high-resolution imaging of cells during the entire aging process.
我们展示了一种简单微流控装置的应用，在该装置中，单个出芽酵母细胞在其整个生命周期内都能被追踪。微流控芯片利用微垫阵列来利用母细胞和子细胞之间的大小差异。加载时，细胞被困在这些微垫下方，因为微垫与盖玻片之间的距离与酵母细胞的直径（3 - 4μm）相似。加载过程完成后，培养基持续冲洗通过芯片，这不仅在整个实验过程中创造了一个恒定且明确的环境，还将新出现的子细胞冲洗出去，由于子细胞尺寸较小，它们不会被困在微垫下方。该装置能非常有效地保留母细胞，以至于在单次实验中，多达50个单个细胞可以以全自动方式被监测5天，如有必要，时间还可更长。此外，芯片出色的光学特性使得在整个衰老过程中都能对细胞进行高分辨率成像。
