single cell analysis of yeast replicative aging using a new generation of microfluidic device单细胞分析酵母复制衰老使用新一代的微流体装置.pdfVIP

Single Cell Analysis of Yeast Replicative Aging Using a New Generation of Microfluidic Device 1,2 1 1,2 1 3 1,4 2 Yi Zhang , Chunxiong Luo , Ke Zou , Zhengwei Xie , Onn Brandman , Qi Ouyang *, Hao Li * 1The State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, Center for Quantitative Biology, and School of Physics, Peking University, China, 2 Department of Biochemistry and Biophysics and California Institute for Quantitative Biosciences, University of California San Francisco, San Francisco, California, United States of America, 3 Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California, United States of America, 4 Department of Physics, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China Abstract A major limitation to yeast aging study has been the inability to track mother cells and observe molecular markers during the aging process. The traditional lifespan assay relies on manual micro-manipulation to remove daughter cells from the mother, which is laborious, time consuming, and does not allow long term tracking with high resolution microscopy. Recently, we have developed a microfluidic system capable of retaining mother cells in the microfluidic chambers while removing daughter cells automatically, making it possible to observe fluorescent reporters in single cells throughout their lifespan. Here we report the development of a new generation of microfluidic device that overcomes several limitations of the previous system, making it easier to fabricate and operate, and allowing functions not possible with the previous desig