Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Proc Natl Acad Sci U S A. 2010 Jan 19;107(3):999-1004. doi: 10.1073/pnas.0901851107. Epub 2009 Dec 23.
Cell growth comprises changes in both mass and volume--two processes that are distinct, yet coordinated through the cell cycle. Understanding this relationship requires a means for measuring each of the cell's three basic physical parameters: mass, volume, and the ratio of the two, density. The suspended microchannel resonator weighs single cells with a precision in mass of 0.1% for yeast. Here we use the suspended microchannel resonator with a Coulter counter to measure the mass, volume, and density of budding yeast cells through the cell cycle. We observe that cell density increases prior to bud formation at the G1/S transition, which is consistent with previous measurements using density gradient centrifugation. To investigate the origin of this density increase, we monitor relative density changes of growing yeast cells. We find that the density increase requires energy, function of the protein synthesis regulator target of rapamycin, passage through START (commitment to cell division), and an intact actin cytoskeleton. Although we focus on basic cell cycle questions in yeast, our techniques are suitable for most nonadherent cells and subcellular particles to characterize cell growth in a variety of applications.
细胞生长包括质量和体积的变化——这两个过程是不同的,但通过细胞周期协调。要理解这种关系,需要一种方法来测量细胞的三个基本物理参数:质量、体积和两者的比例,即密度。悬浮微通道谐振器可以精确测量酵母细胞的质量,精度达到 0.1%。在这里,我们使用悬浮微通道谐振器和库尔特计数器,通过细胞周期测量出芽酵母细胞的质量、体积和密度。我们观察到,在 G1/S 转换时,在芽形成之前,细胞密度增加,这与之前使用密度梯度离心的测量结果一致。为了研究这种密度增加的原因,我们监测生长中的酵母细胞的相对密度变化。我们发现,密度的增加需要能量,需要雷帕霉素靶蛋白这个蛋白质合成调节剂的功能,需要通过起始点(细胞分裂的承诺),并且需要完整的肌动蛋白细胞骨架。虽然我们专注于酵母中的基本细胞周期问题,但我们的技术适用于大多数非附着细胞和亚细胞颗粒,可用于各种应用中描述细胞生长。