Rosebrock Adam P
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
Cold Spring Harb Protoc. 2017 Jan 3;2017(1):2017/1/pdb.prot088732. doi: 10.1101/pdb.prot088732.
In yeast, cell size is normally tightly linked to cell cycle progression. Centrifugal elutriation is a method that fractionates cells based on the physical properties of cell size-fluid drag and buoyant density. Using a specially modified centrifuge and rotor system, cells can be physically separated into one or more cohorts of similar size and therefore cell cycle position. Small G daughters are collected first, followed by successively larger cells. Elutriated populations can be analyzed immediately or can be returned to medium and permitted to synchronously progress through the cell cycle. This protocol describes two different elutriation methods. In the first, one or more fractions of synchronized cells are obtained from an asynchronous starting population, reincubated, and followed prospectively across a time series. In the second, an asynchronous starting population is separated into multiple fractions of similarly sized cells, and each cohort of similarly sized cells can be analyzed separately without further growth.
在酵母中，细胞大小通常与细胞周期进程紧密相关。离心淘析是一种根据细胞大小的物理特性（流体阻力和浮力密度）对细胞进行分级分离的方法。使用经过特殊改装的离心机和转子系统，细胞可以根据大小被物理分离成一个或多个大小相似的群体，从而处于相似的细胞周期阶段。首先收集较小的G1期子细胞，随后依次收集更大的细胞。淘析得到的细胞群体可以立即进行分析，也可以放回培养基中，使其同步进行细胞周期进程。本方案描述了两种不同的淘析方法。第一种方法是从异步起始群体中获得一个或多个同步化细胞组分，重新孵育，并在一个时间序列中进行前瞻性跟踪。第二种方法是将异步起始群体分离成多个大小相似的细胞组分，每个大小相似的细胞群体无需进一步生长即可单独进行分析。
