Measurement of mass, density, and volume during the cell cycle of yeast.
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.
Bryan
Andrea K
AK
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Goranov
Alexi
A
Amon
Angelika
A
Manalis
Scott R
SR
D000199:Actins / Q000378:metabolism; D002453:Cell Cycle; D005434:Flow Cytometry; D015003:Yeasts / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism
D000199:Actins