High precision mass-based assay to examine growth regulation of the cell cycle

Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2014.

Bibliographic Details
Main Author: Gulati, Amneet
Other Authors: Scott R. Manalis.
Format: Thesis
Language:eng
Published: Massachusetts Institute of Technology 2014
Subjects:
Online Access:http://hdl.handle.net/1721.1/90147
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author Gulati, Amneet
author2 Scott R. Manalis.
author_facet Scott R. Manalis.
Gulati, Amneet
author_sort Gulati, Amneet
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description Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2014.
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spelling mit-1721.1/901472019-04-10T13:20:24Z High precision mass-based assay to examine growth regulation of the cell cycle Gulati, Amneet Scott R. Manalis. Massachusetts Institute of Technology. Department of Biological Engineering. Massachusetts Institute of Technology. Department of Biological Engineering. Biological Engineering. Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2014. Cataloged from PDF version of thesis. Includes bibliographical references (pages 104-111). Studying biophysical properties of cells can provide insight into the metabolic mechanisms and regulation of cell cycle processes. Though size is considered to be a fundamental property of cell state, its measurement on a single-cell basis with high-resolution has been elusive primarily due to enormous experimental barriers. This thesis discusses the use of a cantilever based suspended microchannel resonator (SMR) to measure mass, and resistive pulse based Coulter counter to measure volume. First, we discuss the implementation of several engineering principles that have enabled the SMR to measure size with a high precision and temporal resolution. As a result, growth rates can now be estimated at a single-cell basis with unprecedented precision of ~170 fg.hr-¹. Second, we employ the SMR to investigate the coordination between the fundamental processes of cell growth and cell division cycle. Contrary to the reigning 60-yr old hypothesis of a deterministic size-control of the cell cycle, it is observed that cells display significant size variability at the Start checkpoint of the cell cycle. Furthermore, the measurements find only a weak size-control on the time spent in G1. Remarkably, it is observed that the cell's initial growth rate is a significantly better predictor of G1 duration than its initial size. Third, we develop a method to enable continuous, long-term volume measurement. Based on a commercial Coulter counter device, it provides a complementary technique for high-throughput measurement and continuous sampling of cell volume, as well volumetric growth rate on a population-scale. by Amneet Gulati. Ph. D. 2014-09-19T21:42:32Z 2014-09-19T21:42:32Z 2014 2014 Thesis http://hdl.handle.net/1721.1/90147 890197473 eng MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission. http://dspace.mit.edu/handle/1721.1/7582 113 pages application/pdf Massachusetts Institute of Technology
spellingShingle Biological Engineering.
Gulati, Amneet
High precision mass-based assay to examine growth regulation of the cell cycle
title High precision mass-based assay to examine growth regulation of the cell cycle
title_full High precision mass-based assay to examine growth regulation of the cell cycle
title_fullStr High precision mass-based assay to examine growth regulation of the cell cycle
title_full_unstemmed High precision mass-based assay to examine growth regulation of the cell cycle
title_short High precision mass-based assay to examine growth regulation of the cell cycle
title_sort high precision mass based assay to examine growth regulation of the cell cycle
topic Biological Engineering.
url http://hdl.handle.net/1721.1/90147
work_keys_str_mv AT gulatiamneet highprecisionmassbasedassaytoexaminegrowthregulationofthecellcycle