Method for single-cell mass and electrophoretic mobility measurement
Name
701718635-MIT.pdf
Description
Full printable version
Size
22.39 MB
Format
Adobe PDF
Checksum (MD5)
773529c20db0897aedd5ceafb5f9d5f8
Author(s)
Dextras, Philip
Advisor(s)
Scott R. Manalis and Jongyoon Han.
Date Issued
2010
Publisher
Massachusetts Institute of Technology
Abstract
Analysis of single cells using flow cytometry techniques has created a wealth of knowledge about cellular phenomena that could not be obtained by population average measurements. As these techniques are integrated with others to increase the number of parameters that can be measured on single cells and these measurements are made more quantitative, their ability to discriminate between sub-populations of cells increases. Microfabricated sensors offer unique advantages in this area because their internal geometries can be engineered at a size scale comparable to the cell's, making them naturally well-suited for single-cell measurements. The suspended microchannel resonator (SMR) is a versatile flow cytometry platform which is capable measuring the mass of single cells with femtogram resolution. The net frequency shift of a resonant cantilever as the cell transits the fluid-filled microchannel running through it is proportional to the buoyant mass of the cell. The resonance frequency of the SMR is also highly sensitive to a cell's position along the cantilever's length. This thesis presents a new method which makes use of this property to accurately quantify the electrophoretic mobility (EPM) of cells transiting the SMR while subjected to oscillatory electric fields. Recorded resonance frequency time courses can be analyzed to extract both the buoyant mass and EPM of individual cells. This instrument has been used to simultaneously measure the EPM and buoyant masses of discrete polystyrene microspheres and Escherichia coli bacteria. As it has been applied to microspheres of known density, the integrated measurement makes it possible to compute the absolute mass and surface charge of individual microspheres. It has been shown that integrated single-microsphere mass and surface charge measurement enables differentiation of complex aqueous suspensions which is not possible by either measurement alone.
Description
Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biological Engineering, 2010.
Cataloged from PDF version of thesis.
Includes bibliographical references (p. 137-146).
Subjects
Biological Engineering.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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