Evanescent wave and video microscopy methods for directly measuring interactions between surface-immobilized biomolecules

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Title: Evanescent wave and video microscopy methods for directly measuring interactions between surface-immobilized biomolecules
Author: Everett, William Neil
Abstract: Spatial and temporal tracking of passively diffusing functionalized colloids continues to be an improving and auspicious approach to measuring weak specific and non -specific biomolecular interactions . Evidence of this is given by the recent increase in published studies involving the development and implementation of these methods . The primary aim of the work presented in this dissertation was to modify and optimize video microscopy (VM ) and total internal reflection microscopy (TIRM ) methods to permit the collection of equilibrium binding and sampling data from interaction of surface -immobilized biomolecules . Supported lipid bilayers were utilized as model systems for functionalizing colloid and wall surfaces . Preliminary results measuring calcium -specific protein -protein interactions between surface immobilized cadherin fragments demonstrate the potential utility of this experimental system and these methods . Additionally , quantum dot -modified colloids were synthesized and evanescent wave -excited luminescence from these particles was used to construct potential energy profiles . Results from this work demonstrate that colloids can be used as ultra -sensitive probes of equilibrium interactions between biomolecules , and specialized probes , such as those modified with quantum dots , could be used in a spectral multiplexing mode to simultaneously monitor multiple interactions .
URI: http : / /hdl .handle .net /1969 .1 /ETD -TAMU -1585
Date: 2009-05-15

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Evanescent wave and video microscopy methods for directly measuring interactions between surface-immobilized biomolecules. Available electronically from http : / /hdl .handle .net /1969 .1 /ETD -TAMU -1585 .

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