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Amphipathic Fluorophores for Probing Cell Membrane
Properties
Demetrius CummingsUniversity of California, Riverside
Background…
• Prestin
• Enhances electromotility
• Sound reception
• Other properties?
http://med.stanford.edu/ohns/research/
Fluorophores…
• Fluorescent component; multiple types
• Often used in diagnostics: tissues, macromolecules, diseases
http://exobio.ucsd.edu/MOD/detection.htm
The Goals…
• Synthesize an amphipathic dye for the purpose of binding to the plasma membrane.
• Monitor the photophysical properties of the dye and HEK cells in various conditions.
The Process…(part 1)
Dye preparation Utilize HEK cells
Analysis: photophysicalproperties
Combine dye and cells
Analysis: photophysicalproperties
Analysis: photophysicalproperties
Dye preparation Utilize HEK cells
The Process…(part 2)
Dye preparation Utilize HEK cells
Analysis: photophysicalproperties
Combine dye and cells
Analysis: photophysicalproperties
Analysis: photophysicalproperties
The Process…(part 3)
Dye preparation Utilize HEK cells
Analysis: photophysicalproperties
Combine dye and cells
Analysis: photophysicalproperties
Analysis: photophysicalproperties
The Process…(part 4)
Dye preparation Utilize HEK cells
Analysis: photophysicalproperties
Combine dye and cells
Analysis: photophysicalproperties
Analysis: photophysicalproperties
Summary…• The efficiency and mechanism of the
dye are halide-dependent.
• The absorbance varies directly with dye-solvent and cell-solvent concentration; the specific wavelengths of absorbance remains constant.
• Blue shift of the dye-HEK cell mix indicates binding to the less polar environment.
Future Research…
• Continue fluorescence analysis of HEK cells: excitation, emission, anisotropy
• Effect of dye on cells?
• Apply same procedure to cells transfected with prestin