Analytical Measurements of Single Cells and Cellular Organelles

While our laboratory and others have the ability to assay the contents of single neurons, far more interesting is the assay single vesicles within those neurons. Since a possible means of neuronal information encoding may be due to the localization of different vesicle populations in different areas of a single cell, this work may have fundamental importance. To this end, background information and experiments involving the isolation of neuropeptide-containing biological vesicles are presented. Evidence of vesicle fusion-enhanced derivatization is shown to be an effective method of increasing the overall yield of derivatized molecules contained within synthetic vesicles. While this technique was not transferable to biological vesicle systems, a flow cytometry method of single vesicle analysis is demonstrated. The flow cytometry technique is shown to be quantitative for the sizing of submicron, fluorescently-labeled synthetic vesicles. It is hoped that extension of the flow cytometry method to the native fluorescence detection of individual biological vesicles will yield spectral patterns characteristic of vesicle contents.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Chemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Fuller, Robert Ray

Contributors dc:contributor

• Sweedler, Jonathan V.

• Biology, Neuroscience

Language dc:language

eng

Identifier

(MiAaPQ)AAI9904459

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/84410