Analytical Measurements of Single Cells and Cellular Organelles

Fuller, Robert Ray

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Description

Title

Analytical Measurements of Single Cells and Cellular Organelles

Author(s)

Fuller, Robert Ray

Issue Date

1998

Doctoral Committee Chair(s)

Sweedler, Jonathan V.

Department of Study

Chemistry

Discipline

Chemistry

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

Biology, Neuroscience

Language

eng

Abstract

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.

Graduation Semester

1998

Type of Resource

text

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