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Fluorescence correlation spectroscopy: ultrasensitive detection in clear and turbid media

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Title: Fluorescence correlation spectroscopy: ultrasensitive detection in clear and turbid media
Author(s): Tahari, Abdel Kader
Department / Program: Physics
Discipline: Physics
Degree: Ph.D.
Genre: Dissertation
Subject(s): fluorescence ultrasensitive turbid media somatic protein aggregates multiple-species discrimination oligomeric molecular biomaker cerebrospinal data filtration FCS PCH Method cytometry amyloid oligomer amyloid
Abstract: In this work, I describe the development of a simple, inexpensive, and powerful alternative technique to detect and analyze, without enrichment, extremely low concentrations of cells, bacteria, viruses, and protein aggregates in turbid fluids for clinical and biotechnological applications. The anticipated applications of this technique are many. They range from the determination of the somatic cell count in milk for the dairy industry, to the enumeration and characterization of microorganisms in environmental microbiology and the food industry, and to the fast and ultrasensitive detection of protein aggregates for the diagnosis of Alzheimer’s and other neurodegenerative diseases in clinical medicine. A prototype instrument has been built and allowed the detection and quantification of particles down to a few per milliliter in short scanning times. It consists of a small microscope that has a horizontal geometry and a mechanical instrument that holds a cylindrical cuvette (1 cm in diameter) with two motors that provide a rotational and a slower vertical inversion motions. The illumination focus is centered about 200 μm from the wall of the cuvette inside the sample. The total volume that is explored is large (~ 1ml/min for bright particles). The data is analyzed with a correlation filter program based on particle passage pattern recognition. I will also describe further work on improving the sensitivity of the technique, expanding it for multiple-species discrimination and enumeration, and testing the prototype device in actual clinical and biotechnological applications. The main clinical application of this project seeks to establish conditions and use this new technique to quantify and size-analyze oligomeric complexes of the Alzheimer’s disease β-peptide in cerebrospinal fluid and other body fluids as a molecular biomarker for persons at risk of Alzheimer’s disease dementia. The technology could potentially be extended to the diagnosis and therapeutic monitoring of other protein neurologic diseases where oligomeric forms may play a role.
Issue Date: 2006
Genre: Dissertation / Thesis
Type: Text
Language: English
URI: http://hdl.handle.net/2142/35266
Rights Information: 2006 Tahari ©
Date Available in IDEALS: 2012-11-27
 

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