Instrumental advancements to improve sample throughput for aminopeptidase profiling
Abstract
Microorganism identification is one area of science where rapid and accurate identification is imperative. Aminopeptidase profiling is a method that uses metabolic characteristics of microorganisms as a means for identification. Unfortunately, long incubation times, difficult experimental procedures, and extensive data collection periods have made this technique impractical for routine microorganism identification. In order to improve the efficiency of aminopeptidase profiling, a fluorimeter that combines the selectivity of time-resolved detection and the sample throughput of a 96-well plate reader has been developed. The time-resolved 96-well plate reader allows the sample loading, data collection and analysis for a twenty amino acid profile of four samples to be completed in 30 min. Compared to the 3.5 hrs required by comparable aminopeptidase techniques, this instrument represents a significant improvement in sample throughput. Research has continued in an attempt to develop libraries of aminopeptidase profiles to be used as standards to identify unknown microorganisms. The future of aminopeptidase profiling will be enhanced further by the development of a multiple channel, microfabricated capillary electrophoresis device. By moving to the microfabricated device, we will drastically reduce the sample volumes which, in turn, will reduce total cell concentrations and incubation times. Unfortunately, it is presently difficult to detect multiple channels in parallel on the microfabricated device due to the macroscopic nature of the spectroscopic detection system. One possible solution is to fabricate channels that will guide the light at the same time the separation system is created. This would bring the detection system to the microscopic scale of the separation system. Once proven successful, it will be possible to create a device with multiple separation channels and waveguides to be used for the detection system.
Degree
Ph.D.
Advisors
Lytle, Purdue University.
Subject Area
Analytical chemistry|Microbiology|Public health
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