The long-term goal of this project is to develop a high-speed instrumentation system that differentially analyzes and sorts individual, viable, multicellular organisms (MCOs) such as nematodes, fruit fly larvae, and zebrafish embryos. Capabilities of this system include the preparation and distribution of homogeneous populations of MCOs from mixed populations and the identification and sorting of phenotypic variants. Potential applications include therapeutic/toxic compound screening and functional genomics. Currently, there are no automated systems for this purpose, consequently progress in the laboratory is labor intensive, slow and non-quantitative. The proposed high-speed system will overcome these barriers to progress. A novel flow-sorting system has been constructed to demonstrate the feasibility of this technology using nematodes and length as a single, phenotypic parameter. The system automatically selects nematodes by length (indicative of development stage or mutant type) from mixed populations and distributes them unharmed in microplates at a rate of 20 nematodes per second. These successful studies have revealed the need for more detailed morphological parameters to classify and select mutants.
Our specific aims for Phase I are to construct and characterize a system that would monitor the nematode along its axis and report quantitative morphological features correlated to axial position.
This research will result in commercial instrumentation systems to facilitate the use of multicellular organisms in drug discovery research and high throughput screening. These systems will speed the process of identifying and sorting disease-related mutants from larger populations. They will be capable of monitoring large numbers of MCOs in short times, therefore providing high quality statistical interpretation of the effect of therapeutic compounds on gene expression.
