Techniques for Analysis, Sorting, and Dispensing of C. elegans on the COPAS™ Flow-Sorting System

Abstract

The COPAS™ Biosorter is a flow cytometer designed to accommodate large objects the size of Caenorhabditis elegans. This instrumentation brings high-speed automated analysis and sorting to this small model organism. The Biosort system optically analyzes and sorts living multicellular organisms on the basis of fluorescent protein expression patterns and other optical signatures, at rates up to about 100 organisms per second. The Biosort is capable of fluorescently analyzing and sorting multicellular organisms that are many-fold larger than single cells. Animals pass through a laser beam focused to the center of the flow cell. This beam is narrower than the animal so that multiple measurements are made per animal, which means that the organism is optically scanned along its long axis as it flows. Stable laminar flow in the flow cell acts to orientate the animal with the flow stream. Fluorescent locations along the axis of the animal are sequentially excited as the organism flows through the line of focus. The fluorescent properties of commonly used reagents in the research field allow the user to detect fluorescent protein expression, lectin and antibody binding, and autofluorescence. The ability to dispense organisms as they emerge from the flow cell allows for the collection of those organisms that have certain optical properties defined by the researcher. Also, dispensing allows for the precise distribution of specific numbers of animals for analysis that can vary with organism numbers.