METHOD AND APPARATUS FOR HIGH THROUGHPUT CELL - BASED ASSAYS FOR SCREENING AND DIAGNOSTICS

摘要

A method of performing an assay to detect and quantify changes in morphology or intracellular events in living or dead cells. The method comprises the step of presenting the cells on a surface for analysis in a single sample or array of samples and providing a means of fluorescently labelling cell structures, engulfed or associated particles or molecules contained on or within the cells. The cells are scanned with a detection system to illuminate the cells to excite fluorescence on or in the cells and obtaining a linear series of intensity values for light received therefrom at intervals of 10 microns or less across each cell to produce line amplitude data in at least one emission band. A threshold algorithm is applied to determine the beginning and end of each feature on the line amplitude data, wherein a feature is any perturbation from a determined background signal. The determined line amplitude data is processed for each feature for one or more emission bands to generate a value for at least one of area specific intensity, peak intensity, half-width, half-width specific intensity, total intensity, peak intensity, peak ratio, inflection ratio, 1D gaussian fit, 2D gaussian fit or a mathematical combination of these values. At least one generated value or values in combination is used to determine if the feature detected is or is not a cell and furthermore to characterise each cell according to its morphological state and/or the presence of an intracellular process.