Patent ID: 6716588
Filing Date: 2004-04-06
Classification: C12M,G01N,G06K,G06T

Abstract:
An automated method for analyzing cell spreading comprising:a) providing a substrate comprising an array of locations which contain multiple cells that are to be contacted with a test stimulus, wherein the cells possess at least a first luminescent reporter that is present in and can be used to identify cell cytoplasm; b) contacting the cells with the test stimulus; c) scanning multiple cells in each of the locations containing cells to acquire luminescent signals from the at least first luminescent reporter; d) creating a cytoplasmic image from the luminescent signals from the at least first luminescent reporter; e) creating a cytoplasmic mask from the cytoplasmic image; f) identifying cytoplasmic objects in the cytoplasmic mask; g) automatically measuring the following cell-based morphological features from the cytoplasmic object: i) One or more of cell area and cell perimeter; and ii) Cell shape, wherein cell shape is measured by determining one or more of cell box-fill ratio, cell height-width ratio, cytoplasm maximum width, cytoplasm minimum width, cell roundness, cell P2A which is defined as the cell perimeter, â€œPâ€ , squared divided by 4&pgr; times the cell area, and cytoplasm aspect ratio; and h) automatically determining the following well-based or field-based morphological features: i) one or more of a number of cytoplasmic objects in the field or well and an average of any of the cell-based morphological features in the field or well; and ii) a percentage of cells with any of the cell-based morphological features above a threshold value; wherein the morphological features in steps g) and h) provide a measure of cell spreading caused by the test stimulus.