System for cell-based screening

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, “P”, squared divided by 4π 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.