Method of assaying shape and structural features in cells

The method for assaying shape features of cells and determining whether the shape features are altered by exposure of the cells to a pharmaceutical or biological agent, wherein one or more of the steps are executed by an instruction set encoded as a program in software in a high-speed computer, comprising the steps of:

• (a) culturing at least two samples of a cell population, a first treated sample population being cultured in the presence of a test agent and a second untreated sample population being cultivated in the absence of the test agent,
• (b) generating a high-resolution image of each cell to be sampled from each of the sample populations as representative of the cultures, which image presents high contrast information in at least one contour of the cell or a portion of the cell, wherein the high contrast is generated at the cell edge or at least one or more interference contours, and the contrast is augmented by imaging processing,
• (c) extracting at least one contour from each cell or portion of a cell sampled from each of the sample populations,
• (d) extracting at least one boundary of at least one contour from each cell or portion of a cell that was sampled from each of the sample populations,
• (e) determining area, perimeter, and obtaining an equation for ellipse of concentration of the cell or portion of the cell and determining at least one or more variables representing shape features of the boundary or boundaries extracted from the contour or contours of one cell or portion of one cell,
• (f) repeating steps a–e for each cell sampled from each of the sample populations, and
• (g) determining whether any two populations of cells sampled, with and without exposure to test agent, differed from one another;
• wherein one or more of the steps are executed by an instruction set encoded as a program in software in a high-speed computer, comprising:
• (h) focusing of the cell or a portion of the cell at the level of at least one contour while an image is captured in a charge-coupled device or video camera and written in a recognized software format in computer memory,
• (i) processing a single image or a series of images of the cell or portion of the cell by at least one of a series of image processing algorithms including edge enhancement and threshoiding,
• (j) separating pixels representing a contour or repeated contours of the cell or portion of the cell from the pixels representing background,
• (k) extracting a boundary on the contour or contours from among the pixels representing an image of the contour(s) of a cell or portion of a cell,
• (l) determining values of an equation of ellipse of concentration and at least one dimensioned variable and determining the value of at least one dimensionless shape variable reflecting shape features of at least one boundary on the cell or portion of the cell,
• (m) combining the values of the variables into a data set representative of at least a sampled population from the treated and untreated groups, and
• (n) determining whether the cells of the unknown and known samples show a significant difference.