Determining an optimal intensity threshold for use in image analysis of a stained biological specimen

摘要

The method comprises: a- Obtaining digital image(s) of a stained biological specimen 100; b- Using a first illumination intensity 110 to determining a skeleton mask for the digital image 120; c- Calculating a fitness value for the skeleton mask 130, where the fitness value depends on the number of pixels represented by the mask; d- Sorting the first illumination intensity (threshold) value and fitness value 140; e- Repeating the steps b-d for plurality of further predetermined illumination intensity (threshold) values and f- Determine the optimal intensity (threshold) value 160 that has the highest fitness value stored in step d. The mask may be one pixel in thickness, and the skeletonising of the identified areas may involve defining a median line through the identified areas. The staining may be associated with a boundary of a cell, and the fitness value may dependent on one or more of: number of pixels in the skeleton mask, number of closed loops in the mask, or the size or shapes of such loops. A method for delineating areas on biological specimen images may be based on the method.