Methods and systems for identifying well wall boundaries of microplates

An apparatus for locating wall boundaries of test wells arranged on the surface of a microplate comprising:

• an imaging microscope having at least one objective lens to acquire images at one or more magnifications and a stage to hold the microplate;
• an excitation source to illuminate the microplate on the stage;
• a digital image device connected to the microscope to acquire a digital image of the microplate;
• a storage device in communication with the digital image device capable of storing the digitized images of the microplate;
• and
• a processor in communication with the storage device and capable of;
• analyzing the digital image of the microplate utilizing, at least in part, pixel intensity information to detect features of the wall boundaries of the test wells;
• generating a candidate edge image of the microplate using the features of the wall boundaries wherein the candidate edge image represents locations of one or more segments of the wall boundaries;
• analyzing the candidate edge image using an algorithm to calculate the outer perimeter boundary of one or more test wells and to determine the spatial location of the outer perimeter boundary on the surface of the microplate;
• analyzing the outer perimeter boundary of the one or more test wells utilizing, at least in part, pixel intensity information to determine the spatial location of the inner perimeter boundary of one or more test wells;
• generating a binary image of the microplate wherein the binary image is a spatial domain image of the location of the inner and outer boundaries of the test wells;
• and
• mapping the spatial domain image on the microplate to identify test well regions on the surface of the microplate.