Photomicrographic image-processing method for automatic scanning, detection and classification of particles

A photomicrographic image-processing method for automatic scanning, detection and classification of particles present in a sample presented for photomicrography, comprising—

• a) Constituting an application environment by communicatively associating an optical microscope having a motorized stage to a computer, wherein—
• The photomicrographic image-processing method for automatic scanning, detection and classification of particles present in a sample presented for photomicrography is provisioned for execution, as an executable software, on said computer;
• and
• the optical microscope is outfitted with a digital camera for capturing images from the field of view of said microscope and relaying said captured images in real time to said computer for processing by the executable software provisioned on said computer.
• b) Defining at instance of the user via a computer user interface of the executable software, a set of scanning parameters being opted among particle size, analysis area, scan position, captured image resolution, and magnification;
• c) Marking at instance of the user, via a computer user interface of the executable software simulating an image of the slide, a rectangular area to be scanned;
• d) In accordance with logic of the executable software, causing the execution of an AutoScanner subroutine programmed within the executable software, said subroutine comprising the steps of—
• Computing the actual number of fields in terms of rows and columns, to be captured in the rectangular area marked for scanning according to the magnification and captured image resolution;
• Marking the actual initial position to therefore move the scanner X and Y position on the slide;
• Causing the microscope stage to move in Z direction in steps to therein capture multiple images at different Z positions;
• Computing the optimum sharpness of the images and sets the Z movement at the most focused position;
• Saving the captured image in memory of the computer in a shared folder with its corresponding row and column position in the image name.
• e) In accordance with the defined set of scanning parameters, causing at least one image to be captured from the microscope corresponding to the analysis area and magnification selected, therein saving the at least one image to a ready shared folder in memory of the computer with its filename corresponding to the scan position selected;
• f) In accordance with logic of the executable software, causing the execution of a validation subroutine, said subroutine comprising the steps of—
• Determining first, whether or not the current position is the Last X position to thereby move the motorized slide stage in Y direction in the event the determination is positive, and alternatively reverting to the basal program if said determination is negative.
• Determining first, whether or not the current position is the Last Y position to thereby read the image from shared folder of the computer in the event the determination is positive, and alternatively reverting to the basal program if said determination is negative.
• g) Upon the sequential determinations in step f) being positive, preprocessing the image from the shared folder by smoothening said at least one image for removal of noise.
• h) Identifying contours in said at least one image, therein selecting contours of same gray value variation to form contour groups and determining, among said contour groups, the best contours on basis of the predefined parameters for object identification, in a downstream process comprising—
• Once objects are identified, computing feature data corresponding to said objects on basis of their size, shape, color, and texture;
• Applying at least one filter on basis of size, sharpness, and agglomeration to the feature data generated to remove artifacts and result in filtered object data;
• Determining whether the filtered object data corresponds to objects present on the left and top boundaries of the image under processing, and based on this determination, causing the execution of a suitable sub-process for resulting in either between boundary particle identification and particle classification respectively;
• Determining whether the image being processed is the last image of total fields, and based on this determination, causing the execution of a suitable sub-process for either between termination with computation of result statistics and termination of the execution of the executable software upon reaching threshold of the user-defined parameter values respectively.
• i) In accordance with logic of the executable software, causing the execution of a validation subroutine to determine whether or not the image being processed is last image of total fields to thereby allow the executable software to compute and present the result statistics of the analysis undertaken in the event said determination is positive, and alternatively repeating steps g) to h) until a discernable accurate resultant image is obtained.