Smart Microscope System for Radiation Biodosimetry

An automated, digitally-controlled microscopy system for improving accuracy of estimation of radiation exposure by biodosimetry in a sample consisting of cells prepared for cytogenetic analysis from a single individual, said method comprising:

• (i) using a microscope equipped with a computer controlled digital camera to sequentially acquire images of cells containing metaphase chromosomes,
• (ii) performing a digital analysis of objects in each image to either select or reject the image for radiation exposure determination, based on one or more properties of segmented objects contained therein, said properties including object count, length, width, contour finite difference, centromere density,
• (iii) directing the microscope system to discontinue collecting images on a sample of metaphase cells after a sufficient number of images have been captured to determine radiation dose,
• (iv) classifying likely dicentric chromosomes in the set of selected digital images of cells in the metaphase stage of the cell cycle,
• (v) determining which of the likely dicentric chromosomes in said digital image are not true dicentric chromosomes using segmentation procedures that discriminate true positive dicentric chromosomes from other objects,
• (vi) eliminating false positive dicentric chromosomes from the set of likely dicentric chromosomes the same digital images,
• (vii) correcting the count of the dicentric chromosomes in each digital image by subtracting the number of false positive dicentric chromosomes from the total likely dicentric chromosomes in an image,
• (viii) determining the dose response for the sample, which is average dicentric chromosome frequency over all images from that sample, by summing the total number of corrected dicentric chromosomes in all images from the same sample and dividing by the number of images,
• (ix) computing the radiation exposure, Y, using a previously determined calibration curve that is related to the dose response, X, by the quadratic equation, Y=aX2+bX+c, where a, b, and c are the coefficients of the curve,
• (x) sending a signal to the digitally controlled microscope system indicating that the process of collecting images from a sample has been completed, and terminating the collection of new image data for that sample.