Fluorescent dot counting in interphase cell nuclei

Abstract

Fluorescence in situ hybridization allows the enumeration of chromosomal abnormalities in interphase cell nuclei. This process is called dot counting. To estimate the distribution of chromosomes per cell, a large number of cells have to be analysed, particularly when the frequency of aberrant cells is low. Automation of dot counting is desirable because manual counting is tedious, fatiguing, and time consuming. We have developed a completely automated fluorescence microscope system that counts fluorescent hybridization dots for one probe in interphase cell nuclei. This system works with two fluorescent dyes-one for the DNA hybridization dots and one for the cell nucleus. A fully automated scanning procedure has been used for the image acquisition. After an image is acquired it has to be analysed in order to find the nuclei and to detect the dots. This article focuses upon the dot detection procedure. Three different algorithms are presented. The problems of 'overlapping' dots and split dots are discussed. The automated dot counter has been tested on a number of normal specimens where DAPI was used for the nucleus counter stain and a centromeric probe was used to mark the chromosome 12. The slides contained lymphocytes from cultured blood. The performance of the different algorithms has been evaluated and compared with manually obtained results. The automated counting results approximate the results of manual counting.

Intelligent Screening Systems for Cervical Cancer

“…Several researchers have identified capability of the size and shape features to classify the cervix using the cytology image [ 72 , 73 , 75 ], FISH image [ 60 , 76 , 77 ], and electromagnetic spectrum [ 24 , 78 ]. Besides these features, the ratio of the same feature for different parts of a biological structure is used as another feature.…”

“…For FISH image, the features from labeled biomarker spots of chromosomes 3 (red spot) and X (green spot) are the size of each colored spot [ 60 , 76 , 77 ], the effective radius of each red or green spot computed as the radius of a circle that had the same size as the colored spot [ 60 , 76 , 77 ], and the circularity of each colored spot [ 60 , 77 ].…”

“…The features are applied for cellular-level case. The specific features implemented for cytology images are distribution of cell [ 72 ] and distribution of the nucleus [ 72 ], while the distances between the same color spots [ 60 , 77 ], the distance between the centers of the two spots [ 60 , 77 ], the gravity center of each colored spot [ 60 ], and the total number of red spots and green spots [ 60 , 76 , 77 ] have been implemented in the FISH images. The thresholding techniques are applied to extract the features in the cellular-level case [ 60 , 72 ].…”

Automated detection and analysis of fluorescent in situ hybridization spots depicted in digital microscopic images of Pap-smear specimens

“…These studies indicated a strong correlation of detection results between the manual methods and the semiautomated computer schemes. 19,20 Because the semiautomated methods require userintervention to exclude poorly segmented, overlapped, clustered, or nonepithelial cells and therefore are impractical in the clinical practice, some fully automated schemes have recently been developed and tested. These schemes usually include two processing and detection steps.…”