Segmentation Of Nuclei In Confocal Image Stacks Using Performance Based Thresholding

Combinatorial Transcriptional Fluorescent In Situ Hybridization (CT-FISH) is a confocal fluorescence imaging technique enabling the detection of multiple active transcription units in individual interphase diploid nuclei. As improved combinatorial labeling methods allow simultaneous measurement of gene activities to expand from five genes in a single embryo or tissue section to upward of twenty genes, transforming image stacks into usable data becomes an increasingly labor intensive task. In this paper we describe our progress towards a method for the computational analysis of confocal images from Drosophila melanogastar that involves the segmentation of the cell nuclei and of nascent transcription sites of specific genes. Using image processing and machine learning algorithms, we allow experimentalists to reiteratively tune and improve the analysis system to reflect biological reality.