Open AccessAutomated FISH spot counting in interphase nuclei: Statistical validation and data correction
Author(s) -
de Solórzano Carlos Ortiz,
Santos Andrés,
Vallcorba Isabel,
GarcíaSagredo JoséMiguel,
del Pozo Francisco
Publication year - 1998
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19980201)31:2<93::aid-cyto4>3.0.co;2-j
Subject(s) - interphase , fluorescence in situ hybridization , fish <actinopterygii> , chromosome , calibration , biology , biological system , computer science , mathematics , genetics , statistics , fishery , gene
The evaluation of an automated system for Fluorescence In Situ Hybridization (FISH) spot counting in interphase nuclei is presented in this paper. Different types of experiments have been performed with samples from known populations. In all of them the goal is to detect mosaicism of chromosome X in leukocytes from mixtures in known proportions of healthy male and female blood. First the initial results from the automatic FISH analysis system were obtained and evaluated. Then the analysis was modified to reduce systematic errors, so that the results are closer to what an experienced human operator would have obtained (system calibration step). Finally, an additional control probe of chromosome Y was used to detect and discard cells where incorrect hybridization or other abnormal situations had occurred. In each step the system sensitivity was determined by the use of two statistical validation tests, so that the improvement brought about by the correction methods could be assessed. The results obtained in the study showed that, using both corrections, the system is able to detect 10% monosomies with a significance level α = 0.1%. Cytometry 31:93–99, 1998. © 1998 Wiley‐Liss, Inc.