FAST-FISH DETECTION AND SEMIAUTOMATED IMAGE-ANALYSIS OF NUMERICAL CHROMOSOME-ABERRATIONS IN HEMATOLOGICAL MALIGNANCIES

A new fluorescence in situ hybridization (FISH) technique called Fast- FISH in combination with semi-automated image analysis was applied to detect numerical aberrations of chromosomes 8 and 12 in interphase nuc lei of peripheral blood lymphocytes and bone marrow cells from patient s with acute myelogenous leukemia (AML) and chronic lymphocytic leukem ia (CLL). Commercially available alpha-satellite DNA probes specific f or the centromere regions of chromosome 8 and chromosome 12, respectiv ely, were used. After application of the Fast-FISH protocol, the micro scopic images of the fluorescence-labelled cell nuclei were recorded b y the true color CCD camera Kappa CF 15 MC and evaluated quantitativel y by computer analysis on a PC. These results were compared to results obtained from the same type of specimens using the same analysis syst em but with a standard FISH protocol. In addition, automated spot coun ting after both FISH techniques was compared to visual spot counting a fter standard FISH. A total number of about 3,000 cell nuclei was eval uated. For quantitative brightness parameters, a good correlation betw een standard FISH labelling and Fast-FISH was found. Automated spot co unting after Fast-FISH coincided within a few percent to automated and visual spot counting after standard FISH. The examples shown indicate the reliability and reproducibility of Fast-FISH and its potential fo r automatized interphase cell diagnostics of numerical chromosome aber rations. Since the Fast-FISH technique requires a hybridization time a s low as 1/20 of established standard FISH techniques, omitting most o f the time consuming working steps in the protocol, it may contribute considerably to clinical diagnostics. This may especially be interesti ng in cases where an accurate result is required within a few hours.