Rapid analysis of gene copy numbers by FISH and automated spot counting from stacked confocal images Conference Paper uri icon

abstract

  • Fluorescent in situ hybridization (FISH) is an excellent method for detection of gene copy number alterations in cancer and other diseases. A limitation of the technology is the tedious, inaccurate and often highly subjective spot counting. For a number of reasons, automation of FISH spot counting has not been accomplished. FISH signals are often at different focal planes, resulting in interfering out-of-focus light. This paper describes current progress towards automated FISH spot counting, with particular reference to the previous technical limitations. A confocal digital imaging system was used to acquire a series of up to twenty-four consecutive confocal fluorescent images (along the Z-axis) of the 4m thick tumor tissue sections. Dual-color FISH with a test probe and a reference probe labeled in two different colors was applied. A new algorithm was applied that identified the spots on each level along the Z-axis, and then differentiated spots located vertically above the X-Y plane. The ratio of test to-reference probe signals produced an estimate of the gene copy number alterations in the specimens.

name of conference

  • Molecular Imaging: Reporters, Dyes, Markers, and Instrumentation

published proceedings

  • MOLECULAR IMAGING: REPORTERS, DYES, MARKERS, AND INSTRUMENTATION

author list (cited authors)

  • Grigoryan, A. M., Dougherty, E. R., Bubendorf, L., Kononen, J., & Kallioniemi, O.

citation count

  • 0

complete list of authors

  • Grigoryan, AM||Dougherty, ER||Bubendorf, L||Kononen, J||Kallioniemi, O

editor list (cited editors)

  • Bornhop, D. J., & Licha, K.

publication date

  • April 2000