Thick-section fluorescence in situ hybridization on formalin-fixed, paraffin-embedded archival tissue provides a histogenetic profile.
Fluorescence in situ hybridization has become a major tool for analysis of gene and chromosome copy number in normal and malignant tissue. The technique has been applied widely to fresh tissue and dispersed formalin-fixed, paraffin-embedded archival tissue, but its use on sections of archival tissue has largely been limited to sections < 6 mu thick. This does not provide intact, uncut nuclei for accurate analysis of gene or chromosome copy number. We report here a method of hybridization to sections > 20 microns thick that overcomes these difficulties. Key developments were the use of DNA probes directly labeled with fluorochromes and optical sectioning using laser-scanning confocal microscopy.
Thompson
C T
CT
Department of Laboratory Medicine, University of California, San Francisco 94143-0808.
LeBoit
P E
PE
Nederlof
P M
PM
Gray
J W
JW
D002878:Chromosomes, Human, Pair 1 / Q000378:metabolism*; D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; D005557:Formaldehyde; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008545:Melanoma / Q000378:metabolism*; D008853:Microscopy; D008867:Microtomy; D016612:Paraffin Embedding; D012878:Skin Neoplasms / Q000378:metabolism*; D016707:Tissue Fixation
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; C069632:fluorescein-11-deoxyuridine triphosphate; D005557:Formaldehyde