Abstract
Microdissection in combination with reverse painting fluorescence in-situ hybridization (FISH) is a very effective method to identify breakpoints and rearrangements of derived chromosomes and reveal the chromosomal origin of marker chromosomes. We describe an innovation that allows a convenient, fast and safe isolation of microdissected fragments as currently available protocols. The microdissected chromosomes are harvested in a collection drop located in a movable micropipette adjusted to a second micromanipulator under microscopic observation. We used this technique to analyze several cytogenetic aberrations. In order to evaluate the efficiency of our microdissection procedure, we compared the results obtained with microdissection probes made from only one fragment with those obtained with more than six microdissected fragments. In all cases, the single- fragment microdissections were sufficient to provide probes.
Similar content being viewed by others
References
Bohlander SK, Espinosa III R, LeBeau MM, Rowley JD, Diaz MO (1992) A method for the rapid sequence-independent amplification of microdissected chromosomal material. Genomics 13: 1322-1324.
Carter NP, Ferguson-Smith MA, Perryman MT et al. (1992) Reverse chromosome painting: a method for rapid analysis of aberrant chromosomes in clinical cytogenetics. J Med Genet 29: 299-307.
Guan X-J, Trent JM, Meltzer PS (1993) Generation of band-specific painting probes from a single microdissected chromosome. Hum Mol Genet 2: 1117-1121.
Guan X-J, Cargile CB, Anzick SL et al. (1995) Chromosome microdissection identifies cryptic sites of DNA sequence amplification in human ovarian carcinoma. Cancer Res 55: 3380-3385.
Heng HHQ, Tsui L-C (1993) Modes of DAPI banding and simultaneous in situ hybridization. Chromosoma 102: 325-332.
ISCN (1995) In: Mittelman F, ed. An International System for Human Cytogenetic Nomenclature. Basel: S. Karger, 1995.
Meltzer PS, Guan X-Y, Burgess A, Trent JM (1992) Rapid generation of region specific probes by chromosome microdissection and their application. Nature Genet 1: 24-28.
Müller-Navia J, Nebel A, Schleiermacher E (1995) Complete and precise characterization of marker chromosomes by amplification of microdissection in prenatal diagnosis. Hum Genet 96: 661-667.
Ried T, Schroeck E, Ning Y, Wienberg J (1998) Chromosome painting: a useful art. Hum Mol Genet 7(10): 1619-1626.
Rubtsov N, Senger G, Kuzcera H et al. (1996) Interstitial deletion of chromosome 6q: precise definition of the breakpoints by microdissection, DNA amplification, and reverse painting. Hum Genet 97: 705-709.
Senger G, Lüdecke H-J, Horsthemke B, Claussen U (1990) Microdissection of banded human chromosomes. Hum Genet 84: 507-511.
Senger G, Chudoba I, Friedrich U, Tommerup N, Claussen U, Brøndum-Nielsen K (1997) Prenatal diagnosis of a half-cryptic translocation using chromosome-microdissection. Prenat Diagn 17: 369-374.
Speicher MR, Ballar SG, Ward DC (1996) Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nature Genet 12: 368-375.
Telenius H, Carter NP, Bebb CE, Nordenskjöld M, Ponder BAJ, Tunnacliff A (1992) Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer. Genomics 13: 718-725.
Thangavelu M, Pergament E, Epinosa III R, Bohlander SK (1994) Characterization of marker chromosomes by microdissection and fluorescence in situ hybridization. Prenat Diagn 14: 583-588.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Weimer, J., Kiechle, M., Senger, G. et al. An Easy and Reliable Procedure of Microdissection Technique for the Analysis of Chromosomal Breakpoints and Marker Chromosomes. Chromosome Res 7, 355–362 (1999). https://doi.org/10.1023/A:1009263913478
Issue Date:
DOI: https://doi.org/10.1023/A:1009263913478