Atomic force microscopy and scanning near-field optical microscopy studies on the characterization of human metaphase chromosomes.
A better knowledge of biochemical and structural properties of human chromosomes is important for cytogenetic investigations and diagnostics. Fluorescence in situ hybridization (FISH) is a commonly used technique for the visualization of chromosomal details. Localizing specific gene probes by FISH combined with conventional fluorescence microscopy has reached its limit. Also, microdissecting DNA from G-banded human metaphase chromosomes by either a glass tip or by laser capture needs further improvement. By both atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM), local information from G-bands and chromosomal probes can be obtained. The final resolution allows a more precise localization compared to standard techniques, and the extraction of very small amounts of chromosomal DNA by the scanning probe is possible. Besides new strategies towards a better G-band and fluorescent probe detection, this study is focused on the combination of biochemical and nanomanipulation techniques which enable both nanodissection and nanoextraction of chromosomal DNA.
Oberringer
M
M
Department of Trauma-, Hand- and Reconstructive Surgery, University of Saarbrücken, 66421 Homburg, Germany. m.oberringer@mx.uni-saarland.de
Englisch
A
A
Heinz
B
B
Gao
H
H
Martin
T
T
Hartmann
U
U
D002469:Cell Separation / Q000379:methods; D002871:Chromosome Banding / Q000379:methods*; D002877:Chromosomes, Human / Q000648:ultrastructure*; D006801:Humans; D008214:Lymphocytes / Q000648:ultrastructure; D008677:Metaphase / Q000235:genetics; D042282:Microdissection / Q000379:methods*; D008846:Micromanipulation / Q000379:methods*; D018625:Microscopy, Atomic Force / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*