Atomic force microscopy and scanning near-field optical microscopy studies on the characterization of human metaphase chromosomes

Eur Biophys J. 2003 Nov;32(7):620-7. doi: 10.1007/s00249-003-0309-8. Epub 2003 Jul 9.

Abstract

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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Cell Separation / methods
  • Chromosome Banding / methods*
  • Chromosomes, Human / ultrastructure*
  • Humans
  • Lymphocytes / ultrastructure
  • Metaphase / genetics
  • Microdissection / methods*
  • Micromanipulation / methods*
  • Microscopy, Atomic Force / methods*
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence / methods*