Combination of fluorescence in situ hybridization and scanning force microscopy for the ultrastructural characterization of defined chromatin regions

W. Fritzsche, L. Takács, G. Vereb, J. Schlammadinger, T. M. Jovin

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5 Citations (Scopus)

Abstract

Although the internal arrangement of interphase chromatin is still a matter of conjecture, there exists a large body of evidence for the compartmentalization of chromosomal domains. A study based on combined scanning force and optical microscopy of supramolecular chromatin spreads produced by isotonic lysis of cells suspended in phosphate-buffered saline has been conducted. The ultrastructure of fluorescent labeled chromosomes was resolved with the topographical contrast provided by the scanning force microscope. Fluorescence in situ hybridization was used to label specific DNA sequences. The location of different pericentromeric chromosome regions was determined by fluorescence microscopy and correlated with scanning force microscope topography. Using a single DNA probe, discrimination between labeled chromosome pairs of an aneuploid cell was possible, based on the different intensities of fluorescence signals. The results show that the in situ hybridization technique with fluorescence labeling is compatible with scanning force microscopy. The combination of these methods can be used for the specific identification and lateral localization of DNA sequences in spread chromatin, opening the possibility for the ultrastructural characterization of defined genes in the scanning force microscope.

Original languageEnglish
Pages (from-to)1399-1404
Number of pages6
JournalJournal of Vacuum Science and Technology B: Microelectronics and Nanometer Structures
Volume14
Issue number2
Publication statusPublished - Mar 1996

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chromatin
Chromosomes
Atomic force microscopy
Microscopes
Fluorescence
DNA sequences
microscopy
chromosomes
Scanning
fluorescence
scanning
deoxyribonucleic acid
microscopes
Fluorescence microscopy
Labeling
Topography
Optical microscopy
Labels
Phosphates
DNA

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Physics and Astronomy (miscellaneous)
  • Surfaces and Interfaces

Cite this

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abstract = "Although the internal arrangement of interphase chromatin is still a matter of conjecture, there exists a large body of evidence for the compartmentalization of chromosomal domains. A study based on combined scanning force and optical microscopy of supramolecular chromatin spreads produced by isotonic lysis of cells suspended in phosphate-buffered saline has been conducted. The ultrastructure of fluorescent labeled chromosomes was resolved with the topographical contrast provided by the scanning force microscope. Fluorescence in situ hybridization was used to label specific DNA sequences. The location of different pericentromeric chromosome regions was determined by fluorescence microscopy and correlated with scanning force microscope topography. Using a single DNA probe, discrimination between labeled chromosome pairs of an aneuploid cell was possible, based on the different intensities of fluorescence signals. The results show that the in situ hybridization technique with fluorescence labeling is compatible with scanning force microscopy. The combination of these methods can be used for the specific identification and lateral localization of DNA sequences in spread chromatin, opening the possibility for the ultrastructural characterization of defined genes in the scanning force microscope.",
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AU - Fritzsche, W.

AU - Takács, L.

AU - Vereb, G.

AU - Schlammadinger, J.

AU - Jovin, T. M.

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