Skip to main content
Log in

Imaging and manipulating chromosomes with the atomic force microscope

  • Published:
Chromosome Research Aims and scope Submit manuscript

Abstract

Polytene chromosomes from the salivary gland cells of Drosophila melanogaster were examined by atomic force microscopy. The atomic force microscope (AFM) was capable of resolving chromosomal features down to the limits of the tip sharpness, about 500 Å for pyramidal-shaped tips. Resolution was increased to 300 Å by using electron beam deposited (EBD) tips with high aspect ratios. This significantly exceeds the resolution obtainable with conventional optical microscopes, but at the cost of compromising the structural integrity of the sample. A reasonable compromise was achieved by using oxide-sharpened tips. In this case high resolution was obtained without sample degradation, but when desired these tips were also capable of sample disintegration with increased scanning force and rate. Thus, oxide-sharpened tips were used to precisely dissect defined chromosomal regions to illustrate their potential use in genetic mapping efforts. This study illustrates the utility of the AFM in the characterization and manipulation of chromosomes and chromosomal DNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Binnig G, Quate CF, Gerber C (1986) Atomic force microscope.Phys Rev Lett 56: 930–933.

    Google Scholar 

  • Drake B, Prater CB, Weisenhorn ALet al. (1989) Imaging crystals, polymers, and processes in water with the atomic force microscope.Science 243: 1586–1589.

    Google Scholar 

  • Guckenberger R, Arce FT, Hillebrand A, Hartmann T (1994) Imaging of uncoated tobacco mosaic virus by scanning tunneling microscopy.J Vac Sci Tech B, in press.

  • Guckenberger R, Heim M, Cerc Get al. (1994) Scanning tunneling microscopy of insulators and biological specimens based on lateral conductivity of ultrathin water films.Science 266: 1538–1540.

    Google Scholar 

  • Hansma H, Vesenka J, Siegerist Cet al. (1992) Reproducible imaging and dissection of plasmid DNA under liquid with the atomic force microscope.Science 256: 1180–1184.

    Google Scholar 

  • Henderson E (1992) Imaging and nanodissection of individual supercoiled plasmids by atomic force microscopy.Nucleic Acids Res 20: 445–447.

    Google Scholar 

  • Henderson E (1994) Imaging of living cells by atomic force microscopy.Prog Surf Sci 46: 39–60.

    Google Scholar 

  • Hill RJ, Rudkin GT (1987) Polytene chromosomes: the status of the band-interband question.BioEssays 7: 35–40.

    Google Scholar 

  • Hoh JH, Hansma PK (1992) Atomic force microscopy for high resolution imaging in cell biology.Trends Cell Biol 2: 208–213.

    Google Scholar 

  • Keller D, Chih-Chung C (1992) Imaging steep, high structures by scanning force microscopy with electron beam deposited tips.Surf Sci 268: 333–339.

    Google Scholar 

  • Keller D, Deputy D, Alduino A, Luo K (1992) Sharp, verticalwalled tips for SFM imaging of steep or soft samples.Ultramicroscopy 42–44: 1481.

    Google Scholar 

  • Lal R, John SA (1994) Biological applications of atomic force microscopy.Am J Physiol 266: 1–21.

    Google Scholar 

  • Miller R, Vesenka J, Henderson E (1994) Tip reconstruction for the atomic force microscope.SIAM, in press.

  • Pardue ML (1986) In situ hybridization to DNA in chromosomes and nuclei. In: Roberts DB, ed.Drosophila: A Practical Approach. Oxford: IRL Press, pp 111–137.

    Google Scholar 

  • Pardue ML, Kedes LH, Weinberg ES, Birnsteil ML (1977) Localization of sequences coding for histone messenger RNA in the chromosomes ofDrosophila melanogaster.Chromosoma 63: 135–151.

    Google Scholar 

  • Putman CAJ, van der Werk KO, de Grooth BGet al. (1992a) Atomic force microscope with integrated optical microscope for biological applications.Rev Sci Instrum 63: 1914–1917.

    Google Scholar 

  • Putman CAJ, van der Werk K, de Grooth BGet al. (1992b) A new imaging mode in atomic force microscopy based on the error signal.SPIE 1639: 198–204.

    Google Scholar 

  • Radmacher M, Fritz M, Hansma HG, Hansma PK (1994) Direct observation of enzyme activity with the atomic force microscope.Science 265: 1577–1579.

    Google Scholar 

  • Rasch P, Weidemann U, Wienberg J, Heckl WM (1993) Analysis of banded human chromosomes andin situ hybridization patterns by scanning force microscopy.Proc Natl Acad Sci 90: 2509–2511.

    Google Scholar 

  • Semeshin VF, Zhimulev IV, Belyaeva ES (1979) Electron microscope autoradiographic study on transcriptional activity ofDrosophila melanogaster polytene chromosomes.Chromosoma 73: 163–177.

    Google Scholar 

  • Vesenka J, Hansma H, Siegerist Cet al. (1992) Scanning force microscopy of circular DNA and chromatin in air and propanol.SPIE 1639: 127–137.

    Google Scholar 

  • Vesenka J, Manne S, Gibberson R, Marsh T, Henderson E (1993) Colloidal gold particles as an incompressible atomic force microscope imaging standard for assessing the compressibility of biomolecules.Biophys J 65: 992–997.

    Google Scholar 

  • Vesenka J, Miller R, Henderson E (1994) Three-dimensional probe reconstruction for atomic force microscopy.Rev Sci Instrum 65: 1–3.

    Google Scholar 

  • Wimber DE, Stefensen DM (1970) Localization of the 5S RNA genes ofDrosophila chromosomes by RNA-DNA hybridization.Science 120: 639–645.

    Google Scholar 

  • Yang J, Mou J, Shao Z (1994) Molecular resolution atomic force microscopy of soluble proteins in solution.Biochim Biophys Acta 1199: 105–114.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Eric Henderson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jondle, D.M., Ambrosio, L., Vesenka, J. et al. Imaging and manipulating chromosomes with the atomic force microscope. Chromosome Res 3, 239–244 (1995). https://doi.org/10.1007/BF00713049

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00713049

Key words

Navigation