Chromosome structure investigated with the atomic-force microscope

Bart G. de Grooth; Constant A.J. Putman; Kees O. van der Werf; Niko F. van Hulst; Geeske van Oort; Jan Greve

doi:10.1117/12.58188

1 May 1992 Chromosome structure investigated with the atomic-force microscope

Bart G. de Grooth, Constant A.J. Putman, Kees O. van der Werf, Niko F. van Hulst, Geeske van Oort, Jan Greve

Proceedings Volume 1639, Scanning Probe Microscopies; (1992) https://doi.org/10.1117/12.58188
Event: OE/LASE '92, 1992, Los Angeles, CA, United States

Abstract

We have developed an atomic force microscope (AFM) with an integrated optical microscope. The optical microscope consists of an inverted epi-illumination system that yields images in reflection or fluorescence of the sample. With this system it is possible to quickly locate an object of interest. A high-resolution image of the object thus selected can then be obtained with the AFM that is built on top of the optical microscope. In addition, the combined microscopes enable a direct comparison between the optical image and the topography of the same object. The microscope is used to study the structure of metaphase chromosomes of eukaryotic cells. The topography of metaphase chromosomes reveal grooved structures that might indicate spiral structure of the chromatin. High resolution images reveal structures that can be ascribed to the end loops of the chromatin. The resolution of the AFM images was improved by using sharper tips obtained by carbon deposition on the Si₃N₄ cantilevers using a scanning electron microscope. Chromosomes which are treated to reveal the G- banding pattern in the optical microscope display a similar pattern when viewed with the AFM, as is shown by a direct comparison.

Citation Download Citation

Bart G. de Grooth, Constant A.J. Putman, Kees O. van der Werf, Niko F. van Hulst, Geeske van Oort, and Jan Greve "Chromosome structure investigated with the atomic-force microscope", Proc. SPIE 1639, Scanning Probe Microscopies, (1 May 1992); https://doi.org/10.1117/12.58188

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