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Related Experiment Videos

Probing chromatin with the scanning force microscope

W Fritzsche1, A Schaper, T M Jovin

  • 1Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

Chromosoma
|July 1, 1994
PubMed
Summary
This summary is machine-generated.

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Scanning force microscopy (SFM) visualizes biological material topography without fixation. This study reveals chromatin

Area of Science:

  • Biophysics
  • Molecular Biology
  • Microscopy

Background:

  • Scanning Force Microscopy (SFM) offers high-resolution imaging of biological material topography at interfaces.
  • Conventional electron microscopy techniques often require fixation, contrast enhancement, and labeling, which can alter specimen structure.
  • Visualizing delicate biological structures like chromatin requires advanced preparation methods compatible with high-resolution imaging.

Purpose of the Study:

  • To adapt conventional electron microscopy specimen preparation techniques for SFM visualization of chromatin ultrastructure.
  • To delineate the 'beads-on-a-string' morphology of nucleosomal assemblies using SFM.
  • To investigate the structural plasticity of chromatin in response to varying ionic strengths.

Main Methods:

Related Experiment Videos

  • Adaptation of specimen preparation techniques from conventional electron microscopy for SFM.
  • Hypotonic lysis of chicken erythrocytes followed by air drying to prepare nucleoprotein filaments.
  • SFM imaging to analyze the topography and dimensions of nucleosomal structures.
  • Exposure of air-dried metaphase chromosome samples to solutions of varying ionic strengths to observe structural changes.

    • Main Results:

    • A beaded substructure of the nucleoprotein filament was successfully visualized, consistent with a 'beads-on-a-string' morphology.
    • Individual nucleosomes appeared as round protrusions with an average height of 4-6 nm.
    • The center-to-center distance between adjacent nucleosome cores along the filament axis peaked at approximately 30 nm.
    • Reversible changes in the three-dimensional structure of metaphase chromosomes were observed upon altering solution ionic strength.

    Conclusions:

    • SFM, combined with adapted EM preparation techniques, provides nanometer-resolution imaging of chromatin ultrastructure without fixation.
    • The 'beads-on-a-string' model of nucleosomal organization is well-supported by the observed morphology and dimensions.
    • Chromatin structure exhibits dynamic, reversible changes in response to environmental factors like ionic strength, highlighting its flexibility.