PROBING CHROMATIN WITH THE SCANNING FORCE MICROSCOPE

With the scanning force microscope (SFM), one can image the topography of biological material adsorbed at air-solid or liquid-solid interfaces with up to nanometer resolution. In principle, fixation, contrast enhancement, and labeling are not required. We have adapted specimen preparation techniques of conventional electron microscopy for visualizing chromatin ultrastructures in the SFM. A beaded substructure of the nucleoprotein filament was obtained after hypotonic lysis of chicken erythrocytes and air drying. The beads-on-a-string morphology of the basic nucleosomal assembly was well delineated. The nucleosomes appeared as round protrusions with an apparent height of 4-6 nm. The histogram of center-to-center distances between adjacent nucleosome cores along the filament axis had a peak at similar to 30 nm. Reversible changes in the three-dimensional structure were observed upon exposure of air-dried samples of metaphase chromosomes to solutions of different ionic strengths.