Many biological processes lie out of the visual reach of scientists. The benefits of high-resolution electron microscopy are often offset by disruptive sample preparation requirements. Light microscopy allows easier sample prep and observations of living cells, but it has limited resolution. By manipulating how light waves behave, however, biophysicists are expanding the limits of light microscopy, and one of the latest advances--the 4Pi microscope--provides never-before-seen views of cellular components, including structures within the nucleus.
In a paper published in the Proceedings of the National Academy of Sciences, Joerg Bewersdorf of the Institute for Molecular Biophysics at The Jackson Laboratory, with collaborators Brian Bennett of the UMass Medical School and Leica Microsystems and Kendall Knight of the UMass Medical School, used the 4Pi microscope to examine the cellular response to a type of severe damage to the genetic material, DNA double-strand breaks. Such breaks provoke a rapid and highly coordinated series of events to identify and repair the damage. The response is critical, and there is an increased risk for cancer, developmental abnormalities and immunological problems when components of the repair processes are defective.
Traditional microbiological and genetics techniques can shed light on the molecular pathways of repair, but they don't address the astonishingly complex three-dimensional structure of the genetic material in the nucleus. 4Pi microscopy allows researchers to actually see the response in three dimensions, at resolutions down to 100 nm. Therefore, the r