Abstract

The primary focus in our laboratory is the study of higher order chromatin structure using three dimensional electron microscope tomography. Three dimensional tomography involves the deconstruction of an object by combining multiple projection views of the object at different tilt angles, image intensities are not always accurate representations of the projected object mass density, due to the effects of electron-specimen interactions and microscope lens aberrations. Therefore, an understanding of the mechanism of image formation is important for interpreting the images. The image formation for thick biological specimens has been analyzed by using both energy filtering and Ewald sphere constructions. Surprisingly, there is a significant amount of coherent transfer for our thick specimens. The relative amount of coherent transfer is correlated with the relative proportion of elastically scattered electrons using electron energy loss spectoscopy and imaging techniques.Electron-specimen interactions include single and multiple, elastic and inelastic scattering. Multiple and inelastic scattering events give rise to nonlinear imaging effects which complicates the interpretation of collected images.

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