Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging |
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Affiliation: | 1. Department of Biology, College of William and Mary, Williamsburg, Virginia 23185, USA;2. Department of Biological Sciences, Hampton University, Hampton, Virginia 23668, USA |
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Abstract: | Integration of fluorescence light and transmission electron microscopy into the same device would represent an important advance in correlative microscopy, which traditionally involves two separate microscopes for imaging. To achieve such integration, the primary technical challenge that must be solved regards how to arrange two objective lenses used for light and electron microscopy in such a manner that they can properly focus on a single specimen. To address this issue, both lateral displacement of the specimen between two lenses and specimen rotation have been proposed. Such movement of the specimen allows sequential collection of two kinds of microscopic images of a single target, but prevents simultaneous imaging. This shortcoming has been made up by using a simple optical device, a reflection mirror. Here, we present an approach toward the versatile integration of fluorescence and electron microscopy for simultaneous imaging. The potential of simultaneous hybrid microscopy was demonstrated by fluorescence and electron sequential imaging of a fluorescent protein expressed in cells and cathodoluminescence imaging of fluorescent beads. |
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Keywords: | Transmission electron microscope Fluorescence microscope Cryo-microscopy Correlative microscopy Cathodoluminescence Fluorescent protein |
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