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Electron Microscopy

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More in this Support Team
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Support Teams / Electron Microscopy
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What We Do

The Electron Microscopy Facility enables biological research at micro- and nano-scales and provides Janelia researchers a nexus for multidisciplinary collaboration in high-resolution imaging. We offer state-of-the-art instrumentation, services, and training in sample preparation, transmission  electron microscopy and image analysis.  With 5,000-square-feet of space, we’re set up to prepare samples using conventional wet chemistry, microwave techniques and high-pressure freezing/freeze substitution. Our skilled staff will assist researchers and train them in the science and practice of electron microscopy as well.

 

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Equipment

  • FEI Tecnai G2 20 TWIN electron microscope with LaB6 gun and Gatan Ultrascan 4k x 4k quad port digital camera
  • FEI Spirit BioTWIN electron microscope with LaB6 gun and Gatan Ultrascan 4k x 4k quad port digital camera
  • Zeiss Nvision 40 FIB SEM for 3D nanoscale high-resolution imaging (equipment maintained by EM facility and operated by Hess Lab)
  • Zeiss Xradia 510 Versa for submicron X-ray imaging (equipment maintained by EM facility and operated by the Fetter Lab)
  • Wohlwend HPF Compact 01 high-pressure freezer for freezing suitably-sized unfixed, uncryoprotected samples without significant ice crystal damage
  • Leica EM AFS2 automatic freeze-substitution system for processing samples prepared by high-pressure freezing or progressive lowering of temperature (PLT) methods
  • Built-in UV lamp for UV-catalyzed resin polymerization at low temperature
  • Leica UC6 ultramicrotomes for room temperature and cryo semi-thin and ultra-thin sectioning. Each microtome is positioned in a separate room with adequate bench space and ancillary specimen preparation equipment so as to be self-contained sectioning areas
  • Leica KMR2 knifemaker for making glass knives
  • EMS 100 Glow Discharge unit for making support films on hydrophilic grids
  • Denton Explorer 14 Auto high-vacuum deposition system or carbon and metal coating that’s equipped with a quartz thin film monitor and rotary stage
  • Pelco BioWave Pro microwave oven with ColdSpot, SteadyTemp, and vacuum chamber options for maximum microwave sample preparation flexibility

Services

  • Operation of microscopes and acquisition of images
  • Sample fixation and processing
  • Sample sectioning and staining for TEM
  • Development of protocols for special research needs
  • Training

 

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Optimizing Immunogold Electron Microscopy for Neuroscience
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Dr. Wei-Ping Li

Dr. Wei-Ping Li completed her Ph.D. in cell biology with Jürgen Roth, the “immunogold master” who pioneered the use of immunogold EM on thin sections (Sedwick 2013).  Following in the footsteps of her mentor, Dr. Li is Janelia’s immunogold expert, developing protocols that enable researchers to analyze connectivity between identified neurons at the EM level.

Double immunogold labeling in mouse brain is used to distinguish different cell types.  Protocol was optimized to preserve antigenicity and synaptic ultrastructure (arrows).

Neuron expressing a fluorescent protein engineered by the Looger Lab was reconstructed from immunogold labeled serial sections of the mouse brain. Synapses (red arrows) are readily identifiable.  Figure is modified from Viswanathan et al. (in press).
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