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The Laboratory for Electron Microscopic Image Analysis V. J. Massari, PhD
Areas of Current Expertise <ul><li>Ultrastructure of the CNS and Peripheral Nerves </li></ul><ul><li>Immunocytochemistry <...
Advantages of Electron Microscopy in Neuroscience <ul><li>Resolution: </li></ul><ul><li>The resolving power of the E.M. is...
Disadvantages of Electron Microscopy <ul><li>It is difficult to see the forest because of the trees. </li></ul><ul><li>Sam...
Shared Resources are partially supported by the Georgetown Howard University Center for Clinical and Translational Studies...
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Electron Microscopic Image Analysis

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Electron Microscopic Image Analysis

  1. 1. The Laboratory for Electron Microscopic Image Analysis V. J. Massari, PhD
  2. 2. Areas of Current Expertise <ul><li>Ultrastructure of the CNS and Peripheral Nerves </li></ul><ul><li>Immunocytochemistry </li></ul><ul><li>Histochemistry </li></ul><ul><li>Definition of Neural Circuits using: </li></ul><ul><li>Anterograde Transport of Neural Tracers (Biotin) </li></ul><ul><li>Retrograde Transport of Neural Tracers (CTB) </li></ul><ul><li>Trans-synaptic Transport (Pseudorabies Virus) </li></ul><ul><li>Immunocytochemical labeling of neuro-transmitters and their receptors. </li></ul><ul><li>Preparation of tissues for EM. </li></ul>
  3. 3. Advantages of Electron Microscopy in Neuroscience <ul><li>Resolution: </li></ul><ul><li>The resolving power of the E.M. is much greater than that of the light microscope, which permits the visualization of sub-cellular structure. With the JEOL 1210, structure can be resolved at greater than 200,000X </li></ul><ul><li>This permits the visualization of both afferent and efferent elements of synapses and the precise definition of neuronal circuits. Circuits can be defined, for example, as axo-somatic, axo-dendritic, axo-axonic, myelinated, unmyelinated, etc. </li></ul>
  4. 4. Disadvantages of Electron Microscopy <ul><li>It is difficult to see the forest because of the trees. </li></ul><ul><li>Sampling area is very limited. </li></ul><ul><li>Requires preliminary survey with light microscopy. </li></ul><ul><li>Each labeled CNS element requires a different electron dense, black and white marker (DAB, TMB, Colloidal gold, Viruses). NO COLORS, correspondingly the state-of-the-art is only 2 elements labeled simultaneously. </li></ul><ul><li>Normal ultrastructure is compromised by immunocytochemical or histochemical labeling, making correct identification of structure difficult. </li></ul>
  5. 5. Shared Resources are partially supported by the Georgetown Howard University Center for Clinical and Translational Studies (GHUCCTS) 1UL1RR031975-01    In your publications, remember to mention support provided from the GHUCCTS.

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