Electron Microscopy Protocols--Kristen M.&nbs

Electron Microscopy Protocols
Credits

Part A: Tissue Preparation

LTP Physiology Protocol

Vibroslicer VT 1000S

Slice Dissection and Vibratoming in Agarose

Stoelting Tissue Chopper

Microwave Processing Protocol
Schedules:
a.    Standard Epon
b.    Epon/Spurrs
c.    Standard Epon with enhanced ribosomal
       staining

Routine Embedding of Tissue

Re-embedding Tissue

Tissue Embedding for Photography

Tissue Preparation Safety Protocols
a.    Protocol for Neutralizing the Aldehydes in our
       Fixative Waste
b.    Protocol for Disposing of Osmium Tetroxide
       Contaminated Plastic Containers (solid osmium
       tetroxide contaminated waste)

Part B: Microtomy

Glass Knife Making
Coating Grids
Denton Vacuum Carbon Evaporator
Photography of Epon Blocks using the Nikon Light Microscope
Hand Trimming Epon Blocks
Thick Sectioning
Thin Sectioning
Monitoring for Radioactivity
Staining Grids
Uranyl Acetate Recipe
Lead Citrate Recipe
Serial Sectioning
Ridge Pyramid Trimming for Serial Sectioning
Loading Serial Sections into Gimbols or Hex Rings

Part C: Electron Microscopy

Loading/Unloading Hex Rings into Gatan Rotational Holder
Loading/Unloading Gatan Rotational Holder into JEOL 2010
Loading/Unloading Gimbols into Rotational Holder
Loading/Unloading Rotational Holder into JEOL 1200 EX
Alignment Protocol for JEOL 1230
Photographing across the Depth of a Thin Section
Photographing Serial Sections
Unloading/Loading Film into the Electron Microscope
Developing Electron Microscope Film
Routine Scanning of EM Negatives Using Microtek Artix Scan 1800f Scanner
Scanning EM Negatives Using Polaroid SprintScan 45

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Electron Microscopy Protocols--Kristen M.&nbs