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Protocols
Credits
Part A: Tissue Preparation
- Intravascular Perfusion
a. Via Thoracotomy
b. Via Heart Puncture
c. Via Tailvein
- LTP Physiology Protocol
- Acquiring Tissue Region
a. Vibroslicer VT 1000S
b. Slice Dissection and Vibratoming in Agarose
c. Stoelting Tissue Chopper
- Tissue Processing
a. Longhand
b. Microwave Enhanced Processing Schedules:
i. Standard Epon
ii. Epon/Spurrs
iii. Standard Epon with
enhanced ribosomal
staining
c. Embedding of Tissue
i. Routine Embedding of Tissue
ii. Re-embedding Tissue
iii. resurfacing block
- Tissue
Preparation Safety Protocols
a.
Neutralizing the Aldehydes
b.
Disposing Osmium Tetroxide Contaminated Plastic Containers
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
-
JEOL 1230
a. Loading/Unloading Gatan Holder
b. Alignment for JEOL 1230
- Loading/Unloading
Gimbols into Rotational Holder
- Thin Section
- Photographic Protocols
a. Film
i.
Film into the Electron Microscope
ii. Developing Electron Microscope Film
iii.
Routine Scanning of EM Negatives Using Microtek Artix Scan 1800f Scanner
iv. Scanning
EM Negatives Using Polaroid SprintScan 45
b. Digital
- Photographing
Serial Sections
- Other Electron Microscopes
a. Loading/Unloading
Gatan Rotational Holder into JEOL 2010
b. Loading/Unloading
Rotational Holder into JEOL 1200 EX
Part D: Maintenance of Electron Microscopy Equipment
Filament Replacement on the JEOL 1230 TEM
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