TEM Visualization of Microtubules
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TEM Visualization of Microtubules
LEVEL II
Materials
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Coated grid for TEM
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0.1 M ammonium acetate
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5% ethanol saturated uranyl acetate
- Transmission electron microscope
Procedure
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At the conclusion of a 37° C incubation from Exercise 9.6 , remove one drop (approximately 20 microliters) of the extract, place it on a collodion or formvar and carbon-coated EM grid, and allow it to remain for about 20-30 seconds. 6
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Rinse the grid carefully with 0.1 M ammonium acetate, and float the grid upside down on a drop of 5% uranyl acetate for 15 minutes. Rinse the grid with water, blot all excess fluids from the grid, and allow it to air dry.
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Examine the stained grid in a transmission electron microscope.
- Draw or photograph your observations -- calculate all appropriate dimensions (length, diameter) of the formed tubules.