Chapter 7: Microsomes
7.2 Em observations of microsomes
Figure 7.3 Isolated microsomes
Figure 7.4 TEM of hepatocyte
- 1% Glutaraldehye (GTA)
- 1% Osmium tetroxide
- Epoxy or vinyl resin for TEM
- TEM photomicrograph of liver cells
- Transmission electron microscope
- Place a mm piece of the final pellet from
Exercise 7.1 into a small vial containing 1% GTA. Fix the pellet
for 2 hours.
- Rinse the pellet with water and place in three changes of
water for 30 minutes each, to remove any residual GTA.
- Post fix the lysosome pellet in 1% osmium tetroxide for 1
hour. Wash thoroughly with 3 changes of water (30 minutes each).
- Dehydrate the tissue by passing through a series of graded
alcohols or acetone, and embed in plastic blocks.
- Section the plastic blocks and place on coated grid. Double
stain with uranyl acetate and lead nitrate and examine with a
transmission electron microscope.
- Compare the view of the compacted lysosome pellet to the
structure and distribution of microsomes in an intact hepatocyte
(liver cell). Use figures 7.3 and 7.4 for reference.
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Cell Biology Laboratory Manual