Topic 6 - Preparation of tissues for electron microscopy

Question 1

Preparation of tissues for EM

Answer 1

1. Fixation (immersion or perfusion)
2. Staining/contrasting: osmium tetroxide
3. Dehydration: series with ethanol - and staining/contrasting: 70%ethanol with uranyl acetate
4. Intermediate solvent: propylene oxide
5. Embedding: synthetic resins
6. Preparation of semithin and ultrathin sections
7. Staining/contrasting: lead citrate

Question 2

Pre-embedding immunicytochemistry

Answer 2

triton X-100 is not allaowed to use
nitrogen helps the penetration of antibodies
the immunoreaction is carried out on 60-80 mm vibratome sections