Brain Trigeminal Tissue Lab Report
Dissection Procedures: Brain, Trigeminal and Pterygopalatine Ganglia: After 48 hours, the brain was approached through the dorsal aspect of the skull. A T- shaped incision was given on the dorsal aspect of the head and the skin was reflected. The skull was nibbled avoiding damage to the brain. The brain was removed by releasing it gently from all its attachments from below and sides. Trigeminal ganglia were separated by cutting at the junction where the nerve enters into the brain stem. The brain was cleaned and washed with tap water followed by sterile water to remove excessive fixative materials. Olfactory bulbs were separated from the brain along with a small portion of the frontal pole. Cerebrum and cerebellum were also divided by a median incision into two
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III.5 Tissue Processing for Paraffin Embedding: Each tissue was then dehydrated in ascending grades of ethyl alcohol for 15- 30 minutes each (depending on size and type of tissue) separately:
50% alcohol → 70% alcohol → 90% alcohol → absolute alcohol I & II→ Xylene I & II (clearing agent) → Xylene and Wax (1:1 ratio) ¬→ 100% wax for 1 hours at 580C (2 changes) → Embedding in wax, blocked, labeled and stored.
III.6 Microtomy: Rotary microtome was used in order to prepare paraffin sections (5 to 10 µm thick) thickness with ribbon formation. 3-4 sections length ribbon selected at 10 section interval, floated on the hot water bath at 500 centigrade for straightening and then picked up on the egg albumin (Mayer’s albumin) coated glass slides. The slides were allowed to dry in the warm plate at 400 C for 2 hours and stored.
III.7 Staining Protocol:
Deparaffinization: Slides with paraffin section were deparaffinized in xylene for 15 minutes (2-changes). Hydration did with descending grades of alcohol for 5 minutes each as
III.5 Tissue Processing for Paraffin Embedding: Each tissue was then dehydrated in ascending grades of ethyl alcohol for 15- 30 minutes each (depending on size and type of tissue) separately:
50% alcohol → 70% alcohol → 90% alcohol → absolute alcohol I & II→ Xylene I & II (clearing agent) → Xylene and Wax (1:1 ratio) ¬→ 100% wax for 1 hours at 580C (2 changes) → Embedding in wax, blocked, labeled and stored.
III.6 Microtomy: Rotary microtome was used in order to prepare paraffin sections (5 to 10 µm thick) thickness with ribbon formation. 3-4 sections length ribbon selected at 10 section interval, floated on the hot water bath at 500 centigrade for straightening and then picked up on the egg albumin (Mayer’s albumin) coated glass slides. The slides were allowed to dry in the warm plate at 400 C for 2 hours and stored.
III.7 Staining Protocol:
Deparaffinization: Slides with paraffin section were deparaffinized in xylene for 15 minutes (2-changes). Hydration did with descending grades of alcohol for 5 minutes each as