[求救] Golgi staining問題
請問版上有人曾經做過Golgi staining這種染色法嗎？
最近實驗需要看神經走向所以想利用此染色來看
我有試染過但是很容易會連"微血管"也一起染到XD
不然就是染不太出來...cotrex的還OK...但是hippocampus就染不好了...
不知道有沒有人可以經驗分享的說...
有的話...非常感激 ^^"
我主要是採用網路上IHC WORLD的方法...方法如下：
http://www.ihcworld.com/_protocols/special_stains/golgi.htm
Solutions and Reagents:
Fixative Solution:
3% potassium bichromate ---------- 60 ml
10% formalin -------------------------- 20 ml
Mix well and store at room temperature.
2% Silver Nitrate Solution:
Silver nitrate ---------------------------- 2 g
Distilled water -------------------------- 100 ml
Mix well and store at room temperature.
Procedure:
1. Fix tissue blocks (less than 1 cm thick) in 10% formalin for 24 hours or
more. The alternatively you can also use the above fixative solution:
2. Place tissue blocks in 3% potassium bichromate for 3-7 days. Avoid light.
Change fresh solution each day.
3. Transfer tissue blocks into 2% silver nitrate solution for 1-3 day at room
temperature. Avoid light. (Note: before put blocks into silver nitrate
solution, use filter paper to absorb access solution. Change silver nitrate
solution (less than regular use) several times until brown participates do
not appear).
4. Cut vibratome sections at 60 um thick into distilled water.
5. Mount sections on to Superfrost Plus slides. Air dry for 10 minutes.
Dehydrate through 95% Alcohol, 100% alcohol, clear in xylene and coverslip.
Results:
Cortical neurons and processes ----------- black
Positive Controls:
Brain (Cortex).
謝謝囉～
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※ 編輯: mogi 來自: 140.109.42.116 (10/29 18:02)