Test Catageory
فئة التحليل
Histopathology
Refrence Books
كتب مراجع
1. Bancroft’s Histopathological Techniques
Skills Required
المهارات المطلوبة
1. Histology
2. Staining
Video Resources
مصادر فيديو
Aim
To demonstration of reticular Fibers.
Principle
The reticular Fibers is oxidized, and then sensitized with the iron alum, Which is replaced with silver. The silver is reduced with formalin to its visible Metallic state (black colour).
Solutions
1. Silver solution…………………………………stain reticular Fibers
To 5 ml of 10% aqueous silver nitrate add strong ammonia drop by drop until the
Precipitate which first forms is just dissolved. Add 5 ml of 3% sodium hydroxide
Then add strong ammonia drop by drop until the resulting precipitate is almost
Completely dissolved. Make up to 50 ml with distilled water
2-Potassium Permanganate…………………………………….oxidized the hexose
Sugar of reticular Fiber to aldehydes.
3-1% oxalic acid…………………………………………………bleaching agent
4- 2.5% aqueous iron alum………………………………………sensitizing agent
Iron alum (Ferric Ammonium Sulphate) ……………………..…. 2.5 g
Distilled water …………………………………………………..100 ml
5-10% formalin…………………………………………………..reducing agent
6-5% sodium thiosulfate………………………………………….fix the stain
7-Eosin ………………………………………………………….counter stain
Sample Type
he Gordon and Sweet’s technique can be used on deparaffinized and rehydrated tissue sections (4-5 microns) on positively charged slides. The preferred fixative is 10% neutral buffered formalin (NBF). Control samples can include normal liver.
Procedure
1. Deparaffinize sections with xylene then take through alcohols to water.
2. Oxidize in acidified potassium permanganate for 5 minutes
3. Rinse in distilled water.
4. Decolorize with 2% oxalic acid until the brown colour is removed
5. Rinse in distilled water.
6. Sensitize in iron alum for 10 minutes
7. Rinse in distilled water.
8. Impregnate in ammoniacal silver solution for 11 seconds
9. Rinse quickly in distilled water.
10. Immediately reduce with 10% aqueous formalin for 2 minutes
11. Wash in running tap water for 2 minutes
12. Rinse in distilled water.
13. Fix with 5% aqueous sodium thiosulphate (hypo) for 2 minutes
14. Wash in water for 2 minutes
16. Counter stain with Eosin for 2 minutes
17. Dehydrate, clear and mount
Results
Reticular Fiber…………………………………………………………………Black
Others………………………………………………………………….……. Red
Notes:
Diagnostic values:
Differential diagnosis of certain type of tumors.
Demonstration of abnormal pattern of reticular Fiber in liver diseases such as Liver cirrhosis, hepatocellular fibrosis.
Soha Alnoor
suha@mlsgaate.com
NEVER GIVE UP
ALWAYS HUSTLE FOR YOUR DREAMS
