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
For selective demonstration of muscles, collagen fibers, fibrin, and erythrocytes
Principle
Collagen fibers have strong affinity for high molecular weight of dyes, while Muscle, RBCs, and fibrin have affinity for low molecular weight of dyes (according to the tissue permeability and dye size molecule).
Solutions
Van Gieson solution:
1. Saturated aqueous picric Acid. (Small dye molecule)
2. 1% acid Fuchsin. 1 gm in 100 ml D.W (Large dye molecule)
3. Take 10 ml of 1% acid fuchsin and add to 90 ml Picric acid.
Weigert’s hematoxylin (Nuclear stain)
A. 1% alcoholic hematoxylin
B. Iron solution:
30% aqueous ferric chloride………………………………4 ml
HCL (Conc)………………………..………………….….1 ml
D.W……………………………………………………….95 ml
Take equal parts of solution A and B and mix them immediately before use.
Sample Type
Van Gieson’s stain can be used on a variety of samples, including:
Deparaffinized and rehydrated tissue section: These are sections of tissue that have been embedded in paraffin, then removed from the paraffin and rehydrated3.
Positively charged slides: These are slides that have been treated to have a positive charge, which helps tissue sections adhere to the slide
Procedure
1-Bring section to water.
2-Stain nucleus with Weigert’s solution for (10-15) min
3-Wash in water for (7-10) min
4-Stain with V.G working solution for (3-5) min
5-Blot with Filter Paper
6-Dehydrate, Clear and Mount.
Results
Nucleus: Black color.
Collagen fiber: Red color.
R.B.Cs, Muscles: Yellow color.
Notes:
1. Washing in water after V.G should be avoided, the color balance being Impaired by this.
2. Nuclear staining should be before application of V.G.
3. Celloidin sections washed in D.W. after V.G solution.
4. Differentiation step after Weigert’s hematoxylin is not needed because picric Acid in Van Gieson solution does this.
Diagnostic values:
1-Demonstrate the increase of collagen in diseases.
2-Assessment of healing process
3-Liver Cirrhosis
4- Used to differentiate between collagen and smooth muscle in tumors.
Soha Alnoor
suha@mlsgaate.com
Take equal parts of solution A and B and mix them immediately before use.
