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 demonstration of Elastic Fibers
Principle
Elastic Fibers are highly cross-linked by disulfide bond which oxidize by Lugol, s Iodine to give anionic sulphuric acid derivatives which react with haematoxylin to Give black colour
Solutions
1-Verhoff, s haematoxylin………………………………………………stain elastic Fibers
A-5% alcoholic haematoxylin…………stain……………………………20ml
B-10% fecl3…………. mordant and oxidizing agent——8ml
C-Lugol, s iodine—–oxidize disulfide bond of Elastic Fibers—–8ml
1g—-iodine
2g—k iodide dissolve in 100 ml of D.W
2-Van Gieson—–counter stain
3-2% fecl3——differentiator
4-5% sodium thiosulfate——remove excess iodine
Sample Type
Verhoeff’s stain can be used on deparaffinized and rehydrated tissue sections (3-5 microns) on positively charged slides2. The preferred fixative is 10% neutral buffered formalin (NBF). Control samples can include aorta, artery, or skin.
Procedure
1-Bring section to water
2-stain with Verhoff working solution for 15 mins
3– Wash in water
4-Differentiate with 2% fecl3 with microscopic control until elastic Fibers appears black.
5– Wash in water
6-place section in sodium thiosulfate 1 minute
7-counter stain in Van Gieson 1 minute
8-Rinse in 95% alcohol
9– Dehydrate in absolute-clear in xylene –mount in D.P.X
Results
Elastic Fibers—-black
Nuclei——blue to black
Collagen Fibers——red
Notes:
Diagnostic applications:
Arteriosclerosis
Atrophy of elastic Fibers
Other methods:
-Aldehyde fuchsin
Soha Alnoor
suha@mlsgaate.com
Verhoeff’s Stain, also known as Verhoeff’s Elastic Stain (VEG) or Verhoeff–Van Gieson stain (VVG)
