Benha University Faculty of Veterinary Medicine Department of Pathology

STAI I ! T"#$ I%U"& • It means coloring of the tissues to identify; 'outine Stain ($)"*& +, $emato-ylin $arri.s Alum hemato-ylin& Hematoxylin Absolute alcohol Potassium alum Distilled water Mercuric oxide Preparation& 1" Dissol#e the hematoxylin in alcohol with gentile heat! " Dissol#e alum in water with aid of heat! $" Mix two solutions in a large Pyrex %rlenmeyer flas& ' 000 ml in ca(acity)! *" +ring the mixture to boil as ra(idly then turn off the flame and add the mercuric oxide slowly to (re#ent frothy! 5" ,he solution becomes dar& (ur(le in color and then cools under the ta( water! -" ,he solution becomes ready for staining! ." It recommended that 5 ml of the acetic acid to be added to each 100 ml to increase nuclear clarity! /, "osin& 5 gm 50 ml 100 gm 1000 ml !5 gm

• It is a synthetic dye! A, A0uous eosin 123 4 solution& • Formula %osin water solution Distilled water •Dissol#e and added 5 gm 1000 ml ml of the glacial acetic acid

B, Alcoholic eosin + 3 solution& •Formula& %osin / Distilled water 1 gm 0 ml

•Dissol#e and added 00 ml of 15alcohol! 5or6ing solution2 eosin 1 3 solution to 00 3 alcohol ' 12 $) ,he added 0!5 ml of glacial acetic acid to each 100 ml of the resulting solution! Procedures of staining 7ith hemato-ylin and eosin 1" Put the (araffin sections in xylol for 5 minutes to dissol#e the (araffin! " ,ransfer them to absolute alcohol the to 15 3 alcohol 4 and then to .0 3 alcohol ' two minutes in each solution)! $" 5ash in distilled water! *" 6tain with hematoxylin for 10 7 0 minutes!

5" 5ash in running water for at least 10 minutes until the sections become blue! -" 8ounter stain in eosin 1 3 for $ minutes! ." 9inse #ery carefully in water then in .0 3 alcohol for $0 seconds in each solution ! ,his is due to eosin is ready remo#able with water at .0 3 alcohol! 0" Dehydration occurs in 10 3 alcohol then in absolute alcohol 'each one for 1 minute) ! 1" 8lear in two or three changes in xylol ' each solution for 5 minutes )! 10"+efore the solution dries 4 (ut a dro( of 8anada balsm on it 4 then co#er it carefully with a clean co#er slide ' a#oid the formation of air bubbles )! 11":eft the sections dry in o#en at *0 o8 o#er a night! ;!+! the nucleus stained blue while the cyto(lasm stain (in&! Special Stains

Special stains of Fat &
< < < < Sudan III & stains fat yellowish brown Sudan IV ( Scarlet red * & stains fat orange to red 8il 'ed 8 stain & stains fat bright red 8smic Acid ( 8smium tetrao-ide *& stains fat blac&

< Special stains of connective tissue&
<Van !ieson stain 2 stain collagen red
Muscle4 cornified e(ith 4 amyloid yellow

oMasson Trichrome & stain 8!, blue o #rossman Trichrome & stain 8!, green o'eticulin Stain & 6tain reticular fiber blac&

o Special stains of Amyloid&
o#ongo 'ed & stain amyloid (in& to red oMethyl Violet 9 Methyl green 9 crystal violet 9 thianin stain & 6tain
amyloid red and the surrounding blue4 #iolet or green!

oVan !ieson stain & stain amyloid yellow
collagen red

oSpecial stains of iron&
o Perl.s Prussian :lue & stain iron blue


Special stains of #alcium&
o Von ;ossa Stain & stain calcium blac& o Ali<arin red S & stain calcium orange"red o A<an stain & stain calcium red


Special Stains of Mucin
o Alcian :lue& 6tain mucin blue


Mucicarmine & stain mucin magenta or red

o PAS& stains mucin red o #olloidal iron (=AMP=*& stain mucin blue


Melanin stains
o Masson,Fontana stain 2 stains melanin blac&

$o7 to remove the formalin pigments& • Formalin pigment is a fine dar& brown or blac& crystalline (reci(itates (roduced by lea&ed hemoglobin! For removal of such pigment> 1" Deparafini<ation of the (araffin sections and then (lace them in either; Solution +& Ammonia water 0 3 Alcohol .0 3 =r solution / A>ueous solution of hydrogen (eroxide $ 350 ml Acetone Place the sections in solution thoroughly in running water! 50 ml for 5 710 minutes then wash ml 100 ml

Place in solution 1 for 0 7 -0 minutes then rinse with ta( water !