Histopathology

                                ALZARIN RED Principle This method is based on the ability of certain dyes (alizarin red) to form lakes with calcium salts. Solutions                                     Alizarin red                  -           1gms                                     Distilled water             -       ...

                        ALCIAN BLUE — PAS Principle Alcian blue Solutions of varying PH are used to separate and identify the different acid mucins. Solutions  1) Alcian blue - 1 gm PH-02 10% Sulphuric acid     -       100ml PH-0.5  0.2 M Hydrochloric acid      -     100ml PH-10 0.1 M Hydrochloric Acid          -       10Oml PH-2.5 3% Acetic acid                        -        100ml PH-3.2 0.5%o Acetic acid    -           100ml. 2).5% Periodic acid  Schiff reagent Haematoxylin ...

                                                             ACID FAST STAIN Principle This organism has a fatty capsule which influences the penetration and resistance to removal of stain by acid and alcohol. Phenolic acid and heat are used to reduce surface tension , increasing the porosity and forcing dyes to penetrate this capsule . Solutions  Strong carbol fuchsin                                     Basic fuchsin              -           1 gm             ...

                                                WARTHIN STARRY STAT Clean all glassware in potassium dichromate sulphuric acid solution. Wash in tap water and rinse several times in triple distilled water. Solutions  1) Acidulated water                    Triple distilled water          -       1000 CC. Citric acid (1% or less) quantity sufficient to bring solution to a pH of 3.8-4.4 2) 1% Silver nitrate for impregnation. 3) 2% Silver nitrate solution (developer) 4) Gelatin solution                                 ...

                                      WADE FITE TECHNIQUE Principle   These organisms posses a capsule containing long chain fatty acid, (mycolic acid), which influences the penetration and resistance to removal of stain by acid and alcohol. The avoidance of defatting agents such as alcohol and Xvlene in the method are an attempt to conserve its fragile fatty capsule. Solution Carbol fuchsin ( Pink color) Strong Basic fuchsin                         -           1 gm Absolute Alcohol                               -       ...

                                                             VONKOSSA Principle This method is based on the conversion calcium salt in the salt of another metal ( Silver ) which itself is opaque.. Solutions A) Citrate buffer pH- 4.5            0.2M Disodium hydrogen phosphate     -        283 gms           Distilled Water                                              -        100ml B ) 0.1M Citric acid ...

       VERHOEFF VANGEISON STAIN Principle The disulphide bridges in the elastic fibers are converted to anionic sulphonic acid derivatives by treatment with iodine. These derivatives are strongly basophilic and capable of selective reaction with the basic dye (Haematoxylin). Solutions Solution A Haematoxylin              -           5gm Absolute alcohol          -           100ml Solution B Ferric Chloride            -           10gM Distilled water             -           100 ml Solution C Iodine    ...

                        VANGEISON STAIN Principle The method depends upon thę of tissue and the size of the dye molecule . Thus collagen Whish is relatively permeable takes red colour with acid fuchsin and muscle which in far less permeable takes yellow with picric acid . Weigert’s haematoxylin which is mordanted to an iron salt avidity to withstand treatment  with picric acid . which effectively removes alum haematoxylin from nclei. Solutions 1.        Weigert’s haematoxylin Solution A  Haematoxylin                       -           1gm Absolute Alcohol                   -        ...

                                                TOLUIDINE BLUE Solution Toluidine blue                -           1 ցո l  Distilled water               -           100Ml Procedure 1) Deparaffinise the section in Xylene for 10 mts  2) Bring the section to Water through descending series of alcohol. 3) Stain with methyl violet for 3-5 mts 4) Section should be mounted in aqueous media. 5) Section is kept in water and examined in mediately under microscope. Result Amyloid is distinguished by its dark red polarization colour.

SU DAN III OIL RED-O Lipids that exists as fats, have an affinity for the Sudan dyes. The adsorbing power of the fats is related to the dye concentration, temperature and the physical state of the fats. – Sections Cryostat sections are used for staining, since the fat would dissolve in the paraffin block sections. Solutions 1) Saturated solution of Sudan III, oil red        -        O in 70% alcohol. 2) Haematoxylin Procedure - Take 2 sections. 1)    Air dry sections and rinse in 70% alcohol. 2)    Stain with Sudan fed soil red for 10-20 mts. 3)    Rinse in 70% alcohol. 4)    Wash in water. Stain one section with Haematoxylin for 30 secs. 5)    Rinse in water and mount in glycerine jelly. 6)    Seal the edges of the coverslip with either molten wax or nail polish. Result       ...