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2. Fixing Agents, Stains and Mounting Media, , , , Fixing Agents and Preservatives, 1. Carnoy’s fluid, , 100% ethyl alcohol 7 30 cc, Glacial acetic acid - See., Chloroform 7 1S cc, , It is used for root tips, anthers, etc. and is, preferred for its great penetrating power., , 2. Formalin-Aceto-Alcohol, 50% or 70% ethyl alcohol — 90 ce., Glacial acetic acid - Sc, Formalin - Sec, Cis popularly known as FAA and is a standard, universal fixative. It is the most extensively used, fixing and killing agent,, 3. Formalin-Propiono-Alcohol, In the preparation of FAA use propionic acid, instead of acetic acid., 4, Randolph's modified Navashin fluid, Solution A. Chromic acid — Sem, Glacial acetic acid — 50 co, Distilled water — 320 ce, Solution B. Natural formalin — 200 cc, Saponin — 39m, Distilled water — 175 ce, , At the time of use, mix solutions A and B in, equal amounts. Recommended for buds, roots, tips and similar objects., , 5. Bouin's fluid, , Picric acid (1.5% aq. solution) — 75 cc., Formalin - Bee, Glacial acetic acid - Se, , This fixative is more useful than those with, chromic acid., Stains, 1. Acetocarmine, Dissolve 1 gm of stain in 100 ce of boiling 45%, acetic acid (or propionic acid). Cool and, , , , Append:, , employed (a) Heidenhain’s and (b) Delafied’s, , hematoxylin., , (a) Heidenhain’s hematoxylin,, Half per cent solution of the stain is, prepared in warm and distilled water. It is, then stored in dark in a closed bottle to, ripen for at least four days before use., , (b) Delafield’s hematoxylin., , (@ A saturated aqueous solution 4,, (100 cc) of ferric ~ammonium, sulphate is prepared., , Gi) One gram of stain is dissolved in 6 cc, of absolute alcohol., , (ii) Mixture of solutions 1 and 2 is, prepared. Add to this solution 25 cc, of glycerine and 25 cc of absolute, alcohol are added to this mixture., , (iv) The solution thus prepared is, , allowed to remain for sufficient time, till the colour becomes dark red., , decant, Add a few drops of saturated aqueous, solution of ferric acetate. Cool by keeping in, ice for at least twelve hours. Filter and store, the stock in refrigerator. For storage use, dropping bottle that is dark or covered with a, black paper., , 2. Aniline blue, (cotton blue, china blue, water blue), Aniline blue - 1 gm, Alcohol 90% or water - 100 ce, For better results stain or alcohol should be, slightly acidified with hydrochloric acid., 3. Crystal violet (Gentain violet), Crystal violet - 1 gm, Distilled water 100 cc, 4, Erythrosine, ‘The following are two recipes :, (a) Erythrosine = 1 gm, Alcohol 90% - 100 ce, (b) Erythrosine 1 gm, Absolute Alcohol - Sc, Clove oil - 95 ce, 5. Fast green, The following arestwo recipes :, (a) Fast green - 0.5 gm, Alcohol 90% - 100 ce, (b) Fast green - 05 gm, Absolute alcohol - 25 cc, Clove oil - 7S ce, 6. Gram’s iodine, Iodine - 2 gm, Potassium iodide (KI) - 3 ce, Distilled water - 300 ce, 7, Hematoxylin, It is a chromogen derived from logwood, Haematoxylon campechianum of Leguminosae., Two types of hematoxylins are commonly, [447, Safranin, (a) Safranin - gm, Alcohol 90% - 50 cc., Distilled water - 50 ce., (b) Safranin a 1 gm, Distilled water - 100 cc, , Mounting Media, , Glycerine jelly, , Soak some gelatin for 2 to 3 hours in cold, water, pour off the excess water and heat until, melted, To 1 part of this, add 1 5 parts of, glycerine and filter while still hot. Add 2 or 3, percent phenol. Still keeping the mixture hot, and fluid, add drop by drop a saturated, solution of methyl green in 50 percent alcohol,, until the glycerine becomes fully as dark as, green ink., , Lactophenol, , Mix equal parts of phenol crystals,, , glycerine and distilled water. Cott, , be mixed to stain fungi.
