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Histopathology Recipe Formulary
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Jenner Stain
Kaiserling's Fixative
Kopeloff & Beerman
Lactophenol
Lacto-Fuchsin
Leishman
Light Green
Light Green in Clove Oil
Lissamine Fast Red
Loeffler's Flagella Mordant
Loeffler's Flagella Stain
Loeffler's Meth Blue Lugols Iodine
Luxol Fast Blue-Cresyl Fast Violet
MacConkeys Capsule Stain
McFaydaen's Meth Blue
Mallory's Aniline Blue-Orange
Mallory's PTH
Mallory Heidenhain
Mann's Stain
Mann's Diff Alc
Masson Stain
May-Grunwald Stain
Methyl Green Orange G
Methyk Green Pyronin
Meth Green Pyronin (Pappenheim)
Methyl Violet 6B
Methyl Violet (Kopelhoff)
Methylene Blue-Fuchsin
Mucicarmine
Mullers Fixative
Muir's Capsule Stain
Neisser Meth Blue
Neisser Bismark Brown
Neisser Crystal Violet
Neisser Chrysoidin Neuroglia Mordant
Neutral Red
Neutral Red Light Green
Newman Stain (Modified)
Nigrosine
Nile Blue Sulphate
Nuclear Fast Red
Oil Red 4B
Oil Red O
Orange Fuchsin
Orcein Unna
Pal's Solution
Panchrome Stain
Papanicoleau
Pap OG6
Pasini Stain
Phloroglucinol
Phloxine

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JENNER STAIN - (Gurr)
Jenner stain 0.5 g
Pure methanol 100 ml
Dissolve by stirring.
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KAISERLING'S FIXATIVE - (Gurr)
Potassium acetate 100 g 
Glycerol 200 ml
Water 1000 ml
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KOPELOFF AND BEERMAN STAIN (see Gram) - (Gurr)
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LACTOPHENOL - (Gurr)
Lactic acid 25 ml 
Glycerol 50 ml
Distilled water 25 ml
Phenol, cryst 25 gm
Warm on a water bath until the phenol has gone into solution:  then store in a dark glass bottle.

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LACTO-FUCHSIN - (Gurr)
Acid fuchsin 0.1 g
Lactic acid 100 ml
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LEISHMAN STAIN - (Gurr)
Leishman stain 0.15 g 
Pure methanol 100 ml
May be dissolved by any of three methods: (a) place dye and alcohol in a flask, plug neck with cotton wool and warm in a water bath for 15 minutes with occasional shaking;  (b) triturate dye and methanol in a mortar for 15 minutes; (c) stirring cold for 2 hours.  The solution improves as a granule stain with keeping.

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LIGHT GREEN - (Masson)
Light green 2 g
Glacial acetic acid 2 ml
Water 100 ml
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LIGHT GREEN IN CLOVE OIL (or Cellosolve) - (Gurr)
Light green 1 g 
Clove oil or cellosolve 100 ml
Heat on water bath or electric oven or hotplate;  cool then filter.
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LISSAMINE FAST RED B (Picro-Lissamine) - (Lendrum)
Lissamine fast red B 1% aqueous 75 ml
Picric acid 1% aqueous 25 ml
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LOEFFLER'S FLAGELLA MORDANT - (Gurr)
Tannic acid 20% aqueous solution 100 ml 
Ferrous sulphate crystals 20 g
Basic fuchsin 10% in alcohol 10 ml
Water 40 ml
Dissolve iron salt in water by warming and add remainder;  do not filter.
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LOEFFLER'S FLAGELLA STAIN - (Gurr)
Basic fuchsin 10% in alcohol 20 ml
Aniline water 3% 80 ml
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LOEFFLER'S METHYLENE BLUE - (Gurr)
Methylene blue 0.5 g 
1% potassium hydroxide solution 1 ml
Ethanol, absolute 30 ml
Water 100 ml
Warm the water to about 50 degrees C., stir in methylene blue and add other ingredients; filter.
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LUGOL'S IODINE - (Gurr)
Iodine 1 g
Potassium iodide 2 g
Water 100 ml
Add iodine and iodide to about 25 ml water and when solution is complete add remainder of water.
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LUXOL FAST BLUE-CRESYL FAST VIOLET - (Kluver & Barrera)
Solution A  
Luxol Fast Blue 1 g
95% ethanol 1 litre
Acetic acid 10% 5 ml
Mix and filter.
Solution B
Cresyl Fast Violet 1 g
Water 1 litre
Before using add 5 drops 10% acetic acid to every 30 mls and filter.

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MacCONKEY'S CAPSULE STAIN - (Gurr)
Dahlia 0.5 g
Methyl green 1.5 g
Water 100 ml
Dissolve and add:
Basic fuchsin 10% alcohol solution 10 ml
Water 90 ml
Filter;  stand 2 weeks before use, keeps about 6 months

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McFADYEAN'S METHYLENE BLUE STAIN - (Gurr)
This is a 'ripened' Loeffler's methylene blue.  Loeffler's methylene blue is exposed to light and air for several months until it acquires a purplish tinge.

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MALLORY'S ANILINE BLUE-ORANGE (see Aniline Blue-Orange) - (Gurr)
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MALLORY'S PHOSPHOTUNGSTIC HAEMATOXYLIN - (Gurr)
Haematoxylin 1 g 
Water 800 ml
10% Phosphotungstic acid aq. soln. 200 ml
Dissolve haematoxylin in a little of the water by heating and mix.  Ripen by standing several weeks, or artificially by adding 12.5 ml. of a 1% solution of potassium permanganate.

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MALLORY-HEIDENHAIN'S (Rapid One Step Stain) - (Gurr)
Phosphotungstic acid 1 g
Orange G. 2 g
Aniline blue 1 g
Acid fuchsin 3 g
Distilled water 200 ml
Add ingredients individually to the water and stir to dissolve after each addition.
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MANN'S STAIN - (Gurr)
Methyl blue 1% aqueous solution 35 ml 
Eosin 1% 45 ml
Water 100 ml
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MANN'S DIFFERENTIATING ALCOHOL - (Gurr)
Ethanol, 70% 100 ml
Orange G. 7% aqueous 4 ml
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MASSON STAIN (see Ponceau Fuchsin) - (Gurr)
 
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MAY-GRUNWALD STAIN - (Gurr)
May-Grunwald stain 0.25 g
Pure methanol 100 ml
Dissolve by trituration;  shaking or stirring.
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METHYL GREEN-ORANGE G. (Kardos Stain) - (Gurr)
Kardos mixture 0.5 g 
Pure methanol 100 ml
Dissolve on an electric hot plate or by stirring.
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METHYL GREEN-PYRONINE - (Jordan & Baker)
(i)  Prepare a 0.5% aqueous methyl green solution and repeatedly extract with chloroform until chloroform extract is nearly colourless;  at least 8 extractions are needed.  
(ii)  Buffer Solution pH 4.8
Acetic Acid N/5
Sodium acetate M/t
81 ml
119 ml
(iii) Stain
Pyronine Y 0.5% aqueous

37 ml
Methyl green 0.5% (extracted as above) 13 ml
Buffer solution 50 ml
Solution (iii) keeps well for four months.
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METHYL GREEN-PYRONINE - (Pappenheim)
Methyl green 2% aqueous 4 ml 
Pyronine Y 5% aqueous 7 ml
Distilled water 100 ml
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METHYL VIOLET 6B - (Jensen)
Methyl violet 6B 5 g 
Water 1,000 ml
Dissolve and filter.
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METHYL VIOLET - (Kopelhoff & Beerman)
Methyl violet 6B (or crystal violet) 1% aqueous 30 ml 
Sodium bicarbonate 5% solution 8 ml
Should be freshly prepared every 3 or 4 days.
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METHYLENE BLUE-FUCHSIN - (Gray)
A. 1% aqueous methylene blue 50 ml 
Methanol, absolute 50 ml
B. 1% aqueous basic fuchsin 25 ml
Methanol, absolute 25 ml
Dissolve dyes and mix A with B.
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MUCICARMINE - (Gurr)
Mucicarmine 2.5 g 
Ethanol 50% 100 ml
Raise to boiling point in water bath, allow to boil 3 minutes, cool and filter.
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MULLER'S FIXATIVE - (Gurr)
Potass. dichromate 25 g 
Sodium sulphate 10 g
Water 1,000 ml
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MUIR'S CAPSULE STAIN MORDANT - (Gurr)
Saturated mercuric chloride solution (about 6%) 20 ml 
Tannic acid 20% solution 20 ml
Saturated potash. alum solution (about 10%) 50 ml
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NEISSER METHYLENE BLUE - (Gurr)
Methylene blue 2% aqueous solution  10 ml 
Ethanol, absolute 4 ml
Glac. acetic acid 10 ml
Water 180 ml
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NEISSER BISMARCK BROWN - (Gurr)
Bismarck brown 1 g 
Distilled water 500 ml
Heat water to about 60 degrees C., add dye and dissolve.
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NEISSER CRYSTAL VIOLET - (Gurr)
Crystal violet 1 g 
Ethanol, absolute 10 ml
Water 300 ml
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NEISSER CHRYSOIDIN - (Gurr)
Chrysoidin 1 g 
Water 300 ml
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NEUROGLIA MORDANT - (Anderson's)
1. Sodium sulphite 5 g 
Oxalic acid 2.5 g
Potassium iodide 5 g
Iodine 2.5 g
Distilled water 100 ml
Dissolve and add 5 ml glacial acetic acid
2. Ferric chloride (hydrated) 30% aqueous 5 ml
Water 95 ml
For use mix equal volumes of Nos. 1 and 2.
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NEUTRAL RED - (Jensen)
Neutral red 1 g 
Acetic acid 1% 2 ml
Water 1,000 ml
Dissolve and filter
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NEUTRAL RED-LIGHT GREEN - (Twort)
Neutral red-light green 1 g 
Clean sand 10 g
iso-Propyl alcohol 60% 100 ml
Grind all in a mortar for 5 minutes; filter.
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NEWMAN STAIN (Modified) - (Gurr)
Methylene blue 1 g 
95% ethanol 54 ml
Tetrachlorethane 40 ml
Glacial acetic acid 6 ml
Mix alcohol and tetrachlorethane, warm in water bath to 55 degrees C.  Add methylene blue and shake to dissolve;  cool and add acetic acid
Add basic fuchsin 1% in absolute ethanol 8 ml
Filter and keep well stoppered.
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NIGROSINE - (Gurr)
Nigrosine 5% aqueous solution.  
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NILE BLUE SULPHATE - (Gurr)
Nile blue 1 g 
Sulphuric acid pure 0.5 ml
Water 100 ml
Boil gently for 2 hours, occasionally making up volume with water;  cool, filter and make up to 100 mls, filter before use.
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NUCLEAR FAST RED - (Kernechtrot)
Nuclear fast red (Michrome No. 1022) 0.1 g 
Aluminium sulphate 5 g
Water 100 ml
Boil to dissolve.
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OIL RED 4B STOCK SOLUTION - (Gurr)
Prepare saturated solution as in the case of Oil Red O.  
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OIL RED O STOCK SOLUTION - (Gurr)
Oil Red O 0.5 g 
iso-Propyl alcohol 100 ml
Form saturated solution by warming in water bath or electric hot plate then filter
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ORANGE FUCHSIN - (Gurr)
Acid fuchsin 10% aqueous 10 ml 
Orange G, 3% aqueous 200 ml
Distilled water 30 ml
Absolute alcohol 60 ml
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ORCEIN - (Modified Unna)
Orcein 1 g 
Hydrochloric acid, pure 1 ml
Absolute ethanol 100 ml
Shake to dissolve, or stand overnight; filter.
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PAL'S SOLUTION - (Gurr)
Oxalic acid 1 g 
Sodium sulphite 1 g
Water 200 ml
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PANCHROME STAIN - (Pappenheim)
Methylene blue 1.0 g 
Toluidine blue 0.5 g
Azur 1 1.0 g
Methylene violet 0.5 g
Eosin Y 0.75 g
Pure methanol 250 ml
Glycerol 200 ml
Acetone 50 ml
Place all except in a flask and warm in a water bath or an electric hot plate for 20 minutes;  cool, add acetone, shake well and filter.
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PAPANICOLAOU STAIN - (EA36)
Eosin Y 10% aqueous solution 11.25 ml 
Light green SF 10% aqueous solution 2.25 ml
Bismarck brown 1% alcoholic solution 25 ml
Ethanol absolute 450 ml
Phosphotongstic acid 10% aq. solution 10 ml
Saturated aq. lithium carbonate 0.3 ml
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PAPANICOLAOU OG6 STAIN - (Gurr) 
Orange G satd. aqueous soln. 32.5 ml 
Phosphotungstic acid 10% aq. solution 0.75 ml
Ethanol 95% 475 ml
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PASINI STAIN - (Gurr)
Water blue-orcein 30 ml 
Eosin BA 2% in 50% alcohol 30 ml
Acid fuchsin 20% aqueous 4 ml
Glycerol 25 ml
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PHLOROGLUCINOL - (Gurr)
Phloroglucinol 1 g 
Water 100 ml
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PHLOXINE - (Lendrum)
Phloxine 0.5 g 
Calcium chloride 0.5 g
Water 100 ml
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PIANESE STAIN - (Gurr)
Malachite green 0.5 g 
Acid fuchsin 0.1 g
Martius yellow 0.01 g
Ethanol, absolute 50 ml
Water 150 ml
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PICRIC ALCOHOL - (Masson)
Picric acid saturated in 70% ethanol 60 ml 
Alcohol 95% 30 ml
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PICRO ANILINE BLUE - (Gurr)
Aniline blue water soluble 0.1 g 
Satd. aqueous picric acid 100 ml
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PICRO-FORMOL-ALCOHOL - ("Rossman's Fluid")
Picric acid satd. alcholic soln. 90 ml 
Formalin 10 ml
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PICRO FUCHSIN - (see Van Gieson)
 
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PICRO-PONCEAU S - (Gurr)
Ponceau S 1% aq. solution 10 ml 
Saturated aqueous picric acid  90 ml
Acetic acid 2% 2 ml
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POLYCHROME-METHYLENE BLUE - (Unna)
Methylene blue 1 g 
Potass. carbonate 1 g
Ethanol, absolute 20 ml
Water 100 ml
Dissolve methylene blue in water, add potass, carbonate and alcohol, place in water bath and evaporate to 100 ml
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POLYCHROME TOLUIDINE BLUE - (Martinotti)
Lithium carbonate 0.5 g 
Toluidine Blue 1.0 g
Water 75 ml
Glycerol 20 ml
95% ethanol 5 ml
Warm water, stir in lithium, then toluidine blue, dissolve and add glycerin-alcohol.
Ripen for one month.
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PONCEAU FUCHSIN - (Masson)
Acid fuchsin 0.3 g 
Ponceau de xylidine 0.7 g
G;acoa; acetic acid 1 ml
Water 100 ml
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PONDER'S STAIN - (Gurr)
Toluidine blue 0.1 g 
Acetic acid glacial 5 ml
Ethanol, absolute 10 ml
Water 485 ml
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PUGH'S STAIN - (Gurr)
Toluidine blue 1 g 
Acetic acid glacial 25 ml
Ethanol, absolute 10 ml
Water 465 ml
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RANDOLPH'S BLOOD DILUTING FLUID - (Gurr)
A. Methylene blue 0.1 g 
Propylene glycol 100 ml
Water 100 ml
B. Phloxine 0.1 g
Propylene glycol 100 ml
Water 100 ml
Mix equal volumes of A & B immediately before use; the mixture remains usable for 4 hours
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REGAUD'S FIXATIVE - (Gurr)
Potass. dichromate 3% solution 100 ml 
Formalin (neutralised by shaking with magnesium carbonate) 25 ml
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REGAUD'S HAEMATOXYLIN - (Gurr)
Haematoxylin 1 g 
Ethanol, absolute 10 ml
Glycerol 10 ml
Water 80 ml
Allow to ripen for 3 weeks, or, prepare with 10 ml. ripened 10% alcoholic haematoxylin instead of first two ingredients.
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RONGALITE WHITE - (Leuco Methylene Blue)
Methylene blue 0.5% aqueous 100 ml 
Hydrochloric acid pure 5 drops
Rongalite 3 g
Add acid to methylene blue solution, warm and add rongalite;  cool and filter.  Stable for a few days.
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SAFFRON - (Gurr)
Saffron 2 g 
Water 100 ml
Place in a flask and boil for an hour in a water bath.  Filter and add: 
Formalin 1 ml
Tannic acid 5% 1 ml
Note:  Saffron, whose, solution deteriorates after a few weeks, should not be confused with the cationic dye, safranin commonly used in histology.
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SAFRANIN ALCOHOLIC - (Gurr)
Safranin 1 g 
Water 50 ml
Dissolve and add:
Ethanol, absolute 50 ml
Filter.
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SAFRANIN AQUEOUS - (Gurr)
Safranin 1 g 
Water 100 ml

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