School Science Lessons
Appendix B, Biology appendix
2014-09-15
Please send comments to: J.Elfick@uq.edu.au

1.0.0 Appendix B. Biology
Table of contents
9.1 Agar, media and nutrient solutions
4.0 Biology fixatives
1.0 Biology media and solutions
16.1 Biology supplies, (commercial websites)
6.0 Culture media for identification of fungi
18.7.1 Drinking water test media
9.0.0 Glues and pastes, adhesives, gums, copying fluid
1.14 Immersion oil
Indicators, "Prof Bunsen", (commercial website)
4.1 Insect fixing fluids
Microscopes, microscopes products, "Scientrific", (commercial website)
2.0 Microscopy adhesives
3.0 Microscopy stains
Plants, common names first
Plants, Dicotyledons
Plants, Family names, dicotyledons
Plants, Monocotyledons
7.0 Prepare acids and bases
8.0 Prepare salt solutions
39.0 Prepare standard buffer solutions

9.1 Agar, mediums and nutrient solutions
9.2.14 Basal agar medium
9.2.32 DRBC, (food spoilage medium)
18.7.1 Drinking water test media
9.2.16 Glucose nutrient agar
9.2.31 Glucose nutrient agar medium
9.2.17.0 Malt extract agar medium
9.2.30 Mannitol yeast extract agar, (MYEA)
9.2.19.2 Milk agar medium
9.2.18 Minimal agar medium
9.2.23 MS agar medium
9.2.27 Nitrogen-free mineral salts agar medium
9.2.19 Nutrient agar medium, starch, milk
9.2.33 Nutrient agar, Microbiological base ingredients
9.2.19.1 Starch nutrient agar medium
9.2.21 Urea agar medium

1.0 Biology media and solutions
1.1 Acid alcohol, biology solution
1.2 Alcohol, (ethanol), absolute alcohol, biology solution
9.2.24 BAP medium
9.2.15 Basal broth medium
1.8 Basal salt solutions, biology solutions
1.3 Benedict's reagent, Benedict's solution, biology solution
9.2.25 Buffer reagent, phosphate buffer reagent
1.4 Carbol xylol, biology solution
9.2.26 Domestos solution, NaClO
1.5 Fluorescein solution, biology solution
1.6.1 Gram's iodine solution
1.6 Iodine solution, biology solution
9.2.12 Liquid broth media
3.15 Lugol's iodine solution, microscopy stain
9.2.17.1 Malt extract broth medium
3.27 Methyl cellulose solution, methocel, (low substitution)
9.2.20 Nutrient broth medium
9.2.29 Nutrient gelatin
1.13 Phenylthiocarbamide, PTC, phenylthiourea, PTU
9.2.11 Sterile media or solutions
1.7 Ringer solution, biology solution
9.2.28 Salt solution
1.9 Scott's blueing solution, biology solution
1.10 Seawater substitute, biology solution
1.11 Sodium thiosulfate solution, biology solution
9.2.11 Sterile media or solutions
9.2.30 Tensides, alkylphenol ethoxylates
9.2.22 Vinegar bacteria medium
3.13.6 Weigert's haematoxylin, microscopy stain
1.12 Xylene and methylbenzoate, biology solution
9.0.0 Glues and pastes, adhesives, gums, copying fluid
1.3 Adhesives, List of adhesives
9.2.0 Commercial glues and pastes, (in plastic containers)
9.1.0 Flour and milk glues
9.1.1 Flour glue
9.1.2 Milk glue
3.100 Prepare casein plastic from milk
9.1.3 Wallpaper paste
9.1.4 Turpentine copying fluid

2.0 Microscopy adhesives
2.4 Canada balsam mounting medium
2.5 DPX mountant
2.1 Glycerine jelly, adhesive to stick sections to microscope slides
2.2 Haupt's adhesive, adhesive to stick sections to microscope slides
2.3 Meyer's albumen, adhesive to stick sections to microscope slides
2.6 Peru balsam mounting medium

3.0 Microscopy stains, histology stains
3.1 Acetic alcohol, microscopy stain
3.3.0 Aceto-carmine, microscopy stain
3.2 Aceto-orcein stain, microscopy stain
3.30 Alcian blue stain
3.3.1 Alizarin indicator
3.3.3 Alizarin red S indicator
3.0 Alizarin yellow R indicator
3.5.1 Aniline blue stain
3.4 Aniline hydrochloride, microscopy stain
3.5 Aniline sulfate, microscopy stain
3.5.2 Basic fuchsin, microscopy stain
3.5.4 Calberla's pollen stain
3.5.3 Carbol fuchsin, microscopy stain
3.6 Carmine stain, microscopy stain
10 Congo red, microscopy stain, acid-base indicator
3.13.1 Delafield's haematoxylin, microscopy stain
3.8 DCIP, (2,6-dichlorophenol-indophenol), microscopy stain
3.13.2 Ehrlich's haematoxylin, microscopy stain
3.10 Eosin, tetrabrornofluorescein, microscopy stain
3.28 Giemsa microscopy stain
3.11.0 Gram stain, microscopy stain
3.11.1 Crystal violet microscopy stain
3.34 Fluorescence staining of cells and tissues
3.13.0 Haematoxylin, C16H14O6, microscopy stain
3.13.3 Harris modified haematoxylin, microscopy stain
3.13.5 Heidenhain iron haematoxylin, microscopy stain
3.15 Lugol's iodine solution, microscopy stain
3.13.4 Mayer's haematoxylin, microscopy stain
3.16 Karo syrup mountant, microscopy stain
3.17 Lactophenol, microscopy stain
3.18 Lactophenol cotton blue, microscopy stain
3.19 Leishman's stain, microscopy stain for white blood cells, Wright's stain
3.19a Methylene blue, microscopy stain
13a Methyl violet, 2B, C24H28N3Cl, basic violet 1
13b Methyl violet 10B, C25H30N3Cl, basic violet 3
23.0 Neutral red, microscopy stain
3.31 Nuclear fast red stain
3.32 Oil red stain
3.21 Orange G, microscopy stain
3.33 Papanicolaou stain
3.22 Phloroglucinol, (10% solution), microscopy stain
16.1.6 Resazurin microscopy stain, test for milk
3.23 Safranin, microscopy stain
3.24 Schulze's solution, (chlor-zinc-iodine), microscopy stain
3.25 Toluidine blue, microscopy stain
3.13.6 Weigert's iron haematoxylin, microscopy stain
3.12 Ziehl–Neelsen stain

4.0 Biology fixatives
4.1 Aceto-alcohol, biology fixative
4.17 Bouin's solution, biology fixative
4.8 Carnoy's fluid, biology fixative
4.16 CRAF, (chromic acid, acetic acid, formalin), biology fixative, for plant material
4.18 Decalcifying solution
4.19 Differentiation solution
4.2 Ethanol solution
4.3 FAA, (formalin, acetic acid, alcohol), biology fixative, for plant material
4.4 Formaldehyde, formalin, biology fixative, for animal material
4.5 Formol-saline, biology fixative, for marine animals
4.20 Glutaraldehyde solution, microscopy fixative
4.6 Zenker's fluid, biology fixative, for animal material
4.1 Insect fixing fluids
4.15 Barber's relaxing fluid, insect-fixing fluid
4.9 KAA, insect-fixing fluid
4.10 Kahle's fluid, insect-fixing fluid
4.12 Lacto-alcohol, insect-fixing fluid
4.11 Oudeman's fluid, insect-fixing fluid
4.13 Pampl's fluid, insect-fixing fluid
4.14 Sugaring mixture, insect-fixing fluid

6.0 Culture media for identification of fungi
6.1 Calcofluor white with 10% KOH, identify fungi
6.2 Cellotape flag preparations, identify fungi
6.3 Cornmeal agar, identify fungi
6.4 Cornmeal glucose sucrose yeast extract agar, identify fungi
6.5 Czapek Dox Agar, identify fungi
6.0 Direct microscopic mounts or squash preparations
6.6 Indian ink mounts
6.7 Lactophenol cotton blue, (LPCB), identify fungi
6.13 Loeffler serum medium
6.14 MacConkey agar
6.12 Orcinol-Bial's Reagent
6.9 Potassium hydroxide with chlorazol black, identify fungi
6.10 Potato dextrose agar, identify fungi
6.11 Rice grain slopes, identify fungi

1.1 Acid alcohol, biology solution
Purchase: Acid Alcohol solution for microscopy
Acid alcohol is used for cleaning slides and coverslips and decolorizing some stains, e.g. haematoxylin.
Prepare acid alcohol solution: 100 mL 70% alcohol + 1 mL hydrochloric acid
1.2 Alcohol, (ethanol), biology solution
Purchase: Ethanol ACS reagent, 99.5% (200 proof), absolute
Ethanol is used in various concentrations for preserving and dehydrating. Absolute alcohol bottles must be stoppered at all times, since alcohol readily absorbs water from the atmosphere. Remove water from alcohol by using drying agents, e.g. anhydrous copper (II) sulfate.

1.3 Benedict's reagent, Benedict's solution
Purchase: Benedict's reagent for the quantitative determination of sugars
Prepare Benedict's solution:
1. Dissolve 173 g of sodium citrate and 100 g of anhydrous sodium carbonate in 800 mL of water then filter the solution. Dissolve 17.3 g of copper sulfate in 100 mL of water and add this to the filtered solution. Make up to 1 L with water.
2. Solution A: Dissolve with heat 173 g sodium citrate, 100 g anhydrous sodium carbonate, Na2CO3, in 800 mL water. Filter and dilute to 850 mL.
Solution B Dissolve 17.3 g copper sulfate crystals, CuSO4.5H2O in 100 mL water. Pour Solution B with stirring, into Solution A, and make up to 1 litre.
3. Solution A: Dissolve 17.3 g of sodium citrate and 10 g of anhydrous sodium carbonate in 60 mL of water.
Solution B: Dissolve 1.73 g of copper sulfate crystals in 20 mL of water. Pour solution B into solution A while stirring, and make up to 1 litre.

1.4 Carbol xylol, biology solution
Purchase: m-Xylol, m-Xylene, anhydrous, (1,3-Dimethylbenzene ), Metaxylene, C8H10
Xylol, Low cost: from hardware stores and paint stores, as xylenes, (mixed dimethylbenzene isomers)
Purchase: carbolic acid, phenol, hydroxybenzene, benzenol, C6H6O, C6H5OH
BE CAREFUL! Use gloves!
Carbol xylol solution , a mixture of 3 parts xylol and 1 part melted crystal carbolic acid is used for dehydrating in staining techniques.
Prepare carbol xylol solution: Mix 25 g carbolic acid, with 100 mL xylene, C8H10. The phenol dissolves slowly, and cools when dissolving.

1.5 Fluorescein solution
Purchase: Fluorescein (free acid), Dye content 95 %, C20H12O5
Prepare fluorescein solution: Dissolve one gram of fluorescein in 100 mL methylated spirit.

1.6 Iodine solution, biology solution
Purchase: Iodine, trace metals basis, I2
Purchase: Potassium iodide, ACS reagent, 99.0%, KI
Iodine solution is used for tests for starch, (Fehling's test)
Store iodine container inside a second container because the lid of iodine container may deteriorate. Iodine is scarcely soluble in water, so iodine solution is iodine dissolved in potassium iodide solution. Use safety glasses and nitrile chemical-resistant gloves when weighing solid iodine because it is harmful and corrosive. Once in solution the amounts of iodine used are minute. Specific tests for
starch giving blue-black colour and general stain. Kills and fixes living material and makes cytoplasm and nucleus more visible. Lignin walls in xylem stained brown, leaving cellulose walls of parenchyma relatively unstained. Stains proteins brown, nuclei dark brown, chloroplasts brown but black if starch present, cellulose cell walls yellow, lignified cell walls deep yellow to brown.
Prepare iodine solution:
1. Dissolve 2 g potassium iodide in water. Add 1 g iodine crystals.
2. Dissolve 1 g iodine and 4 g potassium iodide in 300 mL water.
3. Add l g iodine crystals and 5 g potassium iodide to 50 mL water. Dilute to 100 mL
4. Dissolve 10 g of potassium iodide in 100 mL of deionized water and add 5 g of iodine crystals.
5. Dissolve 15 g of potassium iodide in 20 mL of deionized and add 3 g of iodine crystals. Dilute to 1 litre.
5. Add iodine flakes to methylated spirit to form alcoholic iodine solution.

1.6.1 Gram's iodine solution
Purchase: Gram's iodine solution
Gram's iodine is used for an indicator for the presence of starch, detection of alkaloids, source of iodine for iodometric titrations, and source of iodine / iodide.
Prepare Gram's iodine solution: 2% iodine and 3% potassium iodide in 70% ethanol.

1.7 Ringer solution, biology solution
Ringer solution is a mixture of salts that dissolve in water to form a physiological saline solution. Prepare fresh before use.
Purchase: Ringers solution tablets for microbiology
Prepare Ringer solution:
1. 0.9 g sodium chloride, 0.042 g potassium chloride, 0.025 g calcium chloride, 100 mL deionized water.
2. 7.2 g sodium chloride, 0.37 g potassium chloride, 0.17 g calcium chloride, dissolve in deionized water, add deionized water to 1 litre, adjust pH to 7.3-7.4.
3. 2.25 g / litre sodium chloride, 0.105 g / litre potassium chloride, 0.12 g / litre calcium chloride.6H2O, 0.05 g / litre sodium bicarbonate. One tablet makes 500 mL of quarter strength Ringer solution. To prepare quarter strength Ringer Solution, dissolve 1 tablet in 500 mL distilled water. Sterilize by autoclaving at 121°C for 15 minutes.
4. Dissolve 1 Ringer tablet in 500 mL deionized water and then autoclave it.

1.8 Basal salt solutions, biology solution
Basal salt solutions are used as an irrigating, transporting and diluting fluid while maintaining intra-cellular and extra cellular osmotic balance. They provides cells with water and certain bulk inorganic ions essential for normal cell metabolism. They may be combined with a carbohydrate, such as glucose, provides the principle energy source for cell metabolism. Also, they provides a buffering system to maintain the medium within the physiological pH range (7.2-7.6).
Basal salt solutions for cell culture include the following:
Dulbecco's phosphate buffered saline contains no bicarbonate, so a pH shift during filtration is less likely.
Earle's balanced salts contains 2.2 gm / litre sodium bicarbonate, may require pH adjustment after filtration.
Hanks' balanced salts contains 0.35 gm / litre sodium bicarbonate, may require pH adjustment after filtration.
Prepare salt solution: Dissolve 9 g sodium chloride in 1 litre of deionized water.

1.9 Scott's blueing solution, biology solution
Purchase: Scott's Tap Water Substitute Concentrate, 10 ×
Scott's blueing solution is used with haematoxylin to develop blue colour.
Prepare Scott's blueing solution: 2 g sodium bicarbonate, 20 g magnesium sulfate, 1 litre deionized water.

1.10 Seawater substitute, biology solution
Prepare seawater substitute: Dissolve in 2 litres of water, 45.0 g sodium chloride, 3.5 g magnesium sulfate, 5.0 g magnesium chloride, 2.0 g potassium sulfate.

1.11 Sodium thiosulfate solution, biology solution
It is used to decolorize iodine and wash iodine from tissue.
Prepare sodium thiosulfate solution: Dissolve 5 g sodium thiosulfate, (hypo), in 100 mL deionized water. Add drops of iodine solution. The colour of iodine vanishes. Add drops of weak solutions of acidified bleaching fluid, (calcium hypochlorite), or citric acid or sulfuric acid. The colour of iodine solution returns.

1.12 Xylene and methyl benzoate, biology solution
Xylene, C8H10, C6H4(CH3)2, m-Xylene anhydrous, 1,3-dimethylbenzene, solvent, Xylene, Toxic by all routes, highly flammable
Methyl benzoate, C6H5COOCH3, solution almost colourless, fragrant liquid, Toxic if ingested, use < 50 mL or g per activity
They are used during staining procedures. These chemicals are highly flammable, toxic and readily absorbed through the skin. Care must be exercised when handling. These are not miscible with water.

1.13 Phenylthiocarbamide
Phenylthiocarbamide, PTC, phenylthiourea, PTU, N-Phenylthiourea, 1-Phenyl-2-thiourea, C7H8N2S, C6H5NHCSNH2
See 9.24.2: PTC tasters and non-tasters
Phenyl thiocarbamide is toxic but is safe at a concentration < 0.13%.
Prepare PTU solution: Weigh 0.13g of solid phenylthiocarbamide on a watch glass. Dissolve the solid in 100 mL deionized water. Cut strips of absorbent paper, 5 cm × 1 cm. Soak the strips in the solution, remove with forceps, drain and dry the papers on a drying tray in an incubator < 50oC. Store the indicator papers in a sealed bottle.
Prepare strips of paper soaked in solutions ×2 or ×10 as dilute after diluting the above solution 50 mL to 100 mL or 10 mL to 100 mL. Rinse the mouth after each tasting trial. The chemical has a very persistent bitter taste, so handle the strips of paper with forceps.

1.14 Immersion oil
Purchase: Immersion oil for microscopy, refractive index n20 / D 1.516, viscosity 100-120 mPa.s(20 °C), density 1.025 g / mL at 20 °C
Immersion oil, high refractive index, for finer resolution and brightness, (mercury e line, (5461 A) specification)
Immersion oil for microscopy, is used for high resolution (1000 ×) light microscopy work under an oil immersion objective lens.
By placing a drop of immersion oil with the same refractive index as glass between the cover slip and objective lens two refractive surfaces are eliminated, so that greater magnifications can be achieved while still preserving good resolution.

2.1 Glycerine jelly
Glycerine jelly, adhesive to stick sections to microscope slides
Glycerol Gelatin aqueous slide mounting medium, for histological use, must be warmed before use then returns to semi-solid state.
Prepare glycerine jelly: Soak 10 g gelatine in 60 mL water for 2 hours. Add 70 mL glycerine and 1 g phenol crystals. Heat the solution gently in a water bath and then cool. To soften the jelly before embedding heat in water bath.

2.2 Haupt's adhesive
Adhesive to stick sections to microscope slides
Purchase Haupt's Adhesive Concentrate
Prepare Haupt's adhesive: Dissolve 1 g gelatine in 100 mL water at 30oC. Add 2 g phenol crystals and 15 mL glycerine. Stir, cool and filter.

2.3 Meyer's albumen
Meyer's albumen, adhesive to stick sections to microscope slides
Purchase Albumin from bovine serum, lyophilized powder, 96% (agarose gel electrophoresis), culture-tested powder
Prepare Meyer's albumen: Beat an egg white until well broken up, but not stiff. Pour into a tall cylinder and leave to stand overnight. Add equal volume of glycerine to liquid collected from bottom. Add a thymol crystal to prevent growth of fungus.

2.4 Canada balsam mounting medium
Balsams are hard varnishes and fragrant perfumes. They have oleoresins containing volatile essential oils and nonvolatile resins. Canada balsam, Canada turpentine oil, balsam of fir, American silver fir, balm of gilead fir, is a natural turpentine used in microscopy because the refractive index is similar to the refractive index of glass, (n = 1.55). It comes from the thin resin in bark of the balsam fir tree, Abies balsamea, (Pinus balsamea), Pinaceae. It is used as a stable apolar hydrophobic mounting medium for light microscopy slide preparation, especially when long term storage is desired. It was used to conserve microscope samples, but nowadays synthetic resins are used.
Purchase: Canada balsam, Mounting medium for microscopy, Balsam Canada

2.5 DPX mountant
Purchase: DPX Mountant for histology
DPX is a neutral synthetic mountant for histology, it is a mixture of distyrene, plasticizer, and xylene, this synthetic resin mounting medium can replace xylene-balsam, dries quickly and preserves stain, suitable for haematoxylin eosin staining.
See 3.10.0: Poisons and First Aid - DPX mounting medium
2.6 Peru balsam mounting medium
Purchase: Peru balsam oil, from Myroxylon pereirae, El Salvador
Peru balsam for microscopy, formerly from Myroxylon balsamum, Fabaceae, is used as a mounting medium for microscope specimens.

3.1 Acetic alcohol, biology fixative
Purchase: Acetic acid ACS reagent, 99.7% , CH3CO2H, (CH3COOH)
Purchase: Ethanol solution, 70% in H2O, Ethanolum 70 per centum, Ethyl alcohol, C2H5OH
Prepare this solution prepared immediately before use.
Prepare acetic alcohol solution: Mix 99 mL 70% ethanol with 99 mL concentrated ethanoic acid.

3.2 Aceto-orcein stain, microscopy stain
Purchase: Orcein, synthetic, Natural Red 28
Aceto-orcein stain, acetic orcein, ethanoic orcein, (stains chromosomes and nuclei crimson, cytoplasm pink), Harmful if ingested
Stains chromosomes and nuclei crimson, stains cytoplasm pink.
Prepare aceto-orcein stain:
1. Add l g synthetic orcein to 25 mL concentrated ethanoic acid, (glacial acetic acid), and 20 mL deionized water. Boil for 4 to 5 minutes in a narrow neck flask, fitted with a glass filter funnel to act as a condenser. Filter the solution while still hot. Add 5 mL concentrated ethanoic acid and stir to dissolve any orcein appearing on the surface of the mixture after filtration. Add 4 to drops of glycerol to retard evaporation.
2. Heat 50 cc 60% acetic acid until almost boils. Add 0.5 g orcein stain. Stir, cool and filter. Use freshly prepared solution.

3.3.0 Aceto-carmine, microscopy stain
Purchase: Schneider's aceto-carmine, Carmine solution
Prepare acetocarmine stain: Concentrated ethanoic acid, (glacial acetic acid), 45 mL, 0.5 g carmine, 55 mL deionized water.
3.3.1 Alizarin
Purchase: Alizarin, Dye content 97 %, 1,2-​Dihydroxyanthraquinone, Mordant Red 11
Alizarin, C14H8O4, 1,2-dihydroxyanthroquinone, Toxic, (aluminium ion indicator), Alizarin crimson, Mordant Red, Turkey Red, Rose madder, Alizarin red powder, (C.I. 58000), glucoside in Rubia tinctoria, Rubiaceae,
Sodium alizarin sulfonate, Used in assessing sweat function. Changes to purplish discoloration on contact with sweat.
Prepare alizarin solution:
Alizarin 2.4 g, (0.1 mM), Sodium carbonate anhydrous 2.4 g, (0.2 mM), Wheat starch BP, (pharmaceutical grade), 95.2 g.
Alizarin is anthraquinone derived and therefore carcinogenic. These chemicals are irritating to the eyes, respiratory system and skin. They definitely should not be inhaled or placed near the eyes. Prepare in a fume cupboard.

3.3.3 Alizarin Red S indicator
Purchase: Alizarin Red S, Alizarin Carmine, water soluble, Alizarin sodium monosulfonate, Alizarin sulfonate sodium, Alizarin sulfonic acid sodium salt, Sodium alizarin sulfonate, C14H7NaO7S
Alizarin Red S is an anthraquinone dye used to stain for calcium deposits, indicators of mature osteocytes. A 2% Alizarin Red S Stain forms a complex with calcium during the process of chelation, causing birefringence.
Indicator
pH colour change: 4.6-6.0, Acid: yellow, Base: red
Prepare Alizarin red S solution: 0.2 g in 100 mL of 1.5% HCl.

3.4 Aniline hydrochloride, microscopy stain
Purchase: Aniline hydrochloride, 97%, C6H5NH2
Aniline hydrochloride, aniline chloride, anilinium chloride,
Aniline hydrochloride is used to stain lignified tissue yellow
Prepare aniline hydrochloride stain: Make a saturated solution of aniline hydrochloride in deionized water. Filter then add a few drops of hydrochloric acid until solution is acid.
BE CAREFUL! Use gloves!

3.5 Aniline sulfate, microscopy stain
Purchase: Aniline sulfate, 98%, (C6H5NH2)2.H2SO4
Prepare aniline sulfate stain: Make a saturated solution of aniline sulfate in deionized water.
Filter then add a few drops of sulfuric acid until solution is acid.
BE CAREFUL! Use gloves! Highly toxic if ingested, Aniline sulfate, Solution < 3%, Not hazardous

3.5.1 Aniline blue stain
Purchase: Aniline Blue solution, 2.5% in 2% acetic acid, C32H25N3O9S3Na2
Aniline blue, (C.I. 42755), China blue, cotton blue, water blue, (alcohol soluble), Highly toxic if ingested
Aniline blue solution is used to stain collagen fibres blue in tissue sections using the Masson's trichrome protocol for staining multiple components.

3.5.2 Basic fuchsin microscopy stain
Purchase: Basic Fuchsin, Dye content >85 %, Basic Parafuchsin, Basic Red 9, Magenta™ O, Parafuchsin hydrochloride, Paramagenta hydrochloride, Pararosaniline chloride, Pararosaniline hydrochloride, Rosaniline hydrochloride, (C19H17N3. HCl)
Haematology and histology stain

3.5.3 Carbol fuchsin, microscopy stain
Purchase: Carbol Fuchsin,  Highly toxic if ingested
Carbol fuchsin, carbol-fuchsin, carbolfuchsin, stains mycobacteria, (acid-fast bacteria), in Ziehl-Neelsen stain, topical antiseptic, (Castellani's paint) .
Prepare carbol fuchsin: Dissolve 1 g of fuchsin in 10 mL of ethanol. Add this solution to 90 mL of 5% aqueous phenol then filter the solution.

3.5.4 Calberla's pollen stain
5 mL glycerol, 10 mL 95% ethanol, 15 mL distilled water, 2 drops of saturated aqueous solution of basic fuchsin
Stain pollen to light pink colour.

3.6 Carmine stain, microscopy stain
Purchase: Carmine powder, Alum lake of carminic acid, Cochineal, Nacarat, Natural Red 4
Carmine from cochineal scale insect.
Prepare carmine stain:
1. 4 g carmine, 1 mL concentrated hydrochloric acid, 15 mL deionized water. Boil gently in a fume cupboard for 10 minutes with continuous stirring. Cool then add 95 mL 85% ethanol, (alcohol). Filter before using the stain.
2. 1 g carmine + 2.5 g aluminium potassium sulfate in 500 mL distilled water. Boil for 20 minutes. Adjust final volume to 500 mL with distilled water. Filter and add a crystal of thymol as preservative. Refrigerate

3.8 DCIP, (2,6-Dichlorophenol-indophenol), microscopy stain
Purchase: DCIP, DPIP, 2,6-​Dichlorophenolindophenol sodium salt hydrate, redox indicator, C12H6Cl2NNaO2.xH2O
DCIP stain is used to show action of enzymes e.g. succinic dehydrogenase, chloroplasts when exposed to light and to test for vitamin C.
Prepare DCIP stain: Add 4 g carmine and 1 mL concentrated hydrochloric acid 15 mL deionized water. Boil gently in a fume cupboard for 10 minutes with continuous stirring. Cool, and add 95 mL 85% ethanol, (alcohol). Filter the solution. Add 1 g 2,6-Dichlorophenol-indophenol and 1 litre water.

3.10 Eosin, tetrabrornofluorescein, microscopy stain
Purchase: Eosin Y, Dye content ~99 %, Tetrabromofluorescein, Acid Red 87, Bromo acid J. TS, XL, or XX, Bromofluorescein, Bronze Bromo ES, Eosin yellowish, Solvent red 43, C20H8Br4O5
Counter stains are used to stain cell walls and cell contents. This is an animal tissue counter stain. Eosin stains cytoplasm. Eosin Y solution, aqueous or alcoholic or alcoholic, with phloxine is a general purpose cytoplasmic counterstain, used with haematoxylin and eosin staining.
Prepare eosin stain:
1. Use 1 g Eosin Y powder, 1000 mL 70% ethyl alcohol, (ethanol), 5 mL glacial acetic, (ethanoic), acid. Dilute 100 mL with 100 mL 70% alcohol, Add 2-3 drops of glacial acetic, (ethanoic) acid.
2. Dissolve 1 g eosin in 100 mL 70% ethanol.
3. Demonstrate fluorescence with 1:500 ethanol solution.

3.11.0 Gram stain, microscopy stain
Purchase: Gram Staining Kit for microscopy
Solutions required for the Gram stain procedure: 3.12 Gram's iodine solution
3.11.1 Crystal violet solution | 3.23 Safranin, microscopy stain.
Gram positive bacteria stain a blue purple colour because they retain the stain-iodine complex inside their cells.
Gram negative bacteria stain a red colour because they have cell walls that allow the stain-iodine complex to be washed out of the cell by alcohol.
Prepare Gram stain: Mix 2 g potassium iodide crystals and 1 g iodine crystals in 200 mL of deionized water, (Gram's iodine solution). Put the heat-fixed smear on the staining rack, cover with Crystal violet solution and leave for 1 minute. Tilt the staining rack and gently wash with water for 3 seconds. Flood the slide with Gram's iodine and leave for 1 minute. A stain iodine complex forms. Tilt and wash with water for a few seconds. Hold the slide at a 45 angle where the smear is clearly visible and run 95% alcohol, as a decolorizing agent, (Gram stain decolorizing solution), down the smear until no more colour runs out after 2-10 seconds. Wash with water. Counterstain by flooding the smear with safranin for 1 minute. Wash, blot dry and examine under an oil immersion lens. Gram stain is used routinely as a differential stain, so bacteria can be classified: "Gram positive" or "Gram negative".

Gram stain decolorizing solution
Prepare Gram stain decolorizing solution
1. Acetone / ethanol, (50:50 v / v), 0.1% basic fuchsin solution.
2. 50% ethanol, 50% acetone, basic fuchsin solution.
3. Dissolve 0.2 g of safranin or basic fuchsin in 10 mL 95% ethanol. Mix with 90 mL, of deionized water. The solution is stable and can be stored for months.

3.11.1 Crystal violet microscopy stain
Purchase: Crystal violet solution, C25H30ClN3
Crystal violet solution 1%, aqueous solution is used in Brown-Hopps method for Gram-positive and Gram-negative bacterial staining.
Crystal violet (C.I. 42555, C.I. basic violet 3), C25N3H30ClN3, "gentian violet", "methyl violet 10B", Toxic if ingested,
Crystal violet 10% W / V alcoholic
Use as 0.02% solution in water
Prepare crystal violet stain:
1. Dissolve 0.4 g crystal violet in 20 mL 95% ethanol. Mix with 80 mL 1% aqueous ammonium oxalate. Leave to stand for 48 hours before use. The solution is stable and can be stored for months.
2. For Solution A, dissolve 2 g crystal violet in 100 mL absolute alcohol. For solution B, dissolve 1 g ammonium oxalate in 100 mL deionized water. Add 25 mL Solution A to 100 mL Solution B.
3. For solution A, dissolve 1 g of crystal violet in 20 mL 95% ethanol. For solution B, add 0.8 g of ammonium oxalate to 80 mL deionized water. Add solution A to solution B and filter.
3.12 Ziehl–Neelsen stain, (acid-fast stain)
Purchase: Carbol-Fuchsin solution according to Ziehl-Neelsen, for microscopy
Bacteria stain bright red due to retention of carbol-fuchsin dye. Background is methylene blue counterstain.
It stains bright red mainly Mycobacteria, e.g. Mycobacterium tuberculosis that causes tuberculosis, (TB). It contains carbol-fuchsin, (aqueous solution of phenol + alcoholic solution of fuchsin), acid alcohol, (ethanol and hydrochloric acid), and methylene blue. The acid-fast mycobacteria are usually straight rod-shaped bacilli, 1 to 10 µm long by 0.2 to 0.6 µm wide. The high lipid content of their cell wall makes the bacteria resistant to penetration by many dyes and chemicals, including Gram stain. The procedure is to stain all the bacteria on the slide with hot carbol-fuchsin, then wash the slide with acid-alcohol. The remaining bacteria that are still red are the "acid-fast bacteria". Then counterstain with methylene blue dye, (Swiss blue), so that the acid-fast bacteria remain red and any other bacteria are stained blue.

3.13.0 Haematoxylin, C16H14O6, microscopy stain
Purchase:
Haematoxylin Solution, Gill No. 1 General purpose nuclear stain, progressive type, used with haematoxylin and eosin staining.
Haematoxylin Solution, Harris Modified General purpose nuclear stain, regressive type, used with haematoxylin and eosin staining.
Haematoxylin Solution, Mayer's General purpose nuclear stain, progressive type, used with haematoxylin and eosin staining.
Scott's Tap Water Substitute Concentrate is used as a "blueing reagent" in haematoxylin and eosin staining procedures.
Haematoxylin, haematoxylin, Natural Black 1, hydroxybrasilin, Toxic if ingested, Stains nuclei in plant and animal cells purple, blue or black. This stain was formerly "logwood" because it was made from the heartwood of the tree Haematoxylon campechianum that has a sweet taste and smells of violets. It is similar to natural dyes natural red 24 from brazilin, C16H14O5 and brazilein C16H12O5, from Caesalpinia echinata.
Prepare haematoxylin stain: Use a stock solution of 10% haematoxylin in 95% alcohol. The main alum haematoxylin solutions are Ehrlich's haematoxylin, Harris's haematoxylin, and Mayer's haematoxylin.
3.13.1 Haematoxylin solution, Delafield's haematoxylin, microscopy stain
Purchase: Haematoxylin, Natural Black 1, C16H14O6.xH2O
Prepare Delafield's haematoxylin stain:
1. Dissolve 4 g powder in 25 mL absolute ethanol. Mix gradually into 400 mL saturated aqueous alum, NH4Al(SO4)2.12H2O. Leave to stand for 3-5 days with a cotton plug in flask, exposed to direct light. Filter, then add 100 mL glycerine and 100 mL methanol. Leave to stand for at least 6 weeks.
2. Dissolve 1 g haematoxylin in 6 cc absolute alcohol. Add this drop by drop to 100 mL saturated ammonium alum. Leave in light and air for one week. Filter than add 25 mL glycerine and 25 mL methyl alcohol. Leave to stand until dark colour. Filter again.

3.13.2 Haematoxylin solution, Ehrlich's haematoxylin, microscopy stain
Purchase: Ehrlich's solution, (4-​(Dimethylamino)​benzaldehyde - hydrochloric acid solution)
Ehrlich's haematoxylin contains the oxidant sodium iodate.
Prepare Ehrlich's haematoxylin stain:
Mix 100 mL water, 100 mL ethanol, (absolute alcohol), 100 mL glycerol, 10 mL glacial acetic acid, 2 g haematoxylin, then add excess alum. aluminium potassium sulfate, AlK(SO4)2.12H2O to leave undissolved alum in the bottom. The solution is ready to use when it has a dark red colour.

3.13.3 Haematoxylin solution, Harris modified, microscopy stain
Purchase: Haematoxylin Solution, Harris Modified
Harris modified haematoxylin contains mercuric oxide.
Prepare haematoxylin solution, Harris modified, stain
Heat to dissolve. Add 50 ml of 10% alcoholic haematoxylin solution and heat to boil for 1 minute. Remove from heat and slowly add 2.5 g of mercuric oxide, (red). Heat to the solution and until it becomes dark purple colour. Cool the solution in cold water bath and add 20 ml of glacial acetic acid , (concentrated). Filter before use. General purpose nuclear stain, regressive type. Used with haematoxylin and eosin staining, (H & E stain), often called the "standard stain" or "routine stain" because most sections are routinely stained with it for a first inspection of the tissue being examined.
Purchase: Eosin-Hematoxylin solution according to Ehrlich for microscopy

3.13.4 Haematoxylin solution, ,Mayer's microscopy stain
Purchase: Haematoxylin Solution, Mayer's
Mayer's haematoxylin contains the oxidant sodium iodate.
Prepare Mayor's haematoxylin stain
1. Dissolve 50 g aluminium potassium sulfate, (alum), in 1000 ml distilled water. When alum is completely dissolved, add 1 gm haematoxylin. When haematoxylin is completely dissolved, add 0.2 gm sodium iodate and 20 ml acetic acid. Bring solution to boil and cool, and filter.
2. Dissolve alum in distilled water. When alum is completely dissolved, add haematoxylin. When haematoxylin is completely dissolved, add sodium iodate and acetic acid. Bring to boil and cool. Filter if it is necessary.
3.13.5 Haematoxylin solution, Heidenhain iron haematoxylin, microscopy stain
Heidenhain's iron haematoxylin is used to stain mitotic figures in amoeba, after fixation.
Prepare Heidenhain iron haematoxylin stain
1. Part 1. 4 g FeNH4(SO4)2.12H2O, (ferric alum, i.e. iron (III) ammonium sulfate) 100 mL deionized water,
Part 2. 10 g haematoxylin, 100 mL 95% ethanol. Mix equal quantities of Part 1. and Part 2.
The mixture is useful for a few hours only. Solution Part 1. is used as a mordant and Part 2. is for staining.
2. Incubate in 2% ammonium ferric sulfate (NH4Fe(SO4)2.12H2O). Then rinse in distilled water and then tap water. Then incubate in 0.5% haematoxylin in 2% ammonium ferric sulfate. Then wash in tap water and mount on microscope slide.

3.13.6 Haematoxylin solution, Weigert's haematoxylin, Weigert's iron haematoxylin, microscopy stain
Purchase: Weigert's iron haematoxylin solution
Weigert's haematoxylin solution is used for animal tissue for acidic stain procedures. It resists decolorization better than aluminium based haematoxylin formulations.
Prepare Weigert's iron haematoxylin stain
Part A: 2.5 g iron (III) chloride FeCl3.6H2O, 4.5 g iron (II) sulfate FeSO4.7H2O, 2 mL hydrochloric acid, 298 mL deionized water
Part B: 1 g haematoxylin, 100 mL 95% ethanol. Mix 1 part of B to 3 parts of A just before use.
This mixture can be used for up to 3 weeks.

3.15 Lugol's iodine solution, aqueous iodine solution, microscopy stain
Purchase: Lugol solution, according to Lugol, Iodine / Potassium iodide solution
Prepare Lugol's iodine stain
1. Dissolve 6 g potassium iodide in 1000 mL water. Add 4 g iodine crystals.
2. Dissolve 5 g of iodine crystals and 10 g of potassium iodide in deionized water and make up to 100 mL.
For bacterial staining, dilute to 1 / 5 with water.
3. Dissolve 1 g iodine crystals and 2 g potassium iodide in 300 mL deionized water.

3.16 Karo syrup mountant, microscopy stain
"Karo" is a brand of corn syrup, Karo Light Syrup contains light corn syrup, salt, vanilla, Karo Dark Syrup contains light corn syrup, salt, molasses. Previously, Karo Corn Syrup used to be high fructose corn syrup. The "Light" in "Karo Light Syrup" refers to the colour type, not low calorie syrup although a Karo lower calorie corn syrup is also sold.
Prepare Karo syrup mountant: 40 mL clear Karo syrup, 40 mL deionized water. Add 2 small crystals thymol, (or phenol), (about 0.2 g), as a preservative.

3.17 Lactophenol, microscopy stain
Purchase: Lactophenol blue solution, for microscopy, for staining moulds, Lactophenol Anilin Blue solution, Lactophenol Cotton Blue solution
Prepare lactophenol stain: Dissolve 25 g phenol in 50 mL water. Add 25 mL lactic acid. Add 50 mL glycerine. Store away from light. BE CAREFUL! Use gloves!

3.18 Lactophenol cotton blue, microscopy stain
Purchase: Cotton blue, Methyl Blue, Acid blue 93, Aniline blue water soluble, Poirriers blue, Water blue
Cotton blue, aniline blue, Helvetia blue, acid blue 93, C.I. 42780, C37H27N3Na2O9S3, histology and fungus stain
Prepare lactophenol cotton blue stain: 100 mL lactophenol, l g cotton blue. Dilute 5 mL to 100 mL with lactophenol before use.

3.19 Leishman's stain, Wright's stain
Purchase: Leishman's stain, used as histology stain, Eosin-polychrome methylene blue, a Romanowsky-type dye, used to identify leukocytes, malarial parasites, and trypanosomes.
Purchase: Wright Stain, Modified, Popular haematology stain used for differentially staining the cellular elements of blood. Wright stain, 0.3%, buffered at pH 6.8 in methanol. Contains stabilizers and surfactant.
Use Leishman's stain, Wright's stain, stain for white blood cells microscopy.
The ready made dark green crystalline powder, Wright's stain, contains the red acid dye eosin and the blue basic dye methylene blue. Variants of Wright's stain include buffered Wright's stain, Gram's stain, Wright-Giemsa stain, Jenner's stain.)
Prepare Leishman's stain
1. Dissolve 0.15 g of Leishman's stain in 10 mL absolute alcohol in a flask. Plug the flask with cotton wool and heat in a water bath over an electric heater for 15 minutes.
2. Add 0.115 g Leishman's stain to 100 mL of pure methanol in a flask. Plug the neck with cotton wool and warm in a water bath for 15 minutes with occasional shaking.
3.19a Methylene blue
Purchase: Methylene Blue hydrate, C16H18ClN3S.xH2O
Prepare methylene blue stain: Methylene blue, (0.1%), dilute 0.1 g methylene blue in 100 mL deionized water, dilute 1 in 10.
3.21 Orange IV
Purchase: Orange IV, [ 4-​(p-​Anilinophenylazo)​benzenesulfonic acid sodium salt], Acid Orange 5, Tropaeolin OO
Orange IV, orange G, tropaeolin OO, microscopy stain, (acid-base indicator, 1.3): 1.3
Prepare orange IV botanical tissues counter stain: Dissolve 0.5 g Orange IV powder in 100 mL 95% alcohol.
0.1% solution as acid-base indicator, pH < 1.4 red to pH > 2.6 orange.

3.22 Phloroglucinol, (10% solution), microscopy stain
Purchase: Phloroglucinol, 99.0%, (1,3,5-​Trihydroxybenzene ), C6H6O3
Acid phloroglucin is used to stain lignin in plant cells bright red.
Prepare phloroglucinol solution: Dissolve 1 g phloroglucin in 100 mL 50% alcohol.

3.23 Safranin, microscopy stain
Purchase: Safranin O, Dye content 85 % , Basic Red 2, Cotton Red, Gossypimine, Safranin T, Safranin Y or A, C20H19ClN4
C.I. 50240, C.I. Basic Red 2, Toxic if ingested, stain for lignin and cell walls
Prepare safranin stain:
1. Dissolve 0.2 g safranin in 10 mL 95% ethanol. Mix with 90 mL, of deionized water. The solution is stable and can be stored for months
2. Dissolve 1 g safranin in 100 mL 50% by volume alcohol / water mixture.
3. Dissolve 0.5 g safranin in 100 mL deionized water.
4. For a general contrast stain for lignin and cell walls, dissolve 0.02 g safranin in 10 mL 5% ethanol. Add 100 mL water to form 1% solution.

3.24 Schulze's solution, (chlor-zinc-iodine), microscopy stain
Dissolve 20 g zinc chloride in 9.5 mL warm water. Cool, add drop by drop 1.5 mL of the following solution until a persistent precipitate of iodine forms: 0.5 g iodine, 1 g potassium iodide, 20 mL deionized water.

3.25 Toluidine blue microscopy stain
Purchase: Toluidine Blue O, Basic Blue 17, Blutene chloride, Methylene Blue T50 or T extra, Tolonium chloride, C15H16ClN3S
Toluidine blue, microscopy stain, C.I. 52040, (plant cells microscopy stain), Toxic if ingested
The stain works best in fresh plant material. Lignified plant cell walls stain blue-green. Unlignified plant cell walls stain pink-purple.
Prepare toluidine blue stain: Dissolve 0.05 g toluidine blue stain in 100 mL of water.

3.27 Methyl cellulose solution, methocel, (low substitution)
Purchase: Methyl cellulose, viscosity 1,500 cP, 2 % in H2O (20°C) (lit.) [Different viscosity solutions are offered.]
Methyl cellulose, E461 (vegetable gum), (HEALTH intestinal problems)
Methyl cellulose, 2-methoxyethanol, methocel, (low substitution), ethers based on cellulose, viscid solutions
Methyl cellulose is not digestible, not toxic. It dissolves in hot water not hot water because precipitates out. It is used as thickener, emulsifier, virus culture, electrophoresis solution, "slime" in movies, slow movement protozoa and microorganisms by increasing viscosity. Prepared chemically from cellulose + sodium hydroxide + methyl cellulose. Degree of substitution (DS) up to 3 OH groups per glucose molecule (low substitution - to high substitution).
Prepare methyl cellulose solution: Dissolve methyl cellulose in water, to slow protozoa for study under the microscope.

3.28 Giemsa stain
Purchase: Giemsa Stain, Modified Solution according to Giemsa, Azure eosin methylene blue, Giemsa solution
Purchase Giemsa stain powder of Giemsa stain, (solution), contains 50% MeOH
Giemsa stain is a mixture of methylene blue, eosin, and azure. It is used to stain red blood cell stain and for diagnosis of malaria parasites
Prepare Giemsa blood stain: Dissolve 3.8 g of Giemsa powder in 250 mL of methanol. Add 250 mL of glycerine . Leave to stand for two months before using.
3.30 Alcian blue stain
Purchase: Alcian Blue 8GX, powder, Alcian Blue, Ingrain Blue, C56H68Cl4CuN16S4
Purchase: Alcian Blue solution, 1% in 3% acetic acid, pH 2.5
Alcian Blue 8GX is used as a heteroglycan stain for neutral, sulfated and phosphated mucopolysaccharides and glycosaminoglycans in tissues, e.g. cartilage, extra cellular matrices.
Prepare Alcian Blue solution: 1% in 3% acetic acid, pH 2.5
3.31 Nuclear fast red stain
Purchase: Nuclear Fast Red solution, Kernechtrot Solution, C14H8NNaO7S,
Nuclear Fast Red, (4-​Amino-​9,10-​dihydro-​1,3-​dihydroxy-​9,10-​dioxo-​2-​anthracenesulfonic acid sodium salt), Calcium Red,
Nuclear fast red stain is used as a red nuclear counterstain.
Prepare Nuclear fast red stain: Nuclear fast red 0.1% in 5% aluminium sulfate.

3.32 Oil red stain
Purchase: Oil Red O solution, 0.5% in isopropanol, Solvent Red 27, Sudan Red 5B, C26H24N4O
Oil Red O stain for microscopy, {1-​([4-​(Xylylazo)​xylyl]​azo)​-​2-​naphthol}
Oil Red O stain is used to stain fat in tissue.
Prepare Oil Red stain: supersaturated solution of Oil Red O in isopropanol
3.33 Papanicolaou stain
Purchase: Papanicolaou Stain, OG-6
Papanicolaou Stain, EA 50 Papanicolaou stain imparts a characteristic range of coloration to exfoliative cells of vaginal, cervical, prostatic and other body secretions, allowing critical examination of nuclei and cytoplasmic components.
Papanicolaou stain is used for routine diagnostic cytology to aid in the identification and classification of exfoliative cells.
3.34 Fluorescence staining of cells and tissues
Secondary antibodies can be conjugated with fluorescent dyes without compromising antibody affinity or specificity. The dyes are  photostable and not pH-dependent. Their reagent solutions come with 2.5% normal horse serum. Sections or cells stained with these reagents can be mounted in mounting media to display their immunofluorescence.
4.1 Aceto-alcohol, biology fixative
Purchase: Ethanol, 200 proof (absolute), for molecular biology, Ethyl alcohol CH3CH2OH
Aceto-alcohol fixative is used for animal material.
Prepare: Absolute ethanol, (alcohol), 30 mL, ethanoic, (acetic), acid, glacial 10 mL, mix immediately before use and discard after 1 hour,

4.2 Ethanol solution
Purchase: Ethanol, 200 proof (absolute), for molecular biology, Ethyl alcohol CH3CH2OH
A 50% solution of ethanol / water solution is used to preserve biological specimens.
4.3 FAA, biology fixative
Formalin–acetic acid–alcohol (FAA) and formalin–propionic acid–alcohol (FPA) are used as general purpose biological fixatives
Prepare FAA solution, (FAA = formalin, alcohol, acetic acid)
Formalin, (40% methanal) 5 mL, ethanol 70% 90 mL, acetic acid, glacial 5 mL
Formalin-acetic acid solution: formaldehyde (37-40%), 10 mL distilled water 90 mL, glacial acetic acid 5 mL
Formal ethanol fixative solution: 5 mL formaldehyde, 45 ml 95% ethanol, (reagent alcohol)

4.4 Formaldehyde, CH2O, formalin, biology fixative
Purchase: Formalin solution, neutral buffered, 10% histological tissue fixative Case containers are polypropylene
Formalin is used as a general purpose histological fixative.
Formalin is the commercial name for 40% formaldehyde solution. 1% formalin = 1 mL formalin then dilute to 100 mL with water. Formaldehyde vapour irritates eyes and delicate body tissues. Biological specimens preserved in formaldehyde should be kept in sealed containers.
Prepare formalin solutions:
Formalin Solution (10%, unbuffered): formaldehyde (37-40%) 10 mL, distilled water 90 mL
Formalin Solution (20%, unbuffered): formaldehyde (37-40%) 20 mL, distilled water 80 mL
Formalin Solution (10%, buffered neutral): formaldehyde (37-40%) 100 mL, distilled water 900 mL, NaH2PO4 4.0 g, Na2HPO4 (anhydrous) 6.5 g
Formalin Solution (20%, buffered neutral): formaldehyde (37-40%) 200 mL, distilled water 800 mL, NaH2PO4 4.0 g
Na2HPO4 (anhydrous) 6.5 g

4.5 Formal saline, para formaldehyde
Formal saline is more effective than 10% formalin because it is isotonic so less damage to erythrocytes. This mixture of formaldehyde in isotonic saline was formerly widely used for routine histopathology prior to the introduction of phosphate-buffered formalin.
1. 91 % formaldehyde, 37% solution in water, used as a biological fixative for marine animals
2. Formalin, (40% methanal) 100 mL, sodium chloride 10% solution 7 mL, deionized water 83 mL.
3. 40% formaldehyde 100 mL, sodium chloride: 9 g, distilled water: 900 mL, The fixation time is 12 to 24 hours.
4.6 Zenker's fluid, biology fixative
Prepare Zenker's fixative:
1. potassium dichromate 2.5 g, mercury II chloride 5.8 g, deionized water 95 mL.
2. mercuric chloride, potassium dichromate, sodium sulfate, glacial acetic acid, water
Zenker's fixative: is a rapid fixative used for for trichrome stains. The fixed tissue must be washed to remove potassium dichromate and treated with iodine solution to remove mercuric chloride.

4.8 Carnoy's fluid, biology fixative
Carnoy's fluid is an insect-fixing fluid and is used for fixation of DNA, RNA, Nissl granules and glycogen.
Prepare Carnoy's fluid:
1. 95% alcohol 75 mL, chloroform 30 mL. glacial acetic acid 10 mL, mix well
2. ethanol (absolute) 60 mL, chloroform 30 mL, glacial acetic acid 10 mL, mix well
3. 1 g of ferric chloride in 24 mL of absolute alcohol, chloroform 12 mL, glacial acetic acid 4 mL

4.9 KAA, insect-ixing fluid
For soft body insects, use 5 mL of KAA solution.
Prepare KAA solution
1. Kerosene 10 mL, 95% alcohol 100 mL, glacial acetic acid 20 mL.
2. Kerosene 8 mL, 95% alcohol 77 mL, glacial acetic acid 15 mL

4.10 Kahle's fluid, insect-fixing fluid
95% alcohol 100 mL, glacial acetic acid 7 mL, formalin 40 mL.

4.11 Oudeman's fluid, insect-fixing fluid
70% alcohol 88 mL, glycerine 4 mL, glacial acetic acid 8 mL.

4.12 Lacto-alcohol, insect-fixing fluid
Lactic acid 40 mL, 98% alcohol 37 mL, water 23 mL.

4.13 Pampl's fluid, insect-fixing fluid
Glacial acetic acid 4 mL, water 30 mL, 40% formaldehyde solution 6 mL, 95% alcohol 15 mL.

4.14 Sugaring mixture, insect-fixing fluid
500 g treacle, 1 kg brown sugar, 300 mL beer, 5 mL rum. Boil until uniform thickness occurs.

4.15 Barber's relaxing fluid, insect-fixing fluid
95% alcohol 50 mL, water 50 mL, ethyl acetate 20 mL, benzol 10 mL.

4.16 CRAF, biology fixative
Chromic acid, 1% 40 mL, formalin, (40% methanal), 10 mL, ethanoic, (acetic), acid, glacial 5 mL, deionized water 5 mL.
4.17 Bouin's solution, biology fixative
Purchase: Bouin's solution, Bouin's fluid
Bouin's solution, Bouin's fluid, is a fixative for preserving soft and delicate structures and is used as a mordant in trichrome procedures.
Prepare Boiun's solution: picric acid (saturated) 75 mL, formaldehyde (37-40%) 25 mL, glacial acetic acid 5 mL

4.18 Decalcifying Solution
Purchase Decalcifying Solution-Lite, aqueous solution, bp 208 °F, light yellow colour
Decalcifying Solution is used for the decalcification of routine, immunohistochemical and bone marrow core specimens.
4.19 Differentiation Solution
Differentiation Solution is an acidified alcohol solution for the differentiation of regressive haematoxylin stains.
4.20 Glutaraldehyde solution, microscopy fixative
Purchase, Glutaraldehyde solution, Grade I, 25% in H2O, electron microscopy fixative, Glutaric dialdehyde solution, pentane-1,5-dial , OHC(CH2)3CHO, aqueous solution
Glutaraldehyde, C7H8 O2, pentanedial, a colourless liquid, with a strong odour, is an antimicrobial, bactericide, fungicide and a virucide. It is used to sterilize hospital equipment, prevent bacterial growth in water supplies, and as a fixative for tissues. Breathing it causes headaches.

6.0 Direct microscopic mounts or squash preparations
Using sterile technique, remove a small portion of the colony with an inoculation needle and mount in a drop of Lactophenol Cotton Blue on a clean microscope slide. Cover with a coverslip, squash the preparation with the butt of the inoculation needle and then blot off the excess fluid.

6.1 Calcofluor White with 10% KOH, identify fungi
Use for the direct microscopic examination of skin scrapings, hairs, nails and other clinic specimens for fungal elements. This as a very sensitive method. A fluorescence microscope with the correct ultraviolet filters is required.
Solution A: Potassium hydroxide reagent.
Potassium hydroxide 10 g.
Glycerine 10 mL.
Deionized water 80 mL.
Solution B: Calcofluor white reagent
Calcofluor white 0.5 g.
Evans blue 0.02 g.
Deionized water 50 mL.
Mix one drop of each solution on the centre of a clean microscope slide. Place the specimen in the solution and cover with a coverslip.

6.2 Cellotape flag preparations, identify fungi
An excellent technique for the rapid mounting of sporulating fungi because more of the reproductive structures intact.
1. Using clear 2 cm wide cellotape and a wooden applicator stick, (orange stick), make a small cellotape flag, (2 × 2 cm).
2. Using sterile technique, gently press the sticky side of the flag onto the surface of the culture.
3. Remove and apply a drop of 95% alcohol to the flag, to act as a wetting agent and also dissolve the adhesive glue holding the flag to the applicator stick.
4. Place the flag onto a small drop of Lactophenol cotton blue on a clean glass slide, remove the applicator stick and discard. Add another drop of stain, cover with a coverslip, gently press and mop up any excess stain.

6.3 Cornmeal agar, identify fungi
Use for routine cultivation and identification of fungi.
Cornmeal agar, (Oxoid CM 0103), 8.5 g deionized water 500 mL.
1. Mix dry ingredients into 100 mL water, boil remaining water.
2. Add boiling water to mixture and bring to boil.
3. Dispense for slopes.
4. Autoclave for 10 minutes at 120oC, remove and slope.

6.4 Cornmeal glucose sucrose yeast extract agar, identify fungi
Use for zygomycete sporulation
Cornmeal agar, (Oxoid CM 0103), 17 g.
Dextrose, (Glucose), 2g.
Sucrose 3 g.
Yeast extract 1 g.
Deionized water 1000 mL.
1. Mix dry ingredients into 100 mL water, boil remaining water.
2. Add boiling water to mixture and bring to boil.
3. Dispense for slopes.
4. Autoclave for 10 minutes at 120oC, remove and slope.

6.5 Czapek Dox Agar, identify fungi
Use for routine cultivation of fungi, especially Aspergillus, Penicillium, and non-sporulating moulds.
Czapek Dox Agar, (Oxoid CM97), 45.4 g.
Deionized water 1000 mL.
1. Soak the ingredients in small amount of water.
2. Bring remaining water to boil, add to soaking ingredients and bring to the boil again, stirring continuously.
3. Dispense for slopes as required.
4. Autoclave at 121oC for 10 minutes, remove and slope or pour for plates as required.

6.6 Indian ink mounts
Nigrosin, Acid black 2, Nigrosin water soluble, may be substituted for India ink for histological work.
For the direct microscopic examination of cerebrospinal fluid, CSF, for Cryptococcus species, place a drop of Indian ink on the specimen. Mix well with a sterilized loop, and cover with a coverslip. The best brands to use are "Pelikan" or "Talons" Indian ink.

6.7 Lactophenol Cotton Blue, (LPCB), identify fungi
Use for the staining and microscopic identification of fungi
Cotton Blue, (Aniline Blue), 0.05 g.
Phenol Crystals, (C6H5O4), 20 g.
Glycerol 40 mL.
Lactic acid, (CH3CHOHCOOH), 20 mL.
deionized water 20 mL.
This stain is prepared over two days.
1. On the first day, dissolve the Cotton Blue in the deionized water. Leave overnight to eliminate insoluble dye.
2. On the second day, wearing gloves add the phenol crystals to the lactic acid in a glass beaker. Place on magnetic stirrer until the phenol is dissolved.
3. Add the glycerol.
4. Filter the Cotton Blue and deionized water solution into the phenol / glycerol / lactic acid solution. Mix and store at room temperature.

6.9 Potassium hydroxide with chlorazol black, identify fungi
Use for the direct microscopic examination of skin scrapings, hairs, nails and other clinic specimens for fungal elements.
Potassium hydroxide 10 g
Coral Azole E Black, (0.1%) 10 mL.
Glycerol 10 mL.
Deionized water 80 mL.
Using sterile technique, remove a small portion of the specimen with an inoculation needle and mount in a drop of KOH on a clean microscope slide. Cover with a coverslip, squash the preparation with the butt of the inoculation needle and then blot off the excess fluid.

6.10 Potato dextrose agar, identify fungi
Use for routine cultivation and identification of fungi.
Potato dextrose agar 39 g
Deionized water 1000 mL
1. Soak potato dextrose agar in small amount of the water in a stainless steel jug.
2. Boil remaining water, add to soaking ingredients, bring to the boil, stirring constantly.
3. Dispense for slopes as required.
4. Autoclave at 121oC for 15 minutes. Remove and slope or pour for plates as required.

6.11 Rice grain slopes, identify fungi
Use to induce sporulation and differentiation
Polished rice grains
Deionized water
1. Place 1 / 2 teaspoon rice grains into wide neck 20 mL lass vials.
2. Add 8 mL deionized water to each vial.
3. Lid, then slope on racks ensuring rice grains are evenly distributed.
4. Autoclave racks at 121oC for 15 minutes.

6.12 Orcinol-Bial's reagent
Purchase: Orcinol, 97%, 5-Methylresorcinol, CH3C6H3-1,3-(OH)2, (3,5-dihydroxytoluene)
Prepare Orcinol-Bial's reagent: Dissolve 0.2g orcinol in 100 mL concentrated hydrochloric acid, (Be careful! corrosive!).
6.13 Loeffler serum medium
Loeffler serum medium is used for identification of Corynebacterium species, e.g. Corynebacterium diphtheriae, diptheria, proteolytic activity, enzymatic hydrolysis of proteins, as a grey-white background to show microorganism pigmentation. It contains gelatin peptone, beef extract, sodium chloride, dextrose and bovine serum.
6.14 MacConkey agar
It is a primary plating medium, used to differentiate members of the coliform group. It is selective and differential. It select for gram negative bacteria because it contains bile salts and the dye crystal violet, which inhibit the growth of gram positive bacteria., It differentiates because it contains lactose, which gram negative bacteria can ferment lactose to form acid end products shown by a pink colour from the neutral red indicator. These pink staining colonies show the presence of coliform bacteria, an indicator of unsanitary food and water.

7.0 Prepare acids and bases
molarity 1 × volume 1 = molarity 2 × volume 2, M1V1 = M2V2
Dilute acids
Acetic acid 3 M: Dilute 172 mL of 17.4 M acid to 1 litre of water, (99 -800px acetic acid, ethanoic acid)
Hydrochloric acid 3 M: Dilute 258 mL of 11.6 M acid to 1 litre with water, (35% hydrochloric acid)
Hydrochloric acid 4 M: Dilute 400 mL of 10 M acid to 1 litre of water - for normal class use.
Nitric acid 4 M: Dilute 240 mL of 15 M acid. to 1 litre water - for normal class use.
Nitric acid 3 M: Dilute 195 mL of 15.4 M acid to 1 litre of water, (69% nitric acid)
Sulfuric acid 6 M: Dilute 168 mL of 17.8 M acid to 1 litre of water, (95% sulfuric acid)
Sulfuric acid 2 M: Dilute 112 mL of 35 M in 800 mL water, then add water to 1 litre - for normal class use.
Dilute bases
Ammonia solution 4 M: Dilute 220 mL, (28% ammonia), 18 M concentrated solution to 1 litre of water, ("ammonium hydroxide")
Ammonia solution 3 M: Dilute 200 mL, (28% ammonia), 14.8 M concentrated solution to 1 litre of water.
Ammonia solution 2 M: Dilute 330 mL, (10% ammonia), 6 M concentrated solution to 1 litre of water - for normal class use.
Potassium hydroxide 4 M: Dissolve 220 g KOH sticks in water, dilute to 1 litre of water - for normal class use
Sodium hydroxide 3 M. Dissolve 126 g the sticks, 95%, in water and dilute to 1 litre of water.
Sodium hydroxide 4 M Dissolve 160 g NaOH in 500 mL water, then dilute to 1 litre of water - for normal class use
Sodium hydroxide 8.5 M Dissolve 330 g NaOH in water, dilute to 1 litre of water, (For CO2 absorption)
Calcium hydroxide, (limewater)
1. 0.02 M. Saturated solution, 1.5 g Ca(OH)2 per litre, use some excess, filter off CaCO3, and protect from CO2 of the air.
2. Add 125 g of slaked lime, Ca(OH)2, to 3 litres of water, shake, allow precipitate to settle, siphon off clear liquid, and protect from CO2 of the air

8.0 Prepare salt solutions
Dissolve amount below then dilute to 1 L with water.
Aluminium chloride, AlCl3.6H2O, For 0.1 M solution, 24 g of hydrated salt in 1 L water
Aluminium sulfate, Al2(SO4)3.18H2O, For 0.l M solution, 66 g of hydrated salt in 1 L water
Ammonia, NH3 (aq) or NH4OH, For 2 M solution, dilute 330 mL of 10% solution in 1 L water
Ammonium chloride, NH4Cl, For 5 M solution, 270 g in water
Ammonium carbonate, (NH4)2CO3.3H2O, For 2 M solution, 300 g in 450 mL 10% NH3, then dilute in 1 L water
Ammonium iron (II) sulfate, For 0.1 M solution, 39.2 g in water, add 5 mL conc. H2SO4 in 1 L water
Ammonium oxalate, C2O4(NH4)2.2H2O, For 0.1 M solution, 16 g in 1 L water
Ammonium sulfate, (NH4)2SO4, For 0.1 M solution, 13.2 g in 1 L water
Barium chloride, BaCl2.2H2O, For 0.1 M solution, 24.4 g in 1 L water
Bismuth chloride, BICl3, For 0.17 M solution, 53 g in 1 litre of dilute HCl, 1 part conc. HCl to 5 parts water
Bismuth nitrate, Bi(NO3)3.5H2O, For 0.083 M solution, 40 g in 1 litre of dilute HNO3, 1 part conc. HNO3 to 5 parts water
Calcium chloride, CaCl2, anhydrous 0.l M solution, 11 g in 1 L water
Calcium chloride, CaCl2.2H2O, For 0.1 M solution, 14.7 g in 1 L water
Calcium hydroxide, Ca(OH)2, limewater, 10 g in 1 L water, shake, allow it to settle, decant clear liquid
Calcium nitrate, Ca(NO3)2, For 0.1 M solution, 16.4 g in 1 L water
Calcium sulfate, CaSO4.2H2O, For 0.1 M solution, Shake 10 g in 1 L water, leave to stand,
decant the clear liquid
Cobalt (II) chloride-6-water, CoCl2.6H2O, For 0.1 M solution, 23.8 g in 1 L water
Cobalt nitrate, Co(NO3)2.6H2O, For 0.1 M solution, 29 g in 1 L water
Copper (II) nitrate, Cu(NO3)2.6H2O, For 0.1 M solution, 29.6 g in 1 L water
Copper (II) sulfate, CuSO4.5H2O, For 0.1 M solution, 25 g in 1 L water + 5 mL conc. H2SO4
Iron (II) ammonium sulfate, Fe(NH4SO4)2.6H2O, For 0.5 M solution, 196 g in 1 L water + 10 mL conc. H2SO4, dilute to 1 litre
Iron (III) chloride, FeCl3.6H2O, For 0.1 M solution, 27 g in 1 L water + 20 mL HCl
Iron (III) nitrate, Fe(NO3)3.9H2O, For 0.1 M solution, 40.4 g in 1 L water
Iron (II) sulfate, FeSO4.7H2O, For 0.1 M solution, 27.8 g in 1 L water + 1 mL conc. H2SO4 to clear
Iron (III) sulfate, Fe2(SO4)3.9H2O, For 0.1 M solution, 56 g in 1 L water
Lead ethanoate, (CH3COO)2Pb.3H2O, For 0.1 M solution, 38 g in 1 L water + dilute ethanoic acid to clear
Lead nitrate, Pb(N03)2, For 0.1 M solution, 33 g in 1 L water
Magnesium chloride, MgCl2.6H2O, For 0.1 M solution, 20.3 g in 1 L water
Magnesium nitrate, Mg(N03)2.6H2O, For 0.1 M solution, 25.6 g in 1 L water
Magnesium sulfate, MgSO4.7H2O, For 0.1 M solution, 24.7 g in 1 L water
Manganese sulfate, MnSO4.H2O, For 0.1 M solution, 16.9 g in water
Nickel chloride, NiCl2.6H2O, For 0.1 M solution, 24 g in 1 L water
Potassium bromide, KBr, For 0.1 M solution, 12 g in 1 L water
Potassium carbonate, K2CO3, For 0.1 M solution, 13.8 g in water
Potassium chloride, KCl, For 0.1 M solution, 7.5 g in 1 L water
Potassium dichromate, For 0.1 M solution, 29.4 g in 1 L water, (K2Cr2O7)
Potassium dihydrogen orthophosphate, For 0.1 M solution, 13.6 g in 1 L water, (KH2PO4)
Potassium hydroxide, KOH, For 2 M solution, 110 g of KOH sticks in 1 L water
Potassium iodide, KI, For 0.1 M solution, 16.6 g in 1 L water
Potassium nitrate, KNO3, For 0.1 M solution, 10.l g in 1 L water
Potassium permanganate, KMnO4, For 0.1 M solution, 15.8 g in 1 L water
Potassium sulfate, K2SO4, For 0.1 M solution, 17.4 g in 1 L water
Silver nitrate, AgNO3, For 0.1 M solution, 17 g in 1 L water
Sodium borate, Na2B4O7.l0H2O, For 0.1 M solution, 38 g in 1 L water
Sodium carbonate, Na2CO3.10H2O, For 0.1 M solution, 28.6 g in 1 L water
Sodium carbonate, Na2CO3, (anhydrous), For 0.1 M solution, 10.6 g in 1 L water
Sodium chloride, NaCl, For 0.1 M solution, 5.8 g in 1 L water
Sodium chromate, Na2CrO4.4H2O, For 0.1 M solution, 23.4g in 1 L water
Sodium dichromate, Na2Cr2O7.2H2O, For 0.1 M solution, 29.8 g in 1 L water
Sodium ethanoate, CH3COONa.3H2O, For 00.1 M solution, 13.6 g in 1 L water, (sodium acetate)
Sodium hydrogen carbonate, NaHCO3, For 0.1 M solution, 8.4 g in 1 L water
Sodium iodide, NaI, For 0.1 M solution, 15 g in 1 L water
Sodium molybdate, Na2MoO4.2H2O, For 0.1 M solution, 24.2 g in 1 L water
Sodium nitrate, NaNO3, For 0.1 M solution, 8.5 g in 1 L water
Sodium nitrite, NaNO2, For 0.1 M solution, 7 g in 1 L water
Sodium oxalate, Na2C2O4, For 0.1 M solution, 13.4 g in 1 L water
Sodium sulfate, Na2SO4.10H2O, For 0.1 M solution, 32.2 g in 1 L water
Sodium sulfide, Na2S.9H2O, For 0.5 M 120 g in 1 L water
Sodium sulfite, Na2SO3.6H2O, For 0.1 M solution, 23.4 g in 1 L water
Sodium sulfite, Na2SO3, (anhydrous), For 0.1 M solution, 12.6 g in 1 L water
Sodium thiosulfate, Na2S2O3.5H2O, For 0.1 M solution, 24.8 g in 1 L water
Strontium (II) chloride, SrCl2.6H2O, For 0.1 M solution, 26.7 g in 1 L water
tri-Sodium phosphate, Na3PO4.12H2O, For 0.1 M solution, 38 g in 1 L water
Tin (II) chloride, SnCl2.2H2O, For 0.5 M solution, 113 g in 170 mL conc. HCl, dilute to 1 L + add tin foil
Tin (IV) chloride, SnCl2.5H2O, For 0.1 M solution, 35 g in 1 L water
Zinc sulfate, ZnSO4.7H2O, For 0.1 M solution, 28.8 g in 1 L water
9.1.1 Flour glue
Flour glues may be edible but they are not approved foods. They are safe for use by children but they should not be encouraged to consume them.
Combine 1 cup flour, 1.5 cups water, 1 / 3 cup sugar, 1 teaspoon vinegar. Remove any lumps and keep in a closed container.
9.1.2 Milk glue
Combine 1 cup water, 8 teaspoons powdered milk, 4 teaspoons vinegar, removes curds with a sieve, remove limps combine with 1 teaspoon baking soda solution.
9.1.3 Wallpaper paste
1 cup flour, 3 teaspoons alum, water to form appropriate consistence, 10 drops oil of cloves as preservative. Remove any lumps and keep in a closed container.
9.1.4 Turpentine copying fluid
Mix 4 parts of water with 1 part of turpentine. Add a piece of soap the size of the thunb nail. Shake the mixture until the soap is dissolved. The soap forms a turpentine / water emulsion. that keeps the turpentine and water mixture which have different densities from separating into separate layers. Copy a newspaper picture or text by moistening the newspaper, place a blank sheet of paper over it then rub with a smooth convex object. Ink from the newspaper dissolves in photocopy  liquid to make a picture in reverse.

9.2.0 Commercial glues and pastes, (in plastic containers)
Aerosol spray adhesive, sensitive spray glue, for binding textile, fabrics, paper, cardboard, flat PVC, foils, urethane / rubber foams, cork, pictures, photograph mounting, 330 g can
"Bostik" kids paste, non-toxic, with brush, 250 mL bottle
"Bostik" clear gum, non-toxic, bonds paper and cardboard, 5 litre
"Bostik" kids PVA, non-toxic, non-staining dries clear easily wash off clothes, 1 kg, 5 litre bottle
"Clag" paste, non-toxic, easy to apply with brush applicator, for cut and paste and papier mache, dries after 10-20 minutes, 150 g, 300 g, 5 kg bottle
"Clag", celmix, non-toxic adhesive powder, for finger painting, thickening PVA, hardboard sealer, 500 g container
"EC Mix-a-paste", non-toxic, multipurpose adhesive powder, 500 g
"EC Mix-it", instant papier mache, just add water, 2 kg pack
"EC" Tintex craft paste, non-toxic, non-staining, cellulose powder, adhesive for papier mache or gelling base for finger paint, 500 g tin
PVA glue, witches hat lid, kids school glue, non-staining, non-toxic, general classroom use, 250 mL bottle
PVA glue, Helmar, professional woodworking glue, 5 litre bottle
"UHU" craft glue, PVA, non -toxic. quick setting, washable adhesive, for hobby, arts and craft, general all purpose water resistant, 33 mL
"UHU", WOW glue stick, Goes on Purple, coloured overlay, dries clear, safe non-toxic, acid free, washable PVP glue stick, suit young children, 36 g
Accessories
Double -sided tape, high tack, acid and solvent free, 6 mm wide × 50 m roll
Glue brushes, flat, 15 mm wide, pack / 24
Paste spreaders, plastic paddles, 130 mm, pack / 24
Sellotape sticky dots, removable, clear double sided pack / 64, 1600

9.2.11 Prepare sterile media or solutions
Equipment: 1 autoclave or pressure cooker, 1 conical flask, 300 mL to 2.0 L according to quantity, culture tubes or test-tubes, sealed with cellulose bungs, Petri dishes, pipette and filler, 5 mL, sterile, pipette aid, 1 pH meter, 1 spatula, 1 set of scales, 1 piece of weighing paper, 1 pair of insulated gloves
Agar media:
1. Weigh out the individual components used for the manufacture of the culture medium onto a piece of weighing paper and place them into a conical flask. The quantities referred to below are for a 1 litre solution. If less culture medium is required, reduce the proportions of the individual components accordingly. The size of the conical flask depends on the amount to be prepared, 300 mL conical flasks are, for example, required for 200 mL culture medium.
2. Swirl the preparation around. until the soluble components have dissolved. Determine the pH and adjust it if necessary with 1 M NaOH or 1 M HCl, according to the values referred to below. 3. Seal the conical flask with a cellulose bung and autoclave in a pressure cooker at 121oC and 1 bar excess pressure for 20 minutes. Remove the conical flask from the pressure cooker using protective gloves and cool the flask to 50o.
3. This process can be accelerated by placing the flask under running water. The temperature has been reached when the bare back of the hand can touch the outside of the vessel without an unpleasant sensation.
4. Fill sterile plastic Petri dishes with the still liquid medium so that the base of the Petri dish is covered. Lift the lid of each Petri dish only for a short period of time so that no germs from the air get into the culture medium.
5. Place the filled Petri dishes in a safe place until the agar has set completely.
6. Inoculate the culture medium plates with material that contains bacteria, according to experimental needs.

9.2.12 Liquid broth media
1. Prepare the components required for the production of culture media according to the description for agar media, steps 1 through 3, autoclave, and cool down.
2. Fill culture tubes that have been sterilized in a drying cabinet, 180oC, 3 hours, with 5 mL of the culture medium, using sterile glass pipette and fillers. 3. Inoculate the culture tubes with material containing bacteria, as described in the experiments.

Media and solutions
9.2.14 Basal agar medium
Components: glucose 10.0 g / litre, casein peptone 4. 0 g / litre, meat extract 4.0 g / litre, yeast extract 0.5 g / litre, liver extract 0.5 g / litre, NaCl 2.5 g / litre, agar 11.0 g / litre, (pH 7.2).
9.2.15 Basal broth medium
The composition as for basal agar medium above, without agar. This is a liquid medium for overnight cultures.
9.2.16 Glucose nutrient agar
Purchase: Potato glucose agar
Prepare glucose nutrient agar:
1. Use ready made medium: glucose 1. 0 g / litre, nutrient agar 20.0 g / litre
2. Glucose 1.0 g / litre, peptone from meat 1. 0 g / litre, meat extract 3.0 g / litre, agar 12.0 g / litre, (pH 7.0).
9.2.17.0 Malt extract agar medium
Purchase Malt Extract Agar for microbiology
Use for malt extract agar medium for routine cultivation and identification of fungi.
Prepare malt extract agar medium:
1. Use ready made: malt extract agar 48.0 g / litre
2. Malt extract 30.0 g / litre, peptone from meat 3.0 g / litre, agar 11.0 g / litre, (pH 1.6), autoclave for 10 minutes
3. Dissolve 15 g malt extract and 18g bacteriological agar in 1 litre of deionized water. Dispense into bottles and sterilize with an autoclave.
4. Dissolve 20 g Oxoid Malt Extract malt extract 1000 mL of deionized water in a plastic beaker and pH the solution to pH 6.5 with NaOH. 2. Soak 20 g Bacto Agar in small quantity of solution. Bring remaining solution to the boil, stirring constantly. Add to soaking agar. Bring to boil, stirring constantly. 4. Dispense for slopes as required. Autoclave at 121oC for 10 minutes, remove and slope or pour for plates as required.

9.2.17.1 Malt extract broth medium
This is a liquid medium for overnight cultures.
Purchase: Malt Extract Agar for microbiology
Preparation: As for malt extract agar medium without the agar.

9.2.18 Minimal agar medium
Purchase: Davis Minimal Agar for microbiology, (Minimal Agar, Davis)
Prepare minimal agar medium: K2HPO4 3.5 g / litre, sodium citrate.x2 H2O 0.5 g / litre, MgSO4.7H2O 0.1 g / litre, (NH4)2SO4 1.0 g / litre, glucose 2.0 g / litre, D, L-histidine 0. 2 g / litre, D, L-arginine 0.2 g / litre, thiamine-HCl 0.05 g / litre, agar 11.0 g / litre, (pH 7.2).
9.2.19.0 Nutrient agar medium
Purchase: Nutrient Agar for microbiology
Prepare nutrient agar medium: peptone from meat 1. 0 g / litre, meat extract 3. 0 g / litre, agar 12.0 g / litre, (pH 7.0).

9.2.19.1 Starch nutrient agar medium
Prepare starch nutrient agar: Heat 4 g of soluble starch in 100 mL deionized water. Leave to cool then mix the suspension with 100 mL of hot liquid nutrient agar. Sterilize the mixture at 120oC for 15 minutes.

9.2.19.2 Milk agar medium
When a milk agar plate is made, it is assumed that the microbial population of the milk will not affect the experiment. However, the uninoculated area of the plate acts as a control.
Prepare sterilized nutrient agar, leave to cool to 45-50oC, add 10% pasteurized milk, skimmed, semi-skimmed or full cream milk.

9.2.20 Nutrient broth medium
Purchase: Nutrient Broth, No. 1, for microbiology , Standard - Nutrient Broth
Nutrient broth medium is a liquid medium for overnight cultures. It may also be purchased as a powder.
Prepare: broth 8.0 g / litre, or from individual components: The composition is the same as nutrient agar medium without the agar.

9.2.21 Urea agar medium
Purchase: Urea Test Agar, Christensen's Urea Agar, Urea Agar Base according to Christensen
Prepare using ready made medium:
1. urea agar, (Christensen), 21.0 g / litre, (pH 6.8)
2. urea 20.0 g / litre, deionized water 50 mL. Sterilize 2. with sterile filter, autoclave 1. and cool to 50oC, add 1.2 to 1.1) Components of urea agar: peptone from meat 1. 0 g / litre, glucose 1. 0 g / litre, NaCl 1. o g / litre, K2HPO4, 1.0 g / l, phenol red 12 mg / litre, agar 12.0 g / litre, (pH 6.8).
9.2.22 Vinegar bacteria medium
Vinegar bacteria medium is a liquid medium for overnight cultures.
Prepare vinegar medium: Peptone from meat 3. 0 g / litre, yeast extract 1. 0 g / litre, mannitol 21.0 g / litre.

9.2.23 MS agar medium
Purchase: MRS Agar for microbiology, Lactobacillus Agar
MS agar medium is used for Lactobacillus culture
Prepare MRS Agar:
1. With ready made medium: MS powder, (basal salts with minimal organics), 4.7 g / litre, granulated sugar 30.0 g / litre, agar 8. 0 g / litre, phytohormone solution, (e.g. BAP). Regulate pH with 1 M KOH.
2. Agar 12 g / litre, diammonium hydrogen citrate 2 g / litre, dipotassium hydrogen phosphate 2 g / litre, glucose 20 g / litre, magnesium sulfate 0.1 g / litre

9.2.24 BAP medium
Purchase BAP, 6-Benzylaminopurine, 6-BAP, BA, N6-Benzyladenine, C12H11N5, 100 mg, 1 M HCl 20 mg , plant cell culture-tested
BAP is a synthetic cytokinin that with auxins assists plant growth and development, so it is used in plant growth media, e.g. Murashige medium, Skoog medium, Gamborg's medium, Chu's N6 medium. Aso, BAP inhibits respiratory kinase in plants, and so increases post-harvest life of green vegetables.
Prepare BAP solution: Add 100 mg BAP to 100 mL of boiling deionized water, then add 1.0 mL of this stock solution to a culture medium, e.g. MS agar medium.
9.2.25 Buffer reagent, phosphate buffer reagent
Purchase: Phosphate Buffer pH 6.6 at 25 °C For use with Wright Stain
Purchase: Phosphate buffer pH 7.2 at 25 °C for haematology and histology staining techniques. For use as a buffer in Romanowsky type staining procedures, e.g. Wright Stain, Wright Giemsa, Giemsa, May Grunwald, Jenner and Leishman.
Individual components for 1 M solution
Prepare phosphate buffer reagent:
1. K2HPO4, 17.4 g / 100 mL,
2. KH2PO4, 12.9 g / 100 mL
Composition of 1 M buffer reagent: Make a solution of bipotassium hydrogen phosphate, (solution 1.) and potassium bihydrogen phosphate, (solution 2.) separately, each time using 100 mL deionized water. Place 60 mL of solution 1 into a 100 mL conical flask and, using a pipette and filler, adjust the pH to 7.5 with solution 2.
Composition of 0.1 M buffer reagent: Place 5 mL of solution 1. into a 100 mL conical flask with 45 mL deionized water. Place 5 mL of solution 2. into another 100 mL conical flask with 45 mL deionized water. Adjust the pH of dilute solution 1. to 7.5 by adding dilute solution 2 with a pipette and filler.
9.2.26 Domestos, NaClO
"Domestos" is composed of saturated sodium hypochlorite solution.
Prepare 20% Domestos Solution: "Domestos" household cleaning fluid 20 mL. Add deionized water to 100 mL.

9.2.27 Nitrogen-free mineral salts agar
Purchase: Nitrogen-free mineral salts agar medium
Nitrogen free mineral salts agar is used to culture a free-living nitrogen-fixing bacterium, (Azotobacter), from the soil.
Prepare nitrogen free mineral salts agar: Dissolve 0.50 g of FeCl3.6H2O in 500 mL of deionized water. Add 2 g K2HPO4 + 0.25 g of MgSO4.7H2O + 10 g glucose. Add 0.1 M NaOH until pH = 8.3. Add this solution to a mixture of 7.5 g agar and 1 g CaCO3. Autoclave the mixture at 121oC for 20 minutes then pour into Petri dishes to make a nitrogen free mineral salts agar plate.

9.2.28 Salt Solution
Components: K2HPO4, 3.8 g / litre, KH2PO4 1.2 g / litre, MgSO4.7H2O 1.1 g / litre, NaCl 2.5 g / litre, Fe2(SO4)3.4H2O 0.05 g / litre, Mn2(SO4)3 4H2O 0.05 g / litre, "Tween 80" 1 mL / litre, (pH 7.0).

9.2.29 Nutrient gelatin
Purchase: Nutrient Gelatin for microbiology
Nutrient Gelatin for microbiology is used for food control media, liquid media / broths, solid media / agars, biochemical identification media tests, gelatine liquefaction tests, identification tests & reagents, identification of proteolytic bacteria, soil / agriculture / environmental media, and media for cocoa, chocolate, sweets, food control, meat control, sterility testing, water control.

9.2.30 Mannitol yeast extract agar, (MYEA)
Purchase: Yeast mannitol agar for microbiology
Prepare mannitol yeast extract agar: Heat a mixture of 10 g agar in 1 litre of water until the agar is dissolved. Add 0.5 g K2HPO4, 0.2 g MgSO4.7H2O, 0.2 g NaCl, 0.2 g CaCl2.6H2O, 10 g mannitol and 0.4 g yeast extract. Autoclave the mixture at 121oC for 20 minutes then pour into Petri dishes to make MYEA plates.

9.2.31 Glucose nutrient agar medium
Purchase: Nutrient Agar
Prepare glucose nutrient agar: Add 0.5% (w / v) glucose to nutrient agar, dispense in bottles and sterilize.

9.2.32 DRBC, (food spoilage medium)
DRBC, Dichloran Rose-Bengal Chloramphenicol Agar is a selective medium for yeasts and moulds associated with food spoilage. It  inhibits bacterial growth and spreading moulds, e.g. Rhizopus and Mucor, but supports the growth of those species that cannot be otherwise isolated. The inhibition of spreading moulds and the general restriction of colony size results in improved counting and detection of mycotoxigenic moulds and other species of significance in food spoilage.

9.2.33 Nutrient agar, Microbiological base ingredients
Baird Parker Agar
Blood Agar (Base)
Brain Heart Infusion Agar
Brain Heart Infusion Broth
Brewer thioglycollate medium
Broth
Casein peptone Lecithin Polysorbate Broth
Casein hydrolysate broth
CASO Agar
Columbia Agar
Corn Meal Agar
HiCrome™ UTI Agar, modified
Mac Conkey Agar No 1
MacConkey Broth purple
Malt Extract Agar
Medium Base according to Loewenstein-Jensen
MRS Broth
MRS Agar
Mueller Hinton Agar
Mueller Hinton Broth
Mueller Hinton Broth 2 (Cation-Adjusted)
Nutrient Agar
Nutrient Broth No 1
Peptone Water, phosphate-buffered
Plate Count Agar
Plate Count Agar according to Buchbinder et al.
Potato Glucose Agar
R-2A Agar
Rappaport Vassiliadis Broth
Sabouraud 4% Glucose Agar
Sabouraud Glucose Agar with Chloramphenicol
Sabouraud Glucose Broth
Salmonella Chromogen Agar
SS-Agar
TBX Agar
TCBS Agar
Thiol broth
Todd Hewitt Broth
Tryptic Soy Broth
Tryptic Soy Agar
TSB Broth with Novobiocincin
Veal Infusion Broth
Vegitone Infusion Broth
Violet Red Bile Agar
Yeast Nitrogen Base