Introduction to the preparation of common dyeing liquids
Experimental procedure 1. Loeffler alkaline blue dye solution A: methylene blue 0.6g Lab Nitrile Gloves,Disposable Gloves,510K Disposable Gloves,Astm D6319 Examination Gloves Puyang Linshi Medical Supplies Co., Ltd. , https://www.linshihealths.com
95% alcohol 30ml
Solution B: KOH 0.01g
Distilled water 100ml
Prepare solution A and solution B separately, mix well and mix.
2. Ziehl carbolic acid red dyeing solution A: basic red complex (basic fuchsin) 0.3g
95% alcohol 10ml
Solution B: Carbolic acid 5.0g
Distilled water 95ml
After the alkaline red complex was ground in a mortar, 95% alcohol was gradually added, and the grinding was continued to dissolve to prepare a liquid A.
The carbolic acid is dissolved in water to form a liquid B.
Mix liquid A and liquid B to form. Usually, the mixture can be diluted 5-10 times, and the diluent is easily deteriorated, and it is not suitable for one time.
Third, Gram staining solution 1. Ammonium oxalate crystal violet dye solution A: crystal violet 2g
95% alcohol 20ml
Solution B: ammonium oxalate 0.8g
Distilled water 80ml
The two solutions of A and B were mixed and used after standing for 48 hours.
2. Lugol iodine iodine tablets 1.0g
Potassium iodide 2.0g
Distilled water 300ml
First dissolve potassium iodide in a small amount of water, then dissolve the iodine tablets in the potassium iodide solution. After the iodine is completely dissolved, add enough water to make it.
3.95% alcohol solution.
4. Safranine O (2.5 g)
95% alcohol 100ml
Take 10ml of the above prepared blush alcohol solution and mix it with 80ml of distilled water.
Fourth, spore staining solution 1. Malachite Green Malachite Green 5g
Distilled water 100ml
2. Safranin aqueous solution reddish 0.5g
Distilled water 100ml
3. Phenol fuchsin solution, basic fuchsin, 11g
Anhydrous alcohol 100ml
10 ml of the above solution was mixed with 100 ml of a 5% phenol solution, and filtered for use.
4. Melanin solution (water soluble melanin 10g)
Distilled water 100ml
Weigh 10g of melanin dissolved in 100ml of distilled water, put it in boiling water bath for 30 minutes, filter the filter twice, add water to 100ml, add 0.5ml of formaldehyde, and set aside.
5. Capsule staining solution 1. Melanin aqueous solution melanin 5g
Distilled water 100ml
Formalin (40% formaldehyde) 0.5ml
The melanin was boiled in distilled water for 5 minutes and then formalin was added as a preservative.
2. The saffron dye solution is the same as the saffron complex dye solution in the Gram stain.
Sixth, flagella dyeing solution A: tannic acid 5g
FeCl3 1.5g
Distilled water 100ml
Formalin (15%) 2.0ml
NaOH (1%) 1.0ml
After the match, the next day will be used poorly, and the third day will not be used.
Solution B: AgNO3 2g
Distilled water 100ml
After the AgNO3 was dissolved, 10 ml of the solution was taken out, and concentrated NH4ON was added dropwise to the remaining 90 ml of AgNO3 to make it a very thick suspension, and NH4OH was further added dropwise until the newly formed precipitate was dissolved again. Then, the spare 10 ml of AgNO3 was slowly dropped, and mist appeared, but after shaking gently, the misty precipitate disappeared again, and then AgNO3 was dropped until a slight and stable misty precipitate appeared after shaking. If the fog is not heavy, the dye can be used for one week. If the fog is heavy, the silver salt precipitates and should not be used.
Seven, Fulgen's nuclear staining solution 1. Schiff reagent Add 1 g of basic red complexion to 200 ml of boiling distilled water, shake for 5 minutes, cool to about 50 ° C for filtration, add 1 mol / L of HC 120 ml, shake. When cooled to 25 ° C, add 3 g of Na2S2O5 (sodium metabisulfite), shake it, place it in a brown bottle, wrap it in black paper, and place it in a dark place overnight. At this time, the reagent should be light yellow (if it is pink, it cannot be used). ), adding neutral activated carbon filtration, the filtrate is shaken for 1 minute, and then filtered, and the filtrate is placed in a cool dark place (note: the filtration needs to be carried out in the dark).
All utensils used throughout the operation need to be very clean and dry to eliminate reducing substances.
2. Schandium fixative solution A: a saturated aqueous solution of mercury (50 ml of mercury in water plus 95% alcohol 25 ml mixed).
Solution B: glacial acetic acid Take 1 ml of solution A, 9 ml of solution B, mix and heat to 60 °C.
3. 10% of the aqueous sulfite solution was added to 5 ml of an aqueous solution of sodium sulfite, 5 ml of 1 mol/L HCl, and 100 ml of distilled water was added to obtain a mixture.
Eight, Bouin's fixed solution of picric acid saturated aqueous solution 75ml
Formalin (40% formaldehyde) 25ml
Glacial acetic acid 5ml
1 g of picric acid can be made into a 75 ml saturated aqueous solution.
First dissolve the picric acid into an aqueous solution, then add formalin and glacial acetic acid to shake and serve.
Nine, lactic acid charcoal cotton blue dyeing solution carbolic acid 10g
Lactic acid (specific gravity 1.21) 10ml
Glycerin 20ml
Distilled water 10ml
Cotton blue (cottonblue) 0.02g
The carbolic acid is heated and dissolved in distilled water, then lactic acid and glycerin are added, and finally cotton blue is added to dissolve it.
X. Wright dyeing liquid Wright's dye powder 0.3g
Glycerin 3ml
Methanol 97ml
The dye powder is ground in a dry mortar, first added with glycerin, then added with methanol, placed in a glass bottle overnight, and filtered.
Eleven (Levowitz-Weber) dye solution (Experiment 62)
In a conical flask of 52 ml of 95% alcohol and 44 ml of tetrachloroethane, 0.6 g of met hylene blue chloride was slowly added, and the Erlenmeyer flask was shaken to dissolve. Put at 5-10 ° C for 12-24 hours, then add 4 ml of glacial acetic acid. Use good quality filter paper such as WhatmanNo. 42 or filter paper of the same quality. Store in a clean, closed container.