Preparation of Gram Stain and Normal Saline
Preparation of Gram's staining reagents and 0.85% normal saline.
| Ingredient | Quantity |
|---|---|
| Crystal Violet | 20.0 g |
| Ammonium Oxalate | 9.0 g |
| Ethanol or Methanol | 95 ml |
| Distilled Water | To make 1 litre |
Preparation
In a clean piece of paper, 20 g of crystal violet is weighed and transferred to a clean brown bottle. Then, 95 ml of ethanol is added and mixed until the dye is completely dissolved. To the mixture, 9 g of ammonium oxalate dissolved in 200 ml of distilled water is added. Finally, the volume is made to 1 litre by adding distilled water.
| Ingredient | Quantity |
|---|---|
| Potassium Iodide | 20.0 g |
| Iodine | 10.0 g |
| Distilled Water | 1000 ml |
Preparation
To 250 ml of distilled water, 20 g of potassium iodide is dissolved. Then 10 g of iodine is mixed into it until it is dissolved completely. Finally, the volume is made to 1 litre by adding distilled water.
| Ingredient | Quantity |
|---|---|
| Acetone | 500 ml |
| Ethanol (Absolute) | 475 ml |
| Distilled Water | 25 ml |
Preparation
To 25 ml distilled water, 475 ml of absolute alcohol is added, mixed and transferred into a clean bottle. Then immediately, 500 ml acetone is added to the bottle and mixed well.
| Ingredient | Quantity |
|---|---|
| Safranin | 10.0 g |
| Distilled Water | 1000 ml |
Preparation
In a clean piece of paper, 10 g of safranin is weighed and transferred to a clean bottle. Then 1 litre distilled water is added to the bottle and mixed well until safranin is dissolved completely.
| Ingredient | Quantity |
|---|---|
| Sodium Chloride | 0.85 g |
| Distilled Water | 100 ml |
Preparation
The sodium chloride is weighed and transferred to a leak-proof bottle premarked to hold 100 ml. Distilled water is added to the 100 ml mark and mixed until the salt is fully dissolved. The bottle is labelled and stored at room temperature.
| Reagent | Main Components | Final Volume |
|---|---|---|
| Crystal Violet | Crystal violet + ammonium oxalate + ethanol/methanol | 1 litre |
| Lugol's Iodine | Potassium iodide + iodine | 1 litre |
| Acetone-Alcohol | Acetone + absolute ethanol + water | 1000 ml |
| Safranin | Safranin + distilled water | 1000 ml |
| Normal saline | Sodium chloride + distilled water | 100 ml |
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