| REF No |
5026 |
| Purpose |
It is used to show Gram (+) and Gram (-) bacteria by light microscopy. |
| Ingredients |
A-Gram Crystal Violet Solution: Crystal violet, ethanol, ammonium oxalate, dimethyl sulphoxide.
B-Gram Lugol's Solution: Iodine, potassium iodide, 96% ethanol and PVP.
C-Gram Denatured Alcohol: Ethanol 96%, methanol, 2-propanol and stabiliser.
D-Gram Safranin Solution: Safranin O, methanol, triton X-100, and 96% ethanol, sodium azide. |
| Working Principle |
Gram Positive/Negative status is a feature depending on the cell wall (peptidoglycan) structure of bacteria. The peptidoglycan structure of Gram Negative bacteria is thinner between the outer membrane and the inner membrane than Gram Positive bacteria. There are a number of applications when performing this staining. These are primary staining, mordant application, selective application, contrast staining respectively. Primary staining is performed with crystal violet. Crystal violet is a basic dye used in staining methods. It is used for staining cellular parts in the technique. If this application is applied alone, all bacteria appear purple. Mordant application is to ensure that the dye enters the cell and binds to the peptidoglycan layer. Iodine solution is used as a colour fixing agent in this application. After this application, selective application method is applied. In this application, the preparation is treated with 96% alcohol and the outer membrane of Gram Negative bacteria is weakened and the dye is removed. At the end of this application, Gram Positive bacteria are purple and Gram Negative bacteria are colourless. The counter staining method is the last application. This application performed with safranin stains Gram Negative bacteria and makes them appear pink. At the end of this application, Gram Positive bacteria are still purple. |
| Warnings, Precautions, Recommendations |
Highly flammable liquid and vapour. Causes severe eye irritation. Suspected to cause cancer. Keep away from heat/ sparks/ flames/ hot surfaces. Wear protective gloves/ protective clothing/ eye protection/ face protection. In contact with skin: Immediately remove all contaminated clothing. Dry skin with water/ shower. In case of eye contact: Dry carefully with water for a few minutes. Remove contact lenses if fitted and easy to do so. Continue rinsing. In case of exposure or interaction IF: Seek medical advice/attention. Keep under lock and key. |
| Application
|
1. Cover the preparation with "A-Gram Crystal Violet Solution". After 5 minutes, shake off.
2. Delicately washed with distilled water.
3. Cover the preparation with "B-Gram Lugol's Solution". Pour off after 1 minute by shaking off.
4. Wash gently with distilled water.
5. Cover the preparation with "C-Gram Denatured Alcohol Solution" Remove the reagent by shaking the slide after 5-15 seconds.
6. Wash gently with distilled water.
7. Cover the preparation with "D- Gram Safranin Solution". After 1 minute, remove the reagent by shaking the slide.
8. Wash gently with distilled water.
9. Dry in air, preferably on a heated plate. Examine under a microscope. |
| Theoretical Results |
|
Gram positive bacteria |
Purple |
|
Gram negative bacteria |
Pink |
|
| Related Media
|
|
| Shelf Life |
2 years |
| Shelf Life After Opening the Lid |
|
| Storage Temperature |
15-25°C |
| pH (at 25°C) |
A-Gram Crystal Violet Solution: 5,0±0,3
B-Gram Lugol's solution: 5,0±0,3
D-Gram Safranin Solution: 6,8±0,2 |
| Transportation Information
|
Crystal Violet Solution
Carriage/shipment by road (ADR/RID) - UN1993, Ethanol, 3, II
Carriage/shipment by sea (IMDG) - UN1993, Ethanol, 3, II
Carriage/shipment by air (ICAO/IATA) - UN1993, Ethanol, 3, II
Gram Denatured Alcohol
Carriage/shipment by road (ADR/RID) - UN1987, (Ethanol, Methanol, Isopropyl alcohol) 3, II
Carriage/shipment by sea (IMDG) - UN1987, (Ethanol, Methanol, Isopropyl alcohol) 3, II
Air transport/shipment (ICAO/IATA) - UN1987, (Ethanol, Methanol, Isopropyl alcohol) 3, II
Gram Safranin Solution
Carriage/shipment by road (ADR/RID) - UN1992, (Ethanol, Methanol) 3, II
Carriage/shipment by sea (IMDG) - UN1992, (Ethanol, Methanol) 3, II
Air transport/shipment (ICAO/IATA) - UN1992, (Ethanol, Methanol) 3, II |
| Ordering and Packaging Information |
|
Order No |
Unit |
Volume |
Packaging Type |
Quantity in a Box |
Quantity in a Parcel |
|
445026050050 |
Piece |
50 mL |
50 ml, White Bottle with HDPE Dropper |
1 kit |
20 kits |
|
445026050250 |
Piece |
250 mL |
250 ml, PP White Bottle with Dropper |
1 kit |
15 kits |
|
445026020500 |
Piece |
500 mL |
500 ml, HDPE White Bottle 28 mm |
1 kit |
6 kits |
|
| Keywords
|
Gram-negative bacteria, Gram-positive bacteria, differentiation of bacteria, identification of bacterial isolates, staining technique in microbiology, Gram’s crystal violet Solution, Gram’s iodine Solution, Gram’s Decolorizer Solution, Gram’s safranin Solution, Gram staining, Gram's method |