JorVet J-323 Gram Stain KitThis page contains information on JorVet J-323 Gram Stain Kit for veterinary use.
The information provided typically includes the following:
- JorVet J-323 Gram Stain Kit Indications
- Warnings and cautions for JorVet J-323 Gram Stain Kit
- Direction and dosage information for JorVet J-323 Gram Stain Kit
JorVet J-323 Gram Stain KitThis treatment applies to the following species:
Crystal Violet: Crystal Violet 0.5% w/v in Denatured Alcohol.
Iodine Solution: PVP Iodine 1.9% w/v; Potassium Iodide 13% w/v.
Decolorizer: Specially Denatured Alcohol/Acetone
Safranin Counterstain: Safranin 0.5% in denatured alcohol w/v.
JorVet J-323 Gram Stain Kit Indications
To differentiate gram positive and gram negative bacteria. For in vitro diagnostic use.
Sample Collection: Organisms being stained by the Gram method are usually taken from a solid or liquid medium on (in) which they have been cultured from their original source (e.g. wounds, throat swabs, sputum, etc.). An aqueous suspension is made, in the case of the solid medium, by taking a small amount of the material and suspending it in a drop of distilled water on a microscope slide. Care should be taken not to make the smear too thick. In the case of a liquid medium, a drop is used directly from the culture container. However, due to the solids from the medium, this method is not always satisfactory.
Directions For Use
The suspension made by either method is air dried then “fixed” by passing rapidly through a Bunsen burner flame two or three times. Allow the smear to cool before staining. (See Sources of Error section.)
Step by Step Procedure:
1. Allow smears to air dry thoroughly (until moisture is no longer visible) then “fix” by passing rapidly through a Bunsen burner flame two or three times. (See Sources of Error section.)
2. Place the “fixed” smears on a staining rack and cover completely with Crystal Violet (Reagent #1) for 30-60 seconds.
3. Wash off the stain with distilled water.
4. Cover the slide with Iodine Solution (Reagent #2) for 30 seconds.
5. Wash off with distilled water.
6. Decolorize (Reagent #3 for 10-15 seconds).
7. Wash thoroughly with distilled water.
8. Replace the slide on the stain rack and cover completely with Safranin Counterstain (Reagent #4) for 30-60 seconds.
9. Wash with distilled water, air dry and examine under immersion oil. Label clearly. Gram positive organisms stain a dark purple; gram negative organisms stain red.
Gram positive organisms such as cocci appear dark blue or black, and gram negative organisms, such as the coliforms and pseudomonads, appear pink. The nuclei of leukocytes stain pink. All fungi are gram positive.
Stability of Final Reaction: Stained smears, after being properly mounted with mounting medium, have been observed to retain their staining characteristics for at least two months.
Sources of Error:
1. Overheating (burning) during fixation can be avoided by just touching the back of the slide to the back of the hand each time the smear has been passed through the flame.
2. Do not stain smears which have only been air dried. Smears must also be “fixed”.
3. Smears should not be too thick. After air drying, examine under a microscope. If there are no areas of bacteria separation, more water should be added to dilute the smear. Repeat air drying and “fixing” as in step #1 of the Step by Step Procedure section.
4. After staining, it is essential that the back surface of slide is wiped clean.
5. If washing with distilled water is not done adequately, crystallization of stain may appear on the slide.
6. The manufacturer recommends that a known Gram positive and Gram negative control be stained at the same time as the test culture as an additional measure of quality control.
7. Staining times may vary to suit the individual.
8. In steps #3, #5 and #7 of the Step by Step Procedure section slides should be removed from the staining rack and held at an angle while washing.
#1. Crystal Violet: Store at room temperature.
#2. Iodine Solution: Store at room temperature.
#3. Decolorizer: Stable indefinitely. Store at room temperature.
#4. Safranin Counterstain: Store at room temperature.
#1. Crystal Violet: Poison: Do not take internally. Avoid contact with eyes. Vapor harmful. Flammable.
#2. Iodine Solution: Poison: Do not take internally. Avoid contact with eyes. Avoid breathing vapor.
#3. Decolorizer: Poison: Do not take internally. Avoid contact with eyes. Avoid breathing vapor.
#4. Safranin Counterstain: No special handling precautions.
Presentation: 4 x 250 mL bottles.
NAC No.: 11520030
1450 NORTH VAN BUREN AVE., LOVELAND, CO, 80538
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Copyright © 2014 North American Compendiums. Updated: 2014-05-28