| REF No |
5051 |
| Purpose |
Used for light microscopic examination of cytochemical staining of polymorphonucleocytes in bone marrow and blood. |
| Ingredients |
A) Glutaraldehyde Solution: 25% Glutaraldehyde, Buffers.
B) Acetone Acetone.
C) Tris Solution: Tris, Hydrochloric acid, Surfactant.
D) Diaminobenzidine Diaminobenzidine tetrahydrochloride
E) Hydrogen Peroxide Solution 3%: 35% Hydrogen peroxide, Acetanilide.
F) Copper Nitrate: Copper (II) nitrate.
G) Hematoxylin Solution Gill-3: Hematoxylin, Aluminum sulfate, Sodium iodate, Glacial acetic acid.
H) Gill's EA Solution: 95% Ethanol, Methanol, Fast green FCF, Eosin Y, Glacial acetic acid.
I) Scott's Concentrate Solution: Magnesium sulfate, Sodium bicarbonate. |
| Working Principle |
The Peroxidase (Myeloperoxidase) Leukocyte staining kit is based on the principle that leukocytes have granules containing the enzyme peroxidase. First, leukocyte cells on the preparation are reacted with a special reagent to enhance enzyme activity. This reagent activates the peroxidase enzyme and initiates a reaction by oxidizing the Hydrogen Peroxide present in the granules. As a result of this reaction, a colored compound is formed and the leukocyte cells are stained. |
| Warnings, Precautions, Recommendations |
Hazard Flammable liquid and vapor. May enhance fire; oxidizing. Long lasting, very toxic effect in aquatic environment. Take all precautions to prevent mixing with flammables/.... Causes severe eye irritation. May cause drowsiness or dizziness. Keep away from heat/sparks/flame/hot surfaces. - Do not smoke. Harmful if swallowed. Causes skin irritation. Causes allergic skin reactions. Causes serious eye damage. Inhalation may cause breathing difficulties, asthma attacks or allergies. Wear protective gloves/protective clothing/eye protection/face protection. IF INGESTED: If you feel unwell, call NATIONAL POISON ADVICE CENTER PHONE 114 or a doctor/physician. IN CASE OF CONTACT WITH SKIN: Wash with plenty of soap and water. IF INHALED: If breathing is difficult, remove the affected person to fresh air and hold in a comfortable position so that he/she can breathe easily. IN CASE OF CONTACT WITH EYES: Rinse carefully with water for several minutes. Remove contact lenses if worn and easy to do so. Continue rinsing. In case of skin irritation: Get medical advice/attention. Store in a well-ventilated area. Keep container tightly closed. Avoid release into the environment. |
| Application
|
1. To the container of DAB Tris Buffer Solution, add the 1% Hydrogen Peroxide Solution you prepared in the 0.5 ml tube. Label this as Oxidized DAB Tris Buffer Solution.
(CAUTION! Do not immerse the slide in this first).
2.Immerse the preparation in the Glutaraldehyde Fixative solution in your cooler. After 1 minute, shake it out.
3.Wash carefully with distilled water for 30 seconds.
4.Immerse in oxidized DAB Tris Buffer Solution. After 45 seconds, shake off.
5.Wash carefully with distilled water for 30 seconds.
6.Immerse the preparation in a container of Copper Nitrate Solution. Dip in and out of the container from time to time. After 2 minutes, shake it out of the container.
7.Wash carefully with running distilled water for 30 seconds.
8.Immerse the preparation in the G- Hematoxylin Solution Gill-3 container. Dip in and out of the container from time to time. After about 8-10 seconds, shake it out of the container.
9.Wash carefully with distilled water for 5 seconds.
10.Immerse the preparation in the Scott's Solution container. Dip in and out of the container several times. After 12 seconds, shake it out of the container.
11.Wash carefully with running distilled water for 5 seconds.
12.Immerse the preparation in the container of H-Gill's EA solution. After 1 minute, shake it out.
13.Immerse or wash twice for three seconds each in a 95% ethanol container.
14.Immerse or wash twice for three seconds each in a container of absoluethanol.
15.Immerse or wash three times for three seconds each in a container of xylene.
16.Cover with synthetic resin or others and examine under a microscope. |
| Theoretical Results |
|
Neutrophil peroxidase |
Gray to black intracellular granulation |
|
Lymphocytes |
No peroxidase activity |
|
Monocyte peroxidase |
Weaker gray to black intracellular granulation |
|
Eosinophils |
From orange to red |
|
Basophils |
Blue |
|
| Related Media
|
|
| Shelf Life |
1 year |
| Shelf Life After Opening the Lid |
|
| Storage Temperature |
2-8 °C |
| pH (at 25°C) |
A) Glutaraldehyde Solution pH: 7,6
|
| Transportation Information
|
|
| Ordering and Packaging Information |
|
Order No |
Unit |
Volume |
Packaging Type |
Quantity in a Box |
|
445051050050 |
Piece |
50 mL |
50 ml, White Bottle with HDPE Dropper |
1 kit |
|
| Keywords
|
Peroxidase (Myeloperoxidase) Leukocyte |