Acid Fast Stain (Auramine O)

Assay-kits

Acid Fast stain (Auramine O)

Product code: ACMBS-05
Specifications: 4x250ml, 4x500ml (Box)
Intended use: Intended for fluorescence staining of Mycobacteria and other bacteria
Store temperature: 5℃～30℃
Expiry date: 24 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

When the acid-fast bacteria are stained by fluorescent dye Auramine O, it would appear bright orange or yellow-green color by the fluorescence microscope with purple and blue light activated filter plate (Wavelength: 350~490nm). It can work with low magnification, so it will find acid resistant bacteria quickly.

Main components

Reagent composition	Main ingredient
1. Auramine O solution	Auramine O
2. Acid Alcohol solution	Hydrochloric acid, Ethanol
3. Counterstain solution	Potassium permanganate

Sample requirement

Pick the sputum specimens of cheese-like, pus-like or suspicious part of about 0.05ml, evenly coated on the front of the slide into a 10mmx20mm oval sputum, natural dry. To stain after heating fixed.

Test procedure

1. After the smear is fixed, add Auramine O solution and stain for 15 minutes and wash it with water.
2. Decolorizing with acid alcohol solution for 1～2 minutes and wash, if it's necessary, then bleaching once, until it's colorless by naked eye to observe and wash.
3. Add counterstain solution dyeing for 2～4 minutes (duplicate dyeing time cannot be too long to avoid weakening the fluorescence brightness), wash.
4.Microscopic examination in a dark room after drying: First, a 10×eyepiece and a 20×objective lens is used for microscopic examination to find fluorescent rod-like substances suspected to be acid-fast bacillus and then use the 40×objective to confirm.

Result interpretation

The acid-resistant bacteria showed a bright orange-yellow or yellowish-green bar with a slightly curved, while the background was black and some of the background residues were pale yellow.

Method limitation

Only use for staining of acid-fast bacteria.

Precautions

1. All positive smears should be confirmed again by Kinyoun or Ziehl-Neelsen staining, the same smear with auramine O staining can also be used, but the result may be less ideal.
2. The Auramine O solution in this reagent is very stable and there is no need to worry about the problem of fluorescence reduction during the validity period.
3. The specimens should be observed immediately after being stained or stored in the shaded and cool environment within 24 hours, stored in the shaded and cool environment within 7 days can still be observed, but the fluorescence of positive bacteria will be weakened.
4. When storing reagents, try to avoid high and low temperature environments and direct sunlight.
5. This product should be used by professionals and interpretation of the results.
6. Before use, should read the instructions for use, use within the validity period, and do a good job of personal hygiene protection.
7. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
8. The production date, production batch number and expiration date are in the packaging.

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