Acid Fast Stain (Auramine O)

Acid Fast Stain (Auramine O)
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 compositionMain ingredient
1. Auramine O solutionAuramine O
2. Acid Alcohol solutionHydrochloric acid, Ethanol
3. Counterstain solutionPotassium 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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