Acid fast stain lab report. Complete Acid Fast Stain Lab 2022-10-13
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An acid fast stain, also known as a Ziehl-Neelsen stain or a carbolfuchsin stain, is a laboratory technique used to identify acid fast bacilli, a group of bacteria that includes Mycobacterium tuberculosis, the causative agent of tuberculosis. The acid fast stain is a differential stain, meaning it is used to differentiate between different types of bacteria based on their physical properties.
The acid fast stain is a two-step process that involves staining the bacteria with a primary dye and then decolorizing the stain with an acid-alcohol solution. The primary dye, carbolfuchsin, is a red or pink dye that is resistant to decolorization. When the bacteria are exposed to the primary dye, the dye is absorbed by the bacteria and the bacteria appear red or pink when viewed under a microscope.
The second step of the acid fast stain involves decolorizing the primary dye with an acid-alcohol solution. This step is necessary because some bacteria, such as non-acid fast bacteria, also absorb the primary dye. The acid-alcohol solution removes the primary dye from the non-acid fast bacteria, leaving only the acid fast bacteria stained red or pink.
To perform an acid fast stain, a sample of bacteria is first collected and placed on a microscope slide. The sample is then heat-fixed to the slide, which helps to preserve the bacteria and make them easier to stain. The primary dye is applied to the slide and allowed to absorb into the bacteria for a period of time. The slide is then rinsed with water and the acid-alcohol solution is applied to decolorize the stain.
After the acid fast stain is completed, the slide is examined under a microscope to identify the presence of acid fast bacilli. The bacteria appear red or pink when viewed under the microscope, while non-acid fast bacteria appear unstained.
The acid fast stain is a valuable tool for identifying acid fast bacilli, particularly Mycobacterium tuberculosis, which is a major public health concern. The acid fast stain is widely used in clinical laboratories to diagnose tuberculosis and other diseases caused by acid fast bacilli.
In conclusion, the acid fast stain is a laboratory technique used to identify acid fast bacilli, a group of bacteria that includes Mycobacterium tuberculosis. The acid fast stain is a two-step process that involves staining the bacteria with a primary dye and then decolorizing the stain with an acid-alcohol solution. The acid fast stain is a valuable tool for identifying acid fast bacilli and is widely used in clinical laboratories to diagnose diseases caused by these bacteria.
Acid fast stain lab report Free Essays
The primary stain used in acid-fast staining, carbolfuchsin, is lipid-soluble and contains phenol, which helps the stain penetrate the cell wall, this is further assisted by the addition of heat. The smear is then treated with a strong decolourizer, which strips the stain from all non-acid-fast organisms but does not permeate the cell wall of acid-fast organisms. Heat the slide to steaming for 5 minutes. Be sure to include the organism or specimen name and the magnification at which you viewed the slide. . PRE-ASSESSMENT PROCEDURE Prepare a mixed culture smear for Kinyoun Acid-Fast Stain For this Exercise: Use TSB broth of E.
Then gently rinse slide with distilled water. Because of this characteristic, carbol fuchsin stain is extremely helpful in identifying diseases caused by acid-fast bacteria, particularly Tuberculosis mycobacterium tuberculosis , Atypical Tuberculosis Mycobacterium avium, Mycobacterium bovis , Listeriosis Listeria monocytogenes , which is partially acid-fast , and Leprosy Mycobacterium leprae. The acid-fast stain is therefore a differential stain since it uses two dyes to distinguish two groups of organisms based on the mycolic acid in their cell walls. Sterilize the loop and let it cool. Why did his physician assume a different cause? Then remove paper and gently rinse slide. A prescribed number of microscopic fields are examined and the number of AFB is determined and reported using a standard scoring system.
Ferrous iron can be absorbed in the body system. Acid rain is rain that has been made acidic by certain pollutants in the air. In the United States, around 500-1,000 new cases of Nocardia infection occur annually. What can you conclude? Decolorize with acid-alcohol until the runoff is clear. There were modifications made by the instructor to dilute the 6M NaOH to 0.
4.1: Introduction to Acid fast and Endospore Staining
Wearing gloves and chemical eye protection, and the slide on the steaming apparatus, cover the smear s with a strip of bibulous paper cut slightly smaller than the slide and apply ZN carbolfuchsin stain. When dry, observe under oil immersion. Then gently rinse the slide with distilled water into the staining tray. Because you know ahead of time which organisms should give a positive result and which should give a negative result, it is okay to mix them into a single emulsion. The nonacid-cells are then counterstained with brilliant green to show the difference. Globally, there are approximately 10 million TB infections per year.
SARAH GILDENBERG LONE STAR COLLEGE - MONTGOMERY BIOL 2420 - SPRING 2020 Microbiology Lab Notebook Report Acid Fast Stain - Exercise 3-6 March 17, 2020 Introduction The acid-fast stain is a differential stain used to detect cells capable of retaining a primary stain when treated with an acid alcohol. It is an important differential stain used to identify bacteria in the genus Mycobacterium, some of which are pathogens e. Air dry and then heat-fix. The appearance of Non-Acid fast bacteria and other Cells in the Specimen — Green Colored if you are using the malachite green as counterstain or Blue colored if you are using the Methylene blue as counterstain The appearance of Background —Green colored if you are using the malachite green as counterstain or Blue colored if you are using the Methylene blue as counterstain REPORTING OF AFB IN ACID FAST STAINING RESULTS For precise and accurate reports of the test, the Acid-fast stained smears are observed at least for 300 High Power Fields HPF before reporting. The center functions as a referral and consulting center with related research and training activities. These appear blue after staining is completed. It is an important differential stain used to identify bacteria in the genus Mycobacterium, some of which are pathogens e.
What makes Mycobacterium resistant to staining? Attach the required drawing illustrating acid fast stained cells. Begin the procedure with the dried slide on the slide warmer. Carbon dioxide CO2 from the atmosphere dissolves to form carbonic acid. Most American patients are treated under U. Waksman went on the win the Nobel Prize for medicine in 1952 and took full credit for the discovery. The Ziehl-Neelson method uses steam heat to allow stain to penetrate, whereas the Kinyoun cold method uses a wetting agent mixed in with the primary stain. By forming spores these bacteria can survive in harsh conditions, spores are resistant to heat, dessication, chemicals and radiation.
Interpretation of Results & Reporting of AFB in Sputum Smears
Moreover, AFB can be transferred from one smear to another when the same piece of blotting paper is used to dry several smears, which is usually avoided as it increases the chances of erroneous results. The carbon atom of a carboxyl group has a high oxidation state. Figure1: Mycobacterium tuberculosis under 1000X oil immersion objective Image source: Microbeonline. Escherichia coli Procedure: 1. Acid fast is used when acid fast positive bacteria are suspected. After making an Acid-fast stained smear of the specimen a Laboratory technologist has to carefully observe under the microscope for the Red-colored Thin, Beaded curved rods which is a characteristic morphology of Mycobacterium tuberculosis and thereafter counting the no. Lab Questions: This procedure is used to separate what two types of bacteria? Microbiology Lab Notebook Report Acid Fast Stain - Exercise 3- March 17, 2020 Introduction The acid-fast stain is a differential stain used to detect cells capable of retaining a primary stain when treated with an acid alcohol.
Air dry and then heat-fix. Are acid-fast negative cells stained by carbolfuchsin? Since we are not using heat, it is important to leave the stain on for a long time to allow it to penetrate the spore coat. The genus Mycobacterium and the genus Nocardia possess an acid-fast cell wall. By heating carbol fuchsin, then methylene blue is added. Several species of Mycobacterium and Nocardia are pathogenic for animals and humans.
Fill the buret with a weak base and place it in the ring stand. Then, place the slide on the staining tray and counterstain the smear s with methylene blue for 1 minute. According to the CDC, it is estimated that 23% of the global population has latent tuberculosis infections. Be sure the staining tray catches any excess stain. Ehrlich observed that dyes were not absorbed by Mycobacterium unless the organism on the slide was heated.