Tell your researcher to come join us in the twentyfirst century. When counterstain is applied methylene blue, nonacidfast bacteria pick it up and stain. In the hot zn technique, the phenol carbol fuchsin stain is heated to enable the dye to penetrate the waxy mycobacterial cell wall. A detailed explanation of the recommended use of 1% hot carbol fuchsin was also. In the ziehlneelsen stain, a fixed smear covered with carbolfuchsin is heated, rinsed, decolorized with acidalcohol, and counterstained with methylene blue. Carbol fuchsin solution, 100ml the curated chemical collection.
Pdf comparison of variants of carbolfuchsin solution in ziehl. Material safety data sheet carbol fuchsin solution 7 us state. Carbol fuchsin is used as the primary stain dye to detect acidfast bacteria because it is more soluble in the cells wall lipids than in the acid alcohol. Section 1 chemical product and company identification. Flammable liquid and vapour harmful if swallowed harmful in contact with skin harmful if inhaled suspected of causing cancer. For use with application used to stain smears prepared from specimens and cultures suspected of containing mycobacteria for early presumptive diagnosis of mycobacterial infection and characterization of bacterial isolates. Heat for 5 min, not allowing the staining solution to boil. In solution with phenol also called carbolic acid as an accentuator it is called carbol fuchsin and is used for the ziehlneelsen and other similar acidfast staining of the mycobacteria which cause tuberculosis, leprosy etc. The carbol fuchsin stain allows you to easily distinguish between bacillus, coccus, and spirillum bacteria.
Flood smear with carbol fuchsin stain zcarbol fuchsin is a lipid soluble, phenolic compound, which is able to penetrate the cell wall 3. Two types of stains are used specifically for detection of mycobacteria. Maintain steaming for 5 minutes by using low or intermittent heat. Page 36 safety data sheet according to safe work australia printing date 31. Acidfast stain principle, procedure, interpretation and. The gram stain kit microorganism stain is intended for the demonstration and differentiation of grampositive and gramnegative bacteria.
Allow the heated stain to remain on the slide for 5 minutes. To prepare carbol fuchsin stain ziehls nielsen, add 1 g of basic fuchsin to 10 ml of absolute ethyl alcohol. Acidfast, spores, and capsule stains biology libretexts. Central drug house p ltd 728 vardaan house new delhi2 india. Carbol fuchsin is used as the primary stain dye to detect acidfast. Primary stain, stains both grampositive and gramnegative cells purple. Carbol fuschin stain definition of carbol fuschin stain by. Carbolfuchsin solution according to kinyoun for microscopy. Castellani paint modified drug information, professional. It can also be used for the staining of bacterial spores. It is commonly used in the staining of mycobacteria as it has an affinity for the mycolic acids found in their cell membranes.
Basic fuchsine is widely used in biology to stain the nucleus, and is also a component of lactofuchsin, used for. Prolab diagnostics dilute carbol fuchsin stain acid fast stains for mycobacteria. The name comes from the danish bacteriologist hans christian gram, who developed the technique gram staining differentiates bacteria by the chemical and physical properties of their cell walls. Acidfast stain principle, procedure, interpretation and examples. Mycobacterium tuberculosis part 4 afb stain acid fast. Oct 27, 2014 register now and get a free online msds binder. It is a component of ziehlneelsen stain, a differential stain. Other companies are charging thousands of dollars to set up accounts and give you access to their msds online database. It is mainly required for the staining of mycobacteria. The ziehl neelsen zn method of staining acid fast bacilli has been in vogue for more than hundred years. Carbol fuchsin definition of carbol fuchsin by medical. Occasionally filter carbol fuchsin if precipitate forms.
Carbol fuchsin is the primary stain in this procedure, and it contains phenol to help solubilize the cell wall. Learning about the typical forms of bacterial cells is an essential lesson. Gram stain or gram staining, also called grams method, is a method of staining used to distinguish and classify bacterial species into two large groups. From our beginnings in 1927, carolina biological supply company has grown to become a leading supplier of science teaching materials for all levels of education. When counterstain is applied methylene blue, nonacidfast bacteria pick it up and stain blue. Mycobacteria and nocardia medical microbiology ncbi. Your new online msds binder is a place for you to store the material safety data sheets you need to deploy. Other products for staining tissue sections find more kits and reagents in the special stains guide, or products for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and.
Safranin and dilute carbolfuchsin are commonly used counterstain in gram staining procedure, another being neutral red it stains gonococci and meningococci well preparation of safranin. Acid fast bacteria will be red, while nonacid fast bacteria will stain bluegreen with the counterstain with the kinyoun stain. Afb stain or ziehlneelsen acidfast staining procedure. Procedure has been corrected from the book of monica chessebrough. The kinyoun stain is modified to make heating unnecessary. Mycobacteria resist this decolorization and are considered acid fast. Acid fast stains are used to differentiate acid fast organisms such mycobacteria. Preparation of gram stain reagents learn microbiology online.
Carbol fuchsin dilute solution zi ehl neelsen staining soln. Thermal decomposition can lead to release of irritating gases and vapors. Then smear from sputum is fixed by heat or alcohol. Add the remaining quantity of absolute ethanol and mix well the contents. Flood the entire slide with carbol fuchsin stain ziehlzeelsen. Acid fastness is a phenomenon that is found in certain types of bacteria where they resist. Carbol fuchsin zn, dilute is used as solution for monochrome staining of bacteria. In smears stained with carbol fuchsin, mycobacteria typically appear as red rods 110. This stain uses carbol fuchsin to stain the lipid walls of acid fast organisms such as m. Ar306 carbol fuchsin artisan gram yellow stain kit 65 ml ar362 carbol fuchsin artisan acidfast bacteria afb light green stain kit 65ml reference number. The slide is flooded with carbol fuchsin, which is then heated to dry and rinsed off in tap water. Physical and chemical properties molecular formula c20h20n3cl. Microscopic examination of tissues or of sputum stained by the acidfast staining procedure is an aid in the diagnosis of tuberculosis.
Basic fuchsin can be found on the following state righttoknow lists. Carbol fuchsin is indicated only during the acute and chronic stages of dermatophytoses. Material safety data sheet carbol fuchsin, strong stain. Ziehlneelsen staining is a bacteriological stain used to identify acidfast organisms, mainly mycobacteria.
A modification of this stain is known as the fite stain and has a weaker acid for supposedly more delicate m. It is named for two german doctors who modified the stain. However, because of disadvantages like controlled heating and nonavailability of spirit for lamps there have been attempts. Bacillus megaterium bacillus, micrococcus luteum coccus, and rhodospirillum rubrum spirillum are shown. Ziehlneelsen staining is a type of acidfast stain, first introduced by paul ehrlich. With this mixed smear slide stained with carbol fuchsin, you can easily view the characteristic size and shape of bacillus, coccus, and spirillum bacteria. Empty containers retain product residue, liquid andor vapor, and can be dangerous. Procedure for the preparation of 1% carbol fuchsin. Comparison of two different strengths of carbol fuchsin in. This is further assisted by the addition of heat in the form of heat steam. This product does not contain any chemicals known to the state of california to cause birth defects or other reproductive harm. Allow the slide to cool briefly and rinse the slide with water.
Three acid fast stain solutions for preparing microscope bacteria slides. Carbol fuchsin is first applied in the acidfast method, which stains every cell. The most commonly used method is the ziehlneelsen method, though there is also a kinyouns method. Add the 1 gram of carbol fuchsin to the flask and mix well by swirling the flask.
Use sparkproof tools and explosion proof equipment. Product name carbol fuchsin strong stain other names carbol fuchsin stain product code acf500 u. Cover the slide with strong ziehlneelsens carbol fuchsin and heat the underside of the slide with a lighted alcoholsoaked swab. Great care must be taken when heating the carbol fuchsin especially if staining is carried out over a tray or other container in which highly fiammable chemicals have collected. Weigh accurately 10 gram of carbol fuchsin on a weighing scale or analytical balance. Heat the slide using a bunsen burner or electric staining rack until it is steaming. For example, with stains like carbol fuchsin, it is necessary that they be kept from light, which can disintegrate them. It is commonly used in the staining of mycobacteria as it has an affinity for the mycolic acids found in their cell membranes it is a component of ziehlneelsen stain, a differential stain. Bacteria types slide, separate smears, carbolfuchsin. The term acid fast is used since they resist decolourization with acid alcohol. Carbol fuchsin is a mix of basic fuchsin and phenol which can be used in procedures for bacterial staining to detect acid fast bacteria. The zielnehlson method employs carbol fuchsin, acid alcohol and a blue or green counter stain. Deionized water glycerine ethyl alcohol phenol basic fuchsin dye methyl alcohol isopropyl alcohol percent 7375 910 710 o o einecs elincs 2317912 2002895 2005786.
If the bacteria is acidfast the bacteria will retain the initial red color of the dye because they are able to resist the destaining by acid alcohol 0. Methylene blue chloride, 1% alcohol solution un1219 carbol fuchsin solution un1992 hydrochloric acid, 1% alcohol solution un2924. Ground and bond containers when transferring material. The technique is also referred to as acid fast staining. California, new jersey, florida, pennsylvania, minnesota, massachusetts. Steam helps to loosen up the waxy layer and promotes entry of the primary stain inside the cell. First we would like to applaud the authors for providing a study on an important test method, the ziehlneelsen afb staining procedure, which receives little attention relative to its critical importance in worldwide tuberculosis control.
The slide is then flooded with a 1% solution of hydrochloric acid in isopropyl alcohol or methanol to remove the carbol fuchsin, thus removing the stain from cells that are unprotected by a waxy lipid layer. Ungraded products supplied by spectrum are indicative of a grade suitable for general industrial use. The kit includes safey data sheets and one 30 ml dropper bottle for each of the followings. With a staining kit therefore, it is much easier and safer to stain specimens by following the instruction manual. Basic fuchsin 1 g absolute ethanol 10 ml phenol crystal 5 g distilled water up to 100 ml instructions for preparation of 1% carbol fuchsin. Special hazards arising from the substance or mixture. Ethyl alcohol can be found on the following state righttoknow lists. Acid fast bacteria have a high content of mycolic acids in their cell walls. To evaluate ziehlneelsen zn staining using variants of carbol fuchsin solution, duplicate smears from 416 samples were stained with zn, one set with 1% basic fuchsin and the other 0. Keep the slide hot and replenish the stain if necessary, taking care not to allow the smear to become dry. According to the book, the mechanism of the acidfast stain is a result of two main things. After destaining with acid alcohol, only nonacidfast bacteria destain since they dont have the thick, waxy lipid covering as acidfast bacteria do. Carbol fuchsin, carbolfuchsin, or carbolfuchsin, is a mixture of phenol and basic fuchsin, used in bacterial staining procedures.
Carbol fuchsin staining solution material safety data. Use of the information, documents and data from the echa website is subject to the terms and conditions of this legal notice, and subject to other binding limitations provided for under applicable law, the information, documents and data made available on the echa website may be reproduced, distributed andor used, totally or in part, for noncommercial purposes provided that echa is. The primary stain used in acidfast staining, carbol fuchsin, is lipidsoluble and contains phenol, which helps the stain penetrate the cell wall. International union against tuberculosis and lung disease. Carbol fuchsin dilute solution material safety data. In a staining kit, all the stains are kept in the right conditions. Harleco diluted solution of carbol fuchsin used as a counterstain in various staining modifications of the gram stain. Store the reagent upto 6 months from preparation date. Pdf comparison of variants of carbolfuchsin solution in. Basic fuchsine is widely used in biology to stain the nucleus, and is also a component of lactofuchsin, used for lactofuchsin mounting. Page 27 safety data sheet according to safe work australia printing date 15. The acid fast bacteria have a waxy substance mycolic acid on their cell wall that makes them impermeable to staining procedures.
The waxy walls of mycobacteria helps in retaining the dye within cells. Prolab diagnostics dilute carbol fuchsin stain is designed to be ready to use. When the smear is stained with carbol fuchsin, it solubilizes the lipoidal material present in the mycobacterial cell wall but by the application of heat, carbol fuchsin further penetrates through lipoidal wall and enters into cytoplasm. Carbol fuchsin is sometimes substituted for safranin since it more intensely stains anaerobic bacteria, but it is less commonly used as a counterstain. Carbol fuchsin, the primary stain contains phenol, which helps solubilize the. Cell staining in microscopy types, techniques, preparations. Heat is also applied during the primary stain to increase stain penetration. The organisms appear as slightly bent, beaded rods 2 to 4 m long and 0. In the zn method, the basic fuchsin phenol dye is used hot there by melting the unsaponifiable waxy substance on the surface of the cell wall. Carbol fuschin stain article about carbol fuschin stain by. In the cold technique known as kinyoun method, stain are not heated but the penetration is achieved by increasing concentration of basic fuchsin and phenol and incorporating a wetting agent chemical. Safety data sheet according to 29cfr19101200 and ghs rev. Carbol fuchsin is a basic dye which forms a yellowbrown compound in presence of acid. Carbol fuchsin zn,strongis used as staining solution for acid fast staining.
This being the case, explain how they end up staining grampositive and gramnegative cells differently in the gram stain. The visualisation of acidfast bacilli afb by the zn staining method is. Composition principle and interpretation mycobacteria afbacid fast bacteria are difficult to stain due to high lipid and wax content in their cell walls. In the ziehlneelsen stain, a fixed smear covered with carbol fuchsin is heated, rinsed, decolorized with acidalcohol, and counterstained with methylene blue. Steam helps to loosen up the waxy layer and promotes entry of the primary stain. Using a digital balance weigh out 1 g of basic fuchsin in a sterile 100 ml flask. Gram staining is used to determine gram status to classify bacteria broadly. Decolorization involves application of alcohol which causes the dehydration of the peptidoglycan of grampositive cells to make it more permeable to the crystal violetiodine complexes. Microscope slide with 3 separate smears showing the characteristic size and form of the following bacteria. Our carbol fuchsin stain, decolorizer and counterstain should be stored at room temperature and protected from light. Acid fast staining of mycobacteria in the form of beadings is obtained by means of a carbolfuchsin solution ziehlneelsen stain prepared from pararosaniline or from certain kinds of basic fuchsin. Under these conditions they have a shelf life of 52 weeks from the date of manufacture. These bacteria are termed acidfast because they are able to resist decolorization with acid alcohol.