Carbol Fuchsin Acc. to Ziehl-Neelsen

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Rev.07– 27/10/2011 Carbol Fuchsin acc. to Ziehl-Neelsen Manufacturer: Diapath Via Savoldini,71- 24057 MARTINENGO- BG- PH+39.0363.986.411 [email protected] CODE PACKAGING C0421 125 ml C0422 500 ml C0423 1000 ml Description Carbol fuchsin solution is used for the staining of acid resistant bacteria in the Ziehl-Neelsen method (see, also special stain kit code 010201). This staining is suitable to highlight mycobacteria, Nocardia and parasites on histological sections, smears, excreta and cultures. The protocol is based on typical structure of acid resistant bacteria, which acquire and keep dyes so that following decolorizing treatments are possible. Dark blue background is obtained with Methylene blue. Composition Phenol CAS No. 108-95-2 EC No. 203-6327 Basic fuchsine CAS No. 58969-01-0 EC No. 221-816-5 C.I. 42510 Ethanol CAS No.64-17-5 EC No.20-578-6 Staining protocol Histological sections (Ziehl-Neelsen staining) 1. Dewax sections and hydrate to distilled water 2. Carbol fuchsin solution for 30 minutes 3. Wash very well in running cold water 4. Acid alcohol* till sections are pale pink 5. Water for 5 minutes 6. Methylene blue solution for 30 seconds 7. Distilled water 8. Dehydrate very quickly, clarify and mount *Acid alcohol: 100 ml of ethyl alcohol 70° + 3 ml hydrochloric acid Cytological specimens (Ziehl-Neelsen staining) 1. Carbol fuchsin for 30 minutes 2. Wash in distilled water 3. Acid alcohol* 10 seconds 4. Wash in distilled water 5. Methylene blue for 30 seconds 6. Distilled water 7. Dehydrate very quickly, clarify and mount NOTE: Methylene blue could cover the possible presence of acid resistant bacteria in the specimen. Results Acid resistant bacilli: bright Red Other bacteria and tissues (background contrast): Blue Erythrocytes: Yellow 1 Rev.07– 27/10/2011 Storage, handling and stability Store at room temperature. Keep containers tightly close. Keep away from sources of ignition. No smoking. The product if opportunely stored and integrally packed, is stable up to the expiry date reported on the label. MSDS available for further information. Professional use. Bibliography “Laboratory methods in histotechnology”-Edna B.Prophet ,B Mills,J. Arrigton, L. Sobin-American Registry of Pathology. “Theory and practice of Histological Techniques” J.D. Bancroft; M. Gamble. Churchill Livingstone- 5° edition 2002 2 .

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CARBOL FUCHSIN STAIN (ZIEHL-NEELSEN) - for in Vitro Use Only - Catalogue No
CARBOL FUCHSIN STAIN (ZIEHL-NEELSEN) - For in vitro use only - Catalogue No. SC24K Our Carbol Fuchsin (Ziehl-Neelsen) Stain is Formulation per 100 mL used in the microscopic detection of acid-fast microorganisms such as Mycobacterium . SC25 Carbol Fuchsin Stain (Ziehl-Zeelsen) Acid-fast organisms such as Mycobacterium Basic Fuchsin ..................................................... 0.3 g have cell walls that are resistant to conventional Phenol ................................................................ 5.0 g staining by aniline dyes such as the Gram stain. Ethanol ............................................................ 10 mL However methods that promote the uptake of dyes De-ionized Water ............................................. 90 mL are available; once stained these organisms are not easily decolorized even with acid-alcohol or acid- SC26 Carbol Fuchsin Decolorizer acetone solutions therefore they are described as Hydrochloric Acid .......................................... 3.0 mL acid-fast. Their resistance to destaining is a useful Ethanol .......................................................... 97.0 mL characteristic in differentiating these organisms from contaminating organisms and host cells. SC27 Carbol Fuchsin Counterstain (Methylene Blue) The Ziehl-Neelsen staining procedure is often Methylene Blue ................................................. 0.3 g referred to as hot carbolfuchsin because of the need De-ionized Water ............................................100 mL to apply heat during the staining
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SURGICAL PATHOLOGY - HISTOLOGY Date: STAINING MANUAL - MICROORGANISMS Page: 1 of 3 ACID-FAST BACTERIA - ZIEHL-NEELSEN STAIN (AFB) PURPOSE: Used in the demonstration of acid-fast bacteria belonging to the genus 'mycobacterium', which include the causative agent for tuberculosis. PRINCIPLE: The lipoid capsule of the acid-fast organism takes up carbol- fuchsin and resists decolorization with a dilute acid rinse. The lipoid capsule of the mycobacteria is of such high molecular weight that it is waxy at room temperature and successful penetration by the aqueous- based staining solutions (such as Gram's) is prevented. CONTROL: Any tissue containing acid-fast organisms. Use Millipore™ filtered water in the waterbath and staining procedure. FIXATIVE: 10% formalin TECHNIQUE: Cut paraffin sections at 4-5 microns. EQUIPMENT: Rinse all glassware in DI water, coplin jars, microwave oven. REAGENTS: Ziehl-Neelsen Carbol-Fuchsin 1% Acid Alcohol: Solution: Hydrochloric acid 10.0 ml Basic fuchsin 2.5 gm 70% Alcohol 990.0 ml Distilled water 250.0 ml Mix well, label with date and 100% alcohol 25.0 ml initials, stable for 1 year. Phenol crystals, melted 12.5 ml CAUTION: Corrosive. Mix well, filter into brown bottle. Label bottle with date and initials, solution is stable for 1 year. CAUTION: Carcinogen, toxin. MICROOGANISMS ZIEHL-NEELSEN-AFB Page: 2 of 3 Methylene Blue Methylene Blue Stock Solution: Working Solution: Methylene blue 0.7 gm Methylene Blue Stock 5.0 ml Distilled water 50.0 ml Distilled water 45.0 ml Mix well, filter into bottle. Label Pour into coplin jar, stable for 2 with date and initials, stable for 1 months.
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Carbol Fuchsin Ziehl Neelsen.1030
2505 Parview Road ● Middleton, WI 53562-2579 ● 800-383-7799 ● www.newcomersupply.com ● [email protected] Part 1030 Revised July 2019 Carbol Fuchsin Stain, Ziehl-Neelsen for AFB Stain - Technical Memo SOLUTION: 250 ml 500 ml 1 Liter Carbol Fuchsin Stain, Ziehl-Neelsen Part 1030A Part 1030B Part 1030C Additionally Needed For AFB Stain, Ziehl-Neelsen: Acid Fast Bacteria (AFB) Control Slides Part 4011 Acid Alcohol 1% Part 10011 Light Green SF Yellowish Stain 0.1%, Aqueous Part 12203 or Methylene Blue Stain 0.14%, Alcoholic Part 12401 Xylene, ACS Part 1445 Alcohol, Ethyl Denatured, 100% Part 10841 Alcohol, Ethyl Denatured, 95% Part 10842 For storage requirements and expiration date refer to individual product labels. APPLICATION: RESULTS: Newcomer Supply Carbol Fuchsin Stain, Ziehl-Neelsen, a crucial Acid-fast bacilli Bright red element in the AFB Stain, Ziehl-Neelsen is used to demonstrate the Background Green (with Light Green SF Yellowish counterstain) presence of acid-fast mycobacteria in tissue sections. Acid-fastness is a Background Pale blue (with Methylene Blue counterstain) physical property of certain bacteria and cellular structures. Carbol Fuchsin Stain, Ziehl-Neelsen, combines phenol and basic fuchsin that PROCEDURE NOTES: works to permeate the lipoid capsule of acid-fast organisms and renders them resistant to acid alcohol decolorization. 1. Drain slides after each step to prevent solution carry over. 2. Do not allow sections to dry out at any point during procedure. METHOD: 3. Sections can remain in Carbol Fuchsin Stain, Ziehl-Neelsen for up to 60 minutes without adverse effect. Additional differentiation may Fixation: Formalin 10%, Phosphate Buffered (Part 1090) be required in Step #6.
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