18. True or false: Nocardia is another acid-fast bacterial genus A) True B) False 19. Secondary stain in both acid-fast stain methods is: A) Carbol fuchsin B) Crystal violet C) Methylene blue D) Safranin E) Phenol 20. In the Ziehl-Neelsen acid-fast staining, what is the mordant? A) Carbol fuchsin B) Heat C) Methylene blue D) Basic fuchsin 21. The two common techniques used to visualize endospores: A) Simple staining B) Gram staining C) Negative staining D) Schaeffer-Fulton and crystal violet staining E) All of the above 22. Which of the following statements about endospores is false? A) Some species of bacteria produce metabolically inactive, dormant forms called endospores B) Endospores are part of a reproductive cycle C) Endospores develop within the vegetative bacterial cells D) Endospores are resistant to harsh environmental conditions E) None of the above 23. With negative staining (crystal violet) color of endospores is: A) Purple B) Green C) Red D) Colorless E) None of the above
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Step 2: The secondary stain in both acid-fast stain methods is Methylene blue. Step 3: In the Ziehl-Neelsen acid-fast staining, the mordant is heat. Step 4: The two common techniques used to visualize endospores are Schaeffer-Fulton and crystal violet Show more…
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Special Staining Negative staining - can be done using an acid dye like Nigrosin dye or India ink; the cells don't pick up the stain, clear cells are seen against dark surrounding. -in capsule staining, encapsulated bacteria are mixed with a solution containing Nigrosin dye or India ink, then stain with a simple (basic) stain. Capsule appears as clear halo surrounding bacterial cell. ii. Endospore staining- to stain the endospores What is used as primary stain? __________ What is used as a mordant? __________ What is used as decoloriser? __________ What is used as counterstain? __________ The endospores retain the __________ color of the primary stain __________ The vegetative cells are colored __________, same color of counterstain __________ iii. Flagella staining stain used is: __________
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Gram stain: Color Gram reaction Cell Morphology Endospore stain: Color Endospores present: Yes No Acid Fast stain: Color Acid fast cells: Yes No Complete the table and highlight ALL that are consistent with your unknown. If none are, provide possible explanation. Gram reaction Cell morphology Endospores? Acid fast? Enterococcus faecalis + Cocci - No Staphylococcus aureus + Cocci - No Mycobacterium smegmatis + Bacillus - Yes Escherichia coli - Bacillus - No Bacillus subtilis + Bacillus * No Pseudomonas aeruginosa - Bacillus - No Clostridium sporogenes + Bacillus * No Micrococcus luteus + Cocci - No Corynebacterium xerosis + Bacilli - No Indicate whether the results are positive or negative and the name of the dye resulting in the pink color for each of the following: Positive or negative Dye Pink color after Gram stain Neg. Safranin 0.25% Pink color after endospore stain Neg. Safranin 0.25% Pink color after acid fast stain Positive Carbolfuchsin Bacillus subtilis is a Gram-positive rod that produces endospores and is not acid fast. What color will it be after each of the staining procedures? What dye is responsible for the color? COLOR Dye Gram stain Purple Crystal Violet Endospore stain Green Malachite Green Acid fast stain Blue Methylene Blue
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Acid-Fast Staining: Kinyoun Method A. Results Draw cells from the acid-fast stained slide. Label your drawing to differentiate acid-fast from non-acid-fast cells. M. smegmatis and S. epidermidis (Kinyoun method) B. Short-Answer Questions 1. What makes Mycobacterium resistant to staining? 2. What other bacterial genus is acid-fast? 3. What is the primary stain in the Kinyoun acid-fast stain? How does this differ from the primary stain in the Ziehl-Neelsen method? 4. What is the secondary stain in both acid-fast stain methods? 5. In the Ziehl-Neelsen acid-fast stain, what is the mordant?
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