In a Gram-stained smear, medium-sized rods with rounded, pink ends were found, arranged chaotically. 1. What are the tinctorial properties of the bacteria from which the smear is made? 2. What is the purpose of studying the tinctorial properties of microorganisms? 3. How to stain the slide by the Gram method?
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- 1. Why is it important to use a small amount of bacteria when making a smear? Give at least 2 reasons. 2. What are the two reasons for “fixing” a smear? 3. Simple staining is useful for determining what three characteristics of a bacterial cell? 4. With respect to bacteria, define morphological arrangement. Explain thoroughly!1.why is gram stain considered a differential stain? 2.How do gram positive and gram negative bacteria differ in cellular structure, and how does this contribute to their differential staining properties? 3.How does the age of a culture affect the gram stain reaction? What is an optimum culture age for a valid gram reaction? 4.Which step in the gram stain procedure is most prone to error?If done correctly how might that step affect the end result? 5.what is the function of mordant, and which reagent serves this purpose in the gram stain procedure? 6.List the reagents of the gram star technique in order and their general role in the staining process. 7.In what type of cell, gram -positive or gram-negative , would you find lipopolysaccharide in its cell wall?1. What is the purpose for using stains? What microbial characteristics can one ascertain from a simple stain? 2. Why is it necessary to make a heat-fix smear and what are the disadvantages of heat fixing? 3. what is the best age for your culture when performing a gram stain? Why? 4. Why is Gean staining considered a differential staining process? 5. What are some reasons a Gram positive cell might appear Gram negative?
- 1. What color does an acid-fast cell stain? 2. Identify two diseases that are caused by acid-fast bacteria. 3. In the endospore stain, what color do the endospores stain? 4. If you performed an endospore stain on Mycobacterium, what color cells would you expect tosee? Why? 5. How do capsules contribute to virulence of an organism? 6. Since you know what lophotrichous and amphitrichous arrangements look like, put thoseterms together to draw an amphilophotrichous bacterium. 7. Streptococcus pneumoniae that are capable of causing pneumonia are encapsulated bacteria(meaning they have a capsule). Describe what you would expect to see under themicroscope after performing a capsule stain with india ink and safranin.1. What are the advantages of staining a bacterial preparation before observing it under a microscope?2.Briefly state how a hanging-drop preparation is prepared.3. How does the heaviness of a bacterial smear affect its microscopic analysis4. Why should you be careful not to underheat a smear during the heat-fixing process5.What is heat fixation? How is it carried out?6. Why do you think the presence of grease or dirt on a glass slide will result in a poor smear preparation? Cite two or three reasons 7. Why are basic dyes more effective for bacterial staining than acidic dyes?8. State two ways that can confirm whether a bacterial smear has been correctly prepared or not1) In Gram staining, to what cell structure do the dyes bind? 2) Would it be useful to perform a Gram stain on a mixed culture? Why? 3) In capsule staining, why does the capsules did NOT take in any dye? 4) In endospore staining, what is the purpose of using the steam?
- 1.)What is the purpose of a counterstain? 2. What does a mordant do in the Gram stain procedure? Which reagent in the Gram stain is the mordant? 3. True or False? The oil objective should make contact with the oil on the slide. 4. Why is it necessary to let bacterial smears completely air dry before heat fixing? 5.Why should controls be included wherever possible for any staining technique? 6. Why is it necessary to heat the slides while staining for endospores?1. You accidentally used safranin as the primary stain and malachite green as the counter stain during a spore staining procedure. Explain how your microscopic observation would differ from those observed when slides of spore-containing bacteria were prepared correctly. 2.One of your classmates performed a gram stain on Pseudomonas aeruginosa and found variable gram staining, with only some of the cells gram positive. What should your classmate have found? What errors could have contributed to the variable result?How would you expect the staining properties of 24-hour culture of Bacillus subtilis or the other Gram-positive bacteria to compare to culture that is 3 to 4 days older?
- Are there any chemical differences between the cell walls of gram-positive and negative bacteria that might explain differences in the rate of decolorization ? Does the age of a culture affect the gram stain reaction ? Why or why not?2.How do gram positive and gram negative bacteria differ in cellular structure, and how does this contribute to their differential staining properties? 3.How does the age of a culture affect the gram stain reaction? What is an optimum culture age for a valid gram reaction? 4.Which step in the gram stain procedure is most prone to error?If done correctly how might that step affect the end result? 5.what is the function of mordant, and which reagent serves this purpose in the gram stain procedure? 6.List the reagents of the gram star technique in order and their general role in the staining process. 7.In what type of cell, gram -positive or gram-negative , would you find lipopolysaccharide in its cell wall?Answer this question xi) What is the most important step in the Gram staining procedure? Why?