Bacillus subtilis (Gram-positive) and Pseudomonas aeroginosa (Gram-negative) bacteria are heat-fixed on slide and subjected to Gram staining. What do you observe under the microscope when the following steps are skipped? Please explain. a. Staining with safranin b. Staining with Gram iodine c. Decolorization

Bacillus subtilis (Gram-positive) and Pseudomonas aeroginosa (Gram-negative) bacteria are heat-fixed on slide and subjected to Gram staining. What do you observe under the microscope when the following steps are skipped? Please explain. a. Staining with safranin b. Staining with Gram iodine c. Decolorization

Basic Clinical Laboratory Techniques 6E

6th Edition

ISBN:9781133893943

Author:ESTRIDGE

Publisher:ESTRIDGE

Chapter7: Basic Clinical Microbiology

Section7.7: Bacterial Identification And Antibiotic Susceptibility Testing

Problem 2CS

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Explain why only gram-negative cells undergo decolorization during the gram staining procedure. Cite the purpose of each of the following reagents in a differential staining procedure. Primary stain Mordant Decolorizing Agent Counter stain What might happen if the Gram staining procedure is performed on a culture incubated for a little over a day
Describe the procedure for performing a Gram stain. What is the procedure? How does each component work? (ie. What does the mordant do?)
The acid-fast bacterium, Mycobacterium smegmatis, is relatively safe to be used by students in performing the acid-fast stain technique because of its thin cell wall. However, if this organism is used for Gram-stain, it can also take the Gram stain reaction. Why is this so?
Based from the results obtained from Gram staining, explain why bacterial cells retained or changed their color. Briefly explain the principle behind the procedure. Cite advantages of differential staining method (Gram staining and endospore staining) over simple staining only. List other staining methods aside from those you performed in the laboratory. conclusion:
Which of the following is/are true regarding the acid-fast stain? (There may be more than one correct answer.) Non-acid-fast microbes appear blue in a completed acid-fast stain. O Acid-fast cells retain the primary dye after treatment with acid-alcohol. O It is used to identify members of the genus Mycobacterium. V Acid-fast cells appear bright pink/red in a completed acid-fast stain. O If cells are acid-fast, they are gram-negative.
a. Identify the cell shape, cell arrangement, and Gram reaction of the Unknown microbe in the image below. b. Suppose the corresponding Gram negative control were purple. Explain what step(s) of the Gram stain you would adjust and how you would adjust them to ensure the accuracy of your Unknown results
a. What can you observe in viewing your stained bacterial slide under the microscope if you fixed a lot of bacterial colonies in your slide during smear preparation? b. What can you observe in viewing your stained bacterial slide under the microscope if you heat fixed your slide in a much longer exposure to heat?
In Gram staining method, list the reagents and state their role for each step of the procedure? In preparing bacterial smear, why do we need to pass the slide over the flame?
The bacterial cells in the image below were stained with a high concentration of carbolfuschin for an extended period of time (5 minutes) while the slide was heated. The slide was decolorized with a mixture of acid and alcohol. The slide was then counter-stained with methylene blue. What kind of stain does this describe?
A mixed culture is known to contain Gram negative bacilli and Gram positive cocci. You perform a Gram stain of this mixed culture as a control while simultaneously Gram stainin an unknown organism on the same slide (the same procedure you used in lab). The results, observed under the 100x objective lens, are described below: Results: Control side: purple bacilli and purple cocci Unknown side: purple bacilli Using all the information given above, discuss the results of your unknown organism. Shoul you be confident in this result? Answer question A if you are confident, answer question B if you are not confident. a. If you are confident in your results, why? Explain how the staining reagents produced the described results. b. If you are not confident, do you think an error was made in the Gram staining procedure? If so, describe the error and explain how it impacted your results.
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Why is it important to perform a gram stain for any type of bacteria? After performing the gram stain method what would be the next step? (Think in terms of the real-world situation) Explain.