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2420-Lab 3- Bacterial Staining Techniques-II-(Differential) Directions: Answer following questions after reading the information and watching the video from the link below. Use color RED for your answers. Submit the completed document on eCampus for grading. Refer to the textbook Openstax Microbiology Chapter 2 Link: Bacterial Staining Techniques-II, Virtual Edge Experiment-4 This document has 34 questions Students MUST use scientific terminology while answering all questions For each wrong answer 0.5 points will be deducted Differential Staining What is differential staining? Gram Staining (4A on Virtual Edge) Watch the video of Gram Staining from the textbook chapter 3- review 3.2 (pages 53-54), 3.5 (pages 65-67- cell wall of prokaryotic cells) and answer the following questions. What is the purpose of Gram staining? Who invented the Gram staining procedure? What is the first stain (primary stain) used in Gram staining procedure? Do all cells pick up the primary stain? What is the function of a mordant? Which mordant is used in the Gram staining procedure? What would be the result if the mordant is forgotten from the procedure? What is alcohol used for? Why is decolorization necessary? What is a counterstain? Which counterstain (or secondary stain) is used in Gram staining? What are cells called that retain the primary stain? What are cells called that lose the primary stain? What are cells called that take up the counterstain? In the Gram staining process, what type of cells do not take up the counterstain? After the Gram staining procedure, Gram positive bacterial cells appear ______________ colored. After the Gram staining procedure, Gram negative bacterial cells appear ______________ colored. What are the components of the cell wall in Gram positive bacteria? What are the components of the cell wall in Gram negative bacteria? What would be the color of Gram negative bacterial cells if the decolorization step is forgotten? Explain. Which bacteria were used in this experiment and what were their shapes? (From the textbook, review 3.3- Morphology of prokaryotic cells) From what you have learned from the textbook, if a patient has E. coli infection which are Gram negative bacteria, would you advise them to take a course of Penicillin? Why or Why not?- Explain Following picture shows the Gram staining result. Point out (or describe) Gram positive and Gram negative cells from the picture. Be sure to specify morphology: Acid-fast Staining- (4B on Virtual Edge): Read the text from the following to help answering the following questions. link Acid-fast Staining, textbook chapter 3- section 3.2 List two genera of bacteria that are stained for identification by using Acid-fast staining method. Which fatty acid is found in the cell walls of acid-fast bacteria? Which primary stain is used in the acid-fast staining procedure? Why is steaming necessary at this step? Which counterstain is used in the acid-fast staining procedure? Why is acid-alcohol used to decolorize in the acid-fast staining process instead of regular ethanol? Which two bacterial species were talked about in this video? Which of the above two types of bacteria were acid-fast? (or retained the primary stain?) From the picture below, point out the acid-fast cells: If you are working in a clinical diagnostic laboratory, which of the two staining methods you learned in this lab would you use to predict the cause of infection for following patients? A patient with symptoms of Tuberculosis: ______________________________________ staining A patient with symptoms of Strep throat: _______________________________________ staining
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