The omitted step in a Gram stain method that allow differentiation of Gram-positive and Gram-negative cells. Given: “Gram stain” “differentiation “and “ omitted step ” Introduction: In 1884, the Danish scientist Hans Christian Joachim Gram is discovered the Gram staining procedure to differentiate the bacteria into two groups, Gram-positive and Gram-negative. It is the preliminary step to identify the staining characteristics of bacteria based on the morphology and arrangement, which is observed under the light microscope. The main principle of Gram stain is used to differentiate based on the color retaining ability of the stain. Gram-positive bacteria retains crystal violet and appears violet or purple in color under the microscope. Since, the cell wall of Gram-positive bacteria contain thicker peptidoglycan layer. So, it retains the stain during ethanol decolonization step. Whereas in the Gram-negative with thinner peptidoglycan layer denatured during ethanol step and retains the secondary stain (safranin) and appears pink color under the microscopic examination.

Question

Describe in detail all the steps needed to determine whether an Unknown bacteria has a Gram-positive or Gram-negative cell wall using the Gram stain. (You can describe the method presented in the Exercise 3-6 Gram stain videos or in the Lab Manual.) 1. Start by describing the method of transferring the bacteria onto the microscope slide, then describe heat-fixing the emulsion of the Unknown bacteria. (Do not describe all the steps needed to perform aseptic technique. Just say “Bacteria was added aseptically.”) 2. Describe the Gram stain in detail. (This description should include all the stains, how long the stain should set on slide, and Genus and species names of bacteria you are using as CONTROLS.)

Answer 1

Question

Chapter 3, Problem 1A

Summary Introduction

To write:

The omitted step in a Gram stain method that allow differentiation of Gram-positive and Gram-negative cells.

Given:

“Gram stain” “differentiation “and “omitted step”

Introduction:

In 1884, the Danish scientist Hans Christian Joachim Gramis discovered the Gram staining procedure to differentiate the bacteria into two groups, Gram-positive and Gram-negative. It is the preliminary step to identify the staining characteristics of bacteria based on the morphology and arrangement, which is observed under the light microscope.

The main principle of Gram stain is used to differentiate based on the color retaining ability of the stain. Gram-positive bacteria retains crystal violet and appears violet or purple in color under the microscope. Since, the cell wall of Gram-positive bacteria contain thicker peptidoglycan layer. So, it retains the stain during ethanol decolonization step. Whereas in the Gram-negative with thinner peptidoglycan layer denatured during ethanol step and retains the secondary stain (safranin) and appears pink color under the microscopic examination.

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Describe in detail all the steps needed to determine whether an Unknown bacteria has a Gram-positive or Gram-negative cell wall using the Gram stain. (You can describe the method presented in the Exercise 3-6 Gram stain videos or in the Lab Manual.) 1. Start by describing the method of transferring the bacteria onto the microscope slide, then describe heat-fixing the emulsion of the Unknown bacteria. (Do not describe all the steps needed to perform aseptic technique. Just say “Bacteria was added aseptically.”) 2. Describe the Gram stain in detail. (This description should include all the stains, how long the stain should set on slide, and Genus and species names of bacteria you are using as CONTROLS.)

2. Explain the relationship between bacterial cell wall with gram positive and gram negative in the staining process. Please state the chemicals used and their functions.

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