PREPARING A SMEAR AND SIMPLE STAIN

PREPARING A SMEAR AND SIMPLE STAIN

1. Materials

   • (1) TSB of Escherichia coli
   • (1) TSA slant of Staphylococcus epidermidis
   • Stain rack
   • Bibulous paper
   • Reverse osmosis water
   • Lens paper
   • (2) Microscope slides
   • Inoculating loop
   • Inoculating needle
   • Clothes pin
   • Bunsen burner
   • Striker
   • Grease pencil
   • Methylene blue stain
   • Immersion oil

2. Objectives

   • Prepare and stain smear bacterial
   • Evaluate the morphology

3. Introduction

   • The smear and stain bacterial permit better observation of the morphology cell
   • Use reactive as Alanine derivative coal tar derivate of benzene.
   • The dye adheres to the bacteria and morphology (shape, arrangement, structure, size) can be more readily seen.

4. Morphology of Bacteria

5. The Chemistry of Staining

   • Staining is based on the principle of opposing charges. The dye used in microbiology is a salt containing two ions
     • Cation (positive charge)
     • Anion (negative charge)
       • Methylene blue stain
       • Violet crystal
       • safranine

6. Prep 1 - Preparing Smears from Broth Cultures

7. Preparing Stains

   • new slide sheet should be used
   • spread a thin film of bacteria on the slide.
   • The smear must be thin

8. Prep 2 - Preparing Smears from Solid Media

9. Procedure 1 - Simple Staining

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