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  Section: Biotechnology Methods » Microbiology
 
 
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Use of Darkfield Illumination

 
     
 
Content
Microbiology
  Introduction
  The Microscopy
  The Bright Field Microscope
  Introduction to the Microscope and Comparison of Sizes and Shapes of Microorganisms
  Cell Size Measurements: Ocular and Stage Micrometers
  Measuring Depth
  Measuring Area
  Cell Count by Hemocytometer or Measuring Volume
  Measurement of Cell Organelles
  Use of Darkfield Illumination
  The Phase Contrast Microscope
  The Inverted Phase Microscope
  Aseptic Technique and Transfer of Microorganisms
  Control of Microorganisms by using Physical Agents
  Control of Microorganisms by using Disinfectants and Antiseptics
  Control of Microorganisms by using Antimicrobial Chemotherapy
  Isolation of Pure Cultures from a Mixed Population
  Bacterial Staining
  Direct Stain and Indirect Stain
  Gram Stain and Capsule Stain
  Endospore Staining and Bacterial Motility
  Enumeration of Microorganisms
  Biochemical Test for Identification of Bacteria
  Triple Sugar Iron Test
  Starch Hydrolysis Test (II Method)
  Gelatin Hydrolysis Test
  Catalase Test
  Oxidase Test
  IMVIC Test
  Extraction of Bacterial DNA
  Medically Significant Gram–Positive Cocci (GPC)
  Protozoans, Fungi, and Animal Parasites
  The Fungi, Part 1–The Yeasts
  Performance Objectives
  The Fungi, Part 2—The Molds
  Viruses: The Bacteriophages
  Serology, Part 1–Direct Serologic Testing
  Serology, Part 2–Indirect Serologic Testing

Materials
  • Microscope with dark-field stop
  • Suspension of Amoeba proteus
  • Transfer pipette
  • Slides and coverslips.
Procedure
  1. Make a simple wet mount of the amoeba by placing a drop of the culture on a slide and placing a cover slip on the slide.
  2. Observe and measure the amoeba using normal bright field optics.
  3. Place a dark field stop in the filter holder below the microscope’s substage condenser and continue to monitor the movements of the amoeba.
  4. Note the differences in observable structure possible with dark-field microscopy when compared to bright field. Draw and label the amoeba viewed in both ways.

 
     
 
 
     




     
 
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