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Microscopy Techniques for Preserving and Staining Organisms

This informative guide covers fixation methods in microscopy to preserve internal and external structures of organisms. Learn about heat and chemical fixation, as well as light microscopy dyes and staining techniques such as Gram and acid-fast staining. Understand how these techniques help differentiate bacteria and visualize specific cellular structures.

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Microscopy Techniques for Preserving and Staining Organisms

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  1. BIOL 3340

  2. Chapter 2 Microscopy

  3. Fixation • preserves internal and external structures and fixes them in position • organisms usually killed and firmly attached to microscope slide • heat fixation – routine use with procaryotes • preserves overall morphology but not internal structures • chemical fixation – used with larger, more delicate organisms • protects fine cellular substructure and morphology

  4. Light Microscopy

  5. Dyes and Simple Staining Dyes • make internal and external structures of cell more visible by increasing contrast with background • have two common features • divides microorganisms into groups based on their staining properties • e.g., Gram stain • e.g., acid-fast stain

  6. Gram Staining Gram Staining • most widely used differential staining procedure • divides bacteria into two groups based on differences in cell wall structure

  7. Acid-fast Staining • Acid Fast Staining: • particularly useful for staining members of the genus Mycobacterium • e.g., Mycobacterium tuberculosis – causes tuberculosis • e.g., Mycobacterium leprae – causes leprosy • high lipid content (mycolic acids) in cell walls is responsible for their staining characteristics

  8. Gram-negative Cell Walls and Acid Fast Fast cell wall in Chapter 3

  9. Bibliography • http://en.wikipedia.org/wiki/Scientific_method • https://files.kennesaw.edu/faculty/jhendrix/bio3340/home.html • Lecture PowerPoints Prescott’s Principles of Microbiology-Mc Graw Hill Co.

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