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CH 3: Observing Microorganisms Through A Microscope
CH 3: Observing Microorganisms Through A Microscope
CH 3: Observing Microorganisms Through A Microscope
Observing
Microorganisms
Through a
Microscope
Q&A
Acid-fast staining of a
patient’s sputum is a
rapid, reliable, and
inexpensive method to
diagnose tuberculosis.
What color would
bacterial cells appear if
the patient has
tuberculosis?
Objectives
Review the metric units of measurement
Define total magnification and resolution
Explain how electron and light microscopy differ
Differentiate between acidic and basic dyes
Compare simple, differential, and special stains
List the steps in preparing a Gram stain. Describe the
appearance of Gram-positive and Gram-negative
cells after each step
Compare and contrast Gram stain and acid-fast stain
Explain why endospore and capsule stains are used
Units of Measurement
• 1 µm = ______ m = ______ mm
• 1 nm = ______ m = ______ mm
• 1000 nm = ______ µm
• 0.001 µm = ______ nm
Sizes Among Microorganisms
• Protozoa: 100 µm
Cells Alive –
• Yeasts: 8 µm How big is a . . .?
Fig 3.6b
Principle of
Immunofluorescence
Electron Microscopy: Detailed Images of
Cell Parts
Uses electrons, electromagnetic lenses, and
fluorescent screens
Electron wavelength ~ 100,000 x smaller than
visible light wavelength
Specimens may be stained with heavy metal
salts
Two types of EMs:?
SEM or TEM?
Bacterial division
Leaf surface
?
crystal violet
safranin
Acid Fast Stain
• Cells that retain a basic stain in the presence of
acid-alcohol are called acid-fast.
• Non–acid-fast cells lose the primary stain when
rinsed with acid-alcohol, and are counterstained
with a different color basic stain
Special Stains See Fig 3.14