to do electron microscopy lecture results from environmental plates & broths serial dilution...
TRANSCRIPT
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To Do• Electron Microscopy Lecture
• Results from Environmental Plates & Broths
• Serial Dilution & Streak Plates
• Fungi: make wetmounts & observe demo’s
• Observe gram stain demo’s/finish old slides
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Relative Size of objects
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Resolution Analogy
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Resolution
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Anatomy of a Wave
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Properties of Light
• Wavelength: Length of a light ray; represented by the Greek letter lambda ()
• Equal to the distance between two adjacent crests or troughs of a wave
• Resolution: Refers to the ability to see two items as separate and discrete units
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Properties of Light
• Resolving Power (RP): minimal distance two points can be distinguished as two points
• The smaller the distance between objects that can be distinguished, the greater the resolving power of the lens
• RP= /2(NA)
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Microscope Comparison
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Modern Scanning Electron Microscope
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Limits of Resolution
• Eye: 0.2 mm = 200 microns = 200,000 nm
• BF scope: 0.0002 mm =0.2 microns = 200 nm
• TEM: 0.00001 mm = 0.01 microns = 10 nm
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E . coli Brightfield
KM
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Comparison of Electron Microscopes
E.coli SEM E. coli TEM
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Resolution vs. Magnification
Light Microscope SEM
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SEM
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TEM
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Type of Scope?
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Type of Scope?
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ID Microbe & Specific Scope
CDC, used with permission
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Microscope comparison John Wiley
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Results from Environmental Samples
• Learn how to interpret broth culture growth patterns
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Broth comparison
uninoculated
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Various Colored Colonies
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Review Properties of Agar• Comes from Red algae
• Microbes usually don’t eat it
• Not toxic
• Melts at 100 C
• Solidifies at 40 C
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Preparation of a pure Culture
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4 Quadrant Streak Plate
1. swab
2. loop
3. loop
4. loop
Do right NOW
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Quantitation of Bacteria
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Serial Dilutions
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Comparison of Serial Dilution Techniques
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Pour Plate
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Which of these plates would be the correct one to count? Why?
Countable number of colonies (20 to 200 per plate)
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Serial Dilutions
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Fungal Demo’s Gram Stain Demo’s
• Aspergillus Treponema pallidum
• Rhizopus Neisseria gonorrhoeae
• Penicillium Corynebacterium
• Candida
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Aspergillus
100x 400x
KM
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What are these?
KM400x
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Rhizopus
KM100x
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Penicillium
100x 400x
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Candida
100x 400x
KM
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Candida & staphylococci
KM
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Fungal Demo’s Gram Stain Demo’s
• Aspergillus Treponema pallidum
• Rhizopus Neisseria gonorrhoeae
• Penicillium Corynebacterium
• Candida
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Treponema
CDC, used with permission
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Treponema
1000xKM Wikipedia Commons, used with permission
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Diplococci of Neisseria gonorrhoeae (4087X)
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TEM diplococci of Neisseria gonorrhoeae (96,051X)
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Neisseria
1000x KM
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Neisseria
1000x KM
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Corynebacterium diphtheria
CDC, used with permission
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Corynebacterium diphtheria—the cause of diphtheria
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Corynebacterium
1000x KM
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Fungal Demo’s Gram Stain Demo’s
• Aspergillus Treponema pallidum
• Rhizopus Neisseria gonorrhoeae
• Penicillium Corynebacterium
• Candida
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www.udel.edu
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ID Microbe & Specific Scope