LESSON 5. LLight Microscopy BBright-field microscopy PPhase Contrast microscopy DDifferential...
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Transcript of LESSON 5. LLight Microscopy BBright-field microscopy PPhase Contrast microscopy DDifferential...
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ELECTRON MI-CROSCOPY
LESSON 5
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TYPES OF MICROSCOPY
Light Microscopy Bright-field microscopy Phase Contrast microscopy Differential Interference Contrast (or Nomarski) Dark-field microscopy Fluorescence microscopy Confocal Scanning microscopy
Electron Microscopy Transmission electron microscope (TEM) Scanning electron microscope (SEM)
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Learning Resources
1. Junqueira’s Basic Histology ; pp 7 & 9
2. Looking at the Structure of Cells in the Microscope
http://www.ncbi.nlm.nih.gov/books/NBK26880/#A1749▪ The Electron Microscope Resolves the Fine
Structure of the Cell.▪ Images of Surfaces Can Be Obtained by Scanning
Electron Microscopy▪ Metal Shadowing Allows Surface Features to Be
Examined at High Resolution by Transmission Electron Microscopy
▪ Freeze-Fracture and Freeze-Etch Electron Mi-croscopy Provide Views of Surfaces Inside the Cell
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Resolution, resolution!!!
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Electron Microscopy
Why electron beams? The wavelength, much shorter than of light
Types Transmission electron microscope (TEM) Scanning electron microscope (SEM) - sur-
face contoursTEM SEM
R = 0.61λ/ N.A.
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Electromagnetic lenses
• a coil of wire through which current flows• magnetic field in the center.• electrons traveling in their path down the central
hole.
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TEM
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Dark (Electron dense)
(Electron transparent)
Bright (Electron lucent)
Contrast in TEM
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• Bright (Electron lucent) - electrons readily passed
• Dark (Electron dense) - electrons absorbed
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Transmission electron microscope (TEM)
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SEM TEMDIC
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SEM
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Scanning electron microscope (SEM)
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Metal Shadow-ing
• high resolution three-dimen-sional replica of the specimen
• heavy metal shadowing of bacteria to demonstrate flagella
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Freezing Techniques
•Freeze-Fracture •Freeze-Etch
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TEM Section and Freeze Fracture