BIO 595R-595W/BMS 517C |
TRANSMISSION ELECTRON MICROSCOPY |
SPRING 2004 |
Class Notes
I. THE MICROSCOPE pgs. 1-114
A. PRINCIPLES OF THE TRANSMISSION ELECTRON
MICROSCOPE (TEM) pgs. 1-28
1. Origin of the Electron Microscope pg. 1
2. Comparison of Light and Electron Microscopes pgs. 1-4
3. Photons/Electrons pgs. 5-11
4. Optics pgs. 12-19
5. Electron Beams / Electron Optics / Magnetic and
Electrostatic Lenses pgs. 19-28
B. DESIGN OF THE TEM pgs. 29-57
1. Electron gun pgs. 29-32
2. Condenser Lens pgs. 32-35
3. Lens Aberrations pgs. 35-42
4. Objective Lens and Specimen Stage pgs. 42-48
5. Projector Lens pgs. 48-50
6. Camera and Viewing System pgs. 51-52
7. Vacuum System pgs. 52-56
8. Electrical System pgs. 56-57
C. CONTRAST AND IMAGE FORMATION pgs.
58-64
1. Electron Scattering pgs. 58-60
2. Amplitude/Phase Contrast pgs. 60-63
3. Phase Contrast Transfer Function pg. 63
4. Multiple Scattering in Thick Specimens pg. 64
5. Other Methods for Enhancing Contrast pg. 64
D. ALIGNMENT/ADJUSTMENT OF THE TEM pgs.
65-73
1. Introduction pg. 65
2. Alignment of the Microscope pgs. 65-70
3. Disturbances to Microscope Performance pgs. 70-73
E. OPERATION OF THE TEM pgs. 74-93
1. Choice of Voltage pgs. 74-75
2. Choice of Apertures pg. 75
3. Specimen Stage/Holder pg. 75
4. Choice of Magnification pg. 76
5. Focusing pgs. 77-80
6. Magnification Calibration pgs. 80-82
7. Resolution Tests pgs. 82-84
8. Image Intensifier / TV Display pgs. 84-85
9. Microscope Maintenance pgs. 85-86
10. Photography pgs. 86-93
F. OTHER MODES OF TEM pgs. 94-114
1. Electron Diffraction pgs. 94-100
2. Dark Field pgs. 101-106
3. High Resolution pgs. 106-107
4. Tilting and Stereo Microscopy pgs. 107-112
5. Low Temperature pg. 112
6. Energy Loss pgs. 112-113
7. X-ray Microanalysis pgs. 113-114
Top of Notes
II. THE SPECIMEN pgs. 115-174
A. BIOLOGICAL SPECIMEN PREPARATION TECHNIQUES pgs. 115-161
1. Specimen Support Films for TEM
pgs. 115-121
2. Thin Sectioning (Fixation/Dehydration/Embedding/Staining)
pgs. 121-135
3. Negative Staining
pgs. 136-142
4. Metal Shadowing
pgs. 143-149
5. Unstained Specimens
pgs. 149-155
6. Freeze Drying/Etching/Fracture
pgs. 155-158
7. Autoradiography
pgs. 158-161
B. RADIATION EFFECTS pgs. 162-174
1. Introduction
pg. 162
2. Dose/Dose Rate
pg. 162
3. Primary Effects of Radiation Damage to Biological Specimens
pgs. 162-164
4. Secondary Effects of Radiation Damage to Biological Specimens
pgs. 165-167
5. Ways to Measure Damage/Critical Dose
pgs. 167-169
6. Procedures to Reduce Radiation Damage
pgs. 170-172
7. Relation Between Contrast, Resolution and Radiation Damage
pg. 172
8. Radiation Effects in Negatively-Stained Specimens
pgs. 172-174
9. Radiation Effects in Frozen-hydrated Specimens
pg. 174
Top of Notes
III. THE MICROGRAPH pgs. 175-253 (Some parts of this section
are not currently available)
A. INTRODUCTION TO IMAGE ANALYSIS OF BIOLOGICAL SPECIMENS pgs. 175-176
1. Image Analysis Techniques: Real and Reciprocal Space
Techniques
pg. 175
2. Fourier Image Processing
pgs. 175-176
3. Periodic/Non-Periodic Specimens
pg. 176
4. Applications and Advantages of Image Processing
pg. 176
B. SOURCES OF NOISE IN TEM IMAGES OF BIOLOGICAL SPECIMENS pg. 177
1. Specimen Support Film
pg. 177
2. Specimen pg. 177
3. Microscope pg. 177
4. Photography pg. 177
C. CRYSTALS, SYMMETRY AND DIFFRACTION pgs. 178-216
1. Definition of Terms
pgs. 178-179
2. Crystals
pg. 179
3. Lattices pgs. 179-181
4. Crystal Structure pgs. 181-182
5. Symmetry pgs. 183-195
6. Diffraction pgs. 195-216
D. FOURIER IMAGE PROCESSING TECHNIQUES pgs. 217-253
1. Optical Diffraction ...
2. Optical Filtering ...
3. Digital Fourier Analysis of Electron Micrographs (4.6MB) pgs. 217-240
4. 2D and 1D Digital Fourier Reconstruction Methods (4.6MB)
pgs. 241-253
Power Point Lecture (2.8MB)
E. ANALYZING IMAGES OF SPECIMENS OF DIFFERENT TYPES
1. Planar Specimens
2. Particles with Rotational Symmetry
3. Helical
4. Three-dimensional Crystals
5. Asymmetric Particles
F. SPECIMEN PREPARATION AND MICROSCOPY
1. Sample Preparation
2. Imaging Conditions
G. REAL SPACE AND OTHER RECONSTRUCTION METHODS
H. INTERPRETATION AND DISPLAY OF IMAGE ANALYSIS RESULTS
Top of Notes
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