Light and Electron MicroscopyOptical and electron microscopes are often used effectively despite little knowledge of the relevant theory or even of how a particular type of microscope functions. Eventually however proper use interpretation of images and choices of specific applications demand an understanding of fundamental principles. This book describes the principles of operation of each type of microscope currently available and of use to biomedical and materials scientists explains the mechanisms of image formation (contrast and its enhancement) accounts for ultimate limits on the size of observable details (resolving power and resolution) and finally provides an account of Fourier optical theory. Principles behind the photographic methods used in microscopy are described and there is some discussion of image processing methods. Throughout the text emphasises the underlying similarity of all microscope systems and recognising that biologists may often be uncomfortable with mathematical approaches every effort has been made to present concepts verbally. Where mathematical treatment is indispensible the nature of its contribution is made explicit. |
Contents
Introduction | 1 |
definitions and brief history | 2 |
13 Microscope design | 4 |
14 Mathematical aspects | 6 |
Light and electrons | 7 |
22 Geometrical optics | 10 |
23 Wave optics physical optics | 12 |
24 Characteristics of wave motions | 15 |
111 Phasecontrast principles | 150 |
112 The phasecontrast microscope | 152 |
113 The differentialinterferencecontrast DIC microscope | 154 |
114 The modulationcontrast microscope MCM | 158 |
115 Reflectioninterference microscopy | 160 |
116 Interferometer microscopes | 161 |
117 Quantitative intracellularmass measurements by interferometric microscopy | 166 |
Polarizing microscopy | 168 |
25 Limits of lightelectron parallelism | 21 |
Wave interactions | 23 |
32 Huygens principle | 24 |
33 Resultant vibrations | 25 |
34 Superposition of waves of different frequencies Fourier optics | 29 |
35 Applications of Fourier theory to basic properties of radiation | 36 |
Interference effects and diffraction patterns | 39 |
42 Interference and diffraction | 40 |
44 Diffraction patterns | 43 |
45 Holography | 49 |
Polarized light | 51 |
52 Circularly and elliptically polarized light | 52 |
53 Interactions of polarized light with oriented matter | 56 |
54 Sources of planepolarized light | 61 |
55 Polarizers analyzers and compensators | 63 |
Lenses | 65 |
62 Focusing properties of curved surfaces | 66 |
63 Focal points focal lengths and focal planes | 68 |
65 Magnification | 71 |
67 Image location | 73 |
69 Lens errors | 79 |
Imaging microscopy and diffraction | 88 |
73 Transforms and inverse transforms the optical diffractometer | 91 |
74 Diffraction and reciprocal space | 92 |
the phase problem | 93 |
Contrast | 95 |
82 Lightoptical contrast | 97 |
83 Electronoptical contrast | 100 |
84 Contrast transfer functions | 112 |
85 Massthickness contrast | 116 |
Resolution | 118 |
92 Fourier optics and limiting resolution | 119 |
93 Coherence properties and resolution | 121 |
95 Numerical aperture and the immersion principle | 126 |
96 Extension of the classical resolution limit | 128 |
97 Conflicts between contrast and resolution | 129 |
The light microscope | 131 |
102 Magnification and calibration | 139 |
103 Depths of field and focus for light microscopes | 140 |
104 Alternative modes of optical microscopy | 142 |
Imaging of phase objects | 149 |
122 The polarizing microscope | 169 |
123 Basic concepts of polarizing microscopy | 170 |
124 Compensators | 173 |
125 Biological polarizing microscopes | 177 |
126 Measurements with the polarizing microscope | 179 |
Prospects for biological xray microscopy | 184 |
132 Xray sources | 186 |
133 Types of xray microscopes | 187 |
The conventional transmission electron microscope | 192 |
142 Alternative operating modes | 206 |
Scanning microscopes | 212 |
152 Types of electron scanning microscopes | 213 |
153 The SEM | 214 |
154 The STEM | 221 |
Practical aspects of electron microscopy | 228 |
162 Depths of Field and focus and threedimensional structure | 235 |
163 Determination of molecular weight | 236 |
164 Operation of the electron microscope | 238 |
The quest for ultimate electron microscopic resolution | 247 |
172 Electron imaging | 250 |
173 Image processing using Fourier analysis | 253 |
174 The darkfield image | 260 |
175 Ultimate resolution | 264 |
Innovations in microscope development | 270 |
183 The neutron microscope | 271 |
186 Nuclearmagneticresonance microscopy | 275 |
187 The acoustic microscope | 276 |
188 Superresolving instruments and confocal systems | 278 |
189 Videoenhanced light microscope systems | 284 |
Photography | 286 |
192 Formation of the latent image | 287 |
193 Chemical processing | 288 |
194 Properties of photographic emulsions | 291 |
195 Basic features of color photography | 296 |
Image location | 298 |
B Sign conventions | 300 |
F Raytracing equations for exact image location | 301 |
303 | |
307 | |
Other editions - View all
Common terms and phrases
Airy disc amplitude aperture angle atoms azimuth beam damage birefringence brightfield chromatic aberration component condenser contrast corresponding darkfield density described detector diffraction pattern direction dmin E-ray effects electromagnetic electron beam electron microscopy elements emission emulsion energy loss equation film focal length focal plane focus focused Fourier optics Fourier transform high-resolution illumination image plane incident inelastic instruments intensity interactions interference John Wiley lens light microscopy limited magnetic magnification Methods in Biology microscope object objective lens observed obtained optic axis Optical Methods particles permission perpendicular phase-contrast photographic plane-polarized plate polarized position produce radiation rays reflection refractive index resolving power scanning scattering sect shown in Figure simple harmonic motion Slayter slit space spatial frequencies specific specimen spherical aberration structure surface thickness tion transfer function transmission transmitted values velocity vibration wave front wavelength Wiley & Sons Wollaston prism x-ray