Contrast techniques in light microscopy

Cover......Page 1
Half Title......Page 2
Title Page......Page 6
Copyright Page......Page 7
Table of Contents......Page 8
Preface......Page 12
Safety......Page 13
 Why use a microscope?......Page 14
 The need for contrast enhancement......Page 15
 Assessment of the image......Page 17
 References......Page 19
 Light and the specimen......Page 20
 Absorption, transmission and reflection of light by the specimen......Page 21
 Scattering and diffraction......Page 25
 Refraction and polarized light......Page 26
 Phase change......Page 29
 Fluorescence......Page 30
 Stray light......Page 31
 References......Page 32
3. Bright-field and Dark-ground Techniques......Page 34
 Manipulation of the illumination......Page 35
  Oblique light......Page 37
 Enhancing contrast by use of colour–natural colour, staining and illumination changes......Page 39
 Colour filters......Page 42
 Dark-ground imaging for transmitted light......Page 43
 Dark-ground epi-illumination......Page 48
 Rheinberg illumination......Page 49
 Stray light......Page 53
 References......Page 54
4. Refractive Differences between Specimen and Mountant; the ‘Becke Line’......Page 56
 Dispersion staining......Page 57
 References......Page 61
5. Polarized-light Techniques......Page 62
 The polarized-light microscope......Page 63
 Observations with a single polar......Page 65
 Observations between crossed polars; first-order red plate......Page 67
 The use of polarized light in reflected-light microscopy......Page 70
 References......Page 71
6. Phase Contrast and Modulation Contrast......Page 72
 Theory of phase contrast......Page 73
 The construction of the phase-contrast microscope......Page 75
 Variants of phase contrast......Page 79
 The phase-contrast image......Page 81
 Phase contrast in reflected light......Page 82
 Construction of the modulation-contrast microscope......Page 83
 Setting-up phase contrast......Page 86
 References......Page 88
7. Interference Contrast......Page 90
 Differential interference contrast......Page 93
 The DIC microscope......Page 94
 The transmitted-light DIC image......Page 99
 Setting-up DIC for transmitted light......Page 100
 Setting-up DIC for reflected light......Page 101
 References......Page 102
8. Fluorescence Microscopy......Page 104
 Types of fluorescence......Page 105
 The fluorescence microscope......Page 107
 Light sources for fluorescence......Page 108
 Objectives and filters......Page 109
 References......Page 113
 Photographic techniques of contrast enhancement......Page 114
 Film choice, negative development and printing......Page 115
 Image enhancement by video techniques......Page 117
 Image modification by digital techniques......Page 118
 Contrast enhancement by manipulation of the grey-level histogram......Page 120
 References......Page 122
 A vector theory approach to phase contrast......Page 124
 Reference......Page 127
Index......Page 128