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Introduction to Microscopy by Means of Light, Electrons, X Rays, or Acoustics
Name: Introduction to Microscopy by Means of Light, Electrons, X Rays, or Acoustics
Author: theodore george rochow
Pages: 462
Year: 1994
Language: English
File Size: 20.37 MB
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L1brarv of Congress Catalog1ng 1n Publ1cat1on Data Rochow, Theodore George. Introduct1on ta m1croscopy by means of 11ght, electrons, X rays, ar acoust1cs 1 Theodore George Rochow and Paul Arthur Tucker. 2nd ed. p. cm. Rev. ed. of: An 1ntroduct1on ta m1croscopy by means of 11ght, electrons, X rays, ar ultrasound. c1978. Includes b1b11ograph1cal references and 1ndex. ISBN 978 1 4899 1515 3 ISBN 978 1 4899 1513 9 (eBook) DOI 10.1007/978 1 4899 1513 9 1. M1croscopy. I. Tucker, Paul Arthur. II. Rochow, Theodore George. Introduct1on to m1croscopy by means of 11ght, electrons, X rays, ar ultrasound. III. T1tle. OH205.2.R63 1994 502 . 8 2 dc20 DNLM/DLC for L1brary of Congress ISBN 978 1 4899 1515 3 1994, 1978 Springer Science+Business Media New York Originally published by Plenurn Press, New York in 1994 Softcover reprint ofthe hardcover 2nd edition 1994 All rights reserved 94 15345 CIP No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher


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Preface Following three printings of the First Edition (1978), the publisher has asked for a Second Edition to bring the contents up to date. In doing so the authors aim to show how the newer microscopies are related to the older types with respect to theoretical resolving power (what you pay for) and resolution (what you get). The book is an introduction to students, technicians, technologists, and scientists in biology, medicine, science, and engineering. It should be useful in academic and industrial research, consulting, and forensics; how ever, the book is not intended to be encyclopedic. The authors are greatly indebted to the College of Textiles of North Carolina State University at Raleigh for support from the administration there for typing, word processing, stationery, mailing, drafting diagrams, and general assistance. We personally thank Joann Fish for word process ing, Teresa M. Langley and Grace Parnell for typing services, Mark Bowen for drawing graphs and diagrams, Chuck Gardner for photographic ser vices, Deepak Bhattavahalli for his work with the proofs, and all the other people who have given us their assistance. The authors wish to acknowledge the many valuable suggestions given by Eugene G. Rochow and the significant editorial contributions made by Elizabeth Cook Rochow. Raleigh, North Carolina Theodore G. Rochow Paul A. Tucker vii


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Contents 1. A Brief History of Microscopy 1.1. Introduction............................................ 1 1.2. Correcting for Aberrations............................... 6 1.3. Condensers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 1.4. Dark Field Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 1.5. Polarized Light Microscopy.............................. 9 1.6. Near Field Scanning Light Microscopes . . . . . . . . . . . . . . . . . . . 11 1.7. Light Microscope Manufacturers.......................... 11 1.8. Transmission Electron Microscopes....................... 12 1.9. Scanning Electron Microscopes........................... 15 1.10. Electron Probe Microanalyzers . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 1.11. Field Emission Microscopes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 1.12. Scanning Tunneling Microscopes . . . . . . . . . . . . . . . . . . . . . . . . . 16 1.13. Scanning Acoustic Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 1.14. Atomic Force Microscopes............................... 18 1.15. X Ray Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 1.16. X Ray Laser Microscopes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1.17. Microscopy Society of America........................... 19 1.18. Summary.............................................. 20 2. Definitions, Attributes of Visibility, and General Principles 2.1. Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 2.2. Attributes of Visibility.. . .. . .. .. .. . .. .. . .. . .. .. . . . .. . .. . . 25 2.2.1. Correcting for Aberrations........................ 25 2.2.2. Sample Quantity and Quality . . . . . . . . . . . . . . . . . . . . . 25 2.2.3. Focus Depth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 2.2.4. Focus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 2.2.5. Illumination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 2.2.6. Radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 ix


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X Contents 2.2.7. Anisotropy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 2.2.8. Magnification.................................... 31 2.2.9. Stereoscopy..................................... 32 2.3. General Principles. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 2.3.1. Specimen Structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 2.3.2. Specimen Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 2.3.3. Specimen Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 2.3.4. Experimentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 2.3.5. Specimen Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 2.3.6. Specimen Behavior. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 2.3.7. Photomicrography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 2.3.8. Video. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 2.4. Summary............................................... 35 3. Simple and Compound Light Microscopes 3.1. Limits of Resolution by the Eye........................... 37 3.2. Simple Microscopes: One Lens Systems.................... 38 3.3. Compound Microscopes: Two or More Lens Systems . . . . . . . 40 3.4. Stereocompound Microscopes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 3.4.1. Illuminating with Stereomicroscopes............... 48 3.4.2. Preparing the Specimen . . . . . . . . . . . . . . . . . . . . . . . . . . 49 3.5. Biological Microscopes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 3.5.1. Objectives....................................... 50 3.5.2. Eyepieces....................................... 51 3.5.3. Condensers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 3.5.4. Illumination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 3.5.5. Ultraviolet and Infrared Light . . . . . . . . . . . . . . . . . . . . . 58 3.5.6. Anisotropy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 3.5.7. Magnification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 3.5.8. Photomicrography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 3.6. Summary............................................... 59 4. Compound Microscopes Using Reflected Light 4.1. Studying Surfaces by Reflected Light...................... 61 4.2. Resolving Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 4.3. Contrast. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 4.4. Correcting for Aberrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 4.5. Specimen Cleanliness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 4.6. Focus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 4.7. Illumination............................................ 68 4.8. Radiation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 4.9. Magnification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 4.10. Field of View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77


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Contents 4.11 4.12. 4.13. 4.14. 4.15. 4.16. 4.17. 4.18. 4.19. 4.20. 4.21. Glare ................................................. . Depth ................................................ . Working Distance ...................................... . Structure .............................................. . Morphology ........................................... . Information about the Specimen ......................... . Experimentation ....................................... . Specimen Behavior ..................................... . Specimen Preparation .................................. . Photomicrographic Techniques .......................... . Summary ............................................. . xi 78 79 80 81 82 83 83 83 86 87 87 5. Microscopy with Polarized Light 5.1. Overhead Projections.................................... 89 5.2. Anisotropy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 5.3. Numerical Aperture and Interference Figures . . . . . . . . . . . . . . 94 5.4. Resolution: Specimen Interaction and Polarized Light . . . . . . . 95 5.5. Contrast: Michel Levy Interference Chart. . . . . . . . . . . . . . . . . . 97 5.5.1. Personal Interpretation of Interference Colors....... 99 5.5.2. Retardation Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 5.5.3. Specimen Thickness. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 5.6. Correcting for Aberrations Due to Strain . . . . . . . . . . . . . . . . . . 101 5.7. Cleanliness: Freedom from Interference Films.............. 102 5.8. Focus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 5.9. Illumination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 5.10. Radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104 5.11. Magnification.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104 5.12. Field of View of an Interference Figure.................... 105 5.13. Glare . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105 5.14. Depth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 5.15. Working Distance....................................... 106 5.16. Specimen Structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 5.17. Specimen Morphology................................... 107 5.18. Information about the Specimen . . . . . . . . . . . . . . . . . . . . . . . . . . 107 5.19. Experimentation........................................ 107 5.20. Specimen Behavior. . . . . . . . . . . . . . .. . . . . . .. . . . . . . . . . . .. . . . 108 5.21. Specimen Preparation................................... 108 5.22. Photography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108 5.23. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109 6. Microscopical Properties of Fibers 6.1. Introduction............................................ 113 6.2. Fiber Morphology.. .. . .. .. .. . .. .. .. . .. . .. .. .. .. .. . .. .. .. 113


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