Light Microscopic Techniques in Biology and Medicine

ISBN-13: 9789401014168 / Angielski / Miękka / 2011 / 336 str.

J. James

Opis

Up to about twenty-five years ago, virtually the entire field of microscopy could be overseen and even practized by any active research worker. The rapid evolution which microscopy in its broadest sense has since undergone and which has contributed greatly to our insight in many fields of biological science and medicine has, however, lead to a progressive specialisation. Both experienced investigators in clinical and biological laboratories and post- graduate students, confronted with a limited number of microscopic tech- niques in their daily research work, have increasing difficulty in keeping (or obtaining) a general idea of the many time-honoured and new possibilities which microscopy has to offer. This book has been written with the aim of presenting general informa- tion on light microscopic techniques, at a level somewhere in between booklets like those provided by microscope manufacturers (which are often too much focussed on the production program of a particular make) and very advanced treatises with a thorough mathematical treatment of all phenomena concerned. The physically oriented texts moreover often do not sufficiently take into account the practical situation in a medical or biolog- ical laboratory; on the other hand, the value of really understanding what one is doing in using a microscopic technique is often underestimated. At- tempt has been made, therefore, to present sufficient background informa- tion necessary for a rational application of the different microscopical tech- niques in their mutual relationship.

Spis treści

I: Conventional techniques of microscopic observation.- 1 Some Essentials of Geometrical Optics.- Geometrical optics and wave optics.- Resolving power and the eye.- The simple microscope.- Lens aberrations.- Properties of lens combinations.- The compound microscope.- Suggestions for further reading.- 2 From Optical Principle to a Practical Instrument.- Some notes about the history of the microscope.- The stand and its parts.- Stereoscopic vision and the microscope.- Literature cited and suggestions for further reading.- 3 Objectives.- Numerical aperture.- Immersion-objectives.- The cover glass effect.- Types of objective.- Qualifications and performance of an objective.- Special objectives.- — mirror- or reflecting objectives.- — objectives for use with invisible light.- — photomicrographic objectives.- Literature cited and suggestions for further reading.- 4 Eyepieces or Oculars.- Main types of eyepiece.- Exit pupil and the eye.- Eyepiece and field of view.- The meaning of eyepiece magnification.- Special types of eyepiece.- Literature cited and suggestions for further reading.- 5 Relation of Objective and Eyepiece.- Resolving power.- Objective, eyepiece and the eye.- Depth of field.- Literature cited and suggestions for further reading.- 6 Condenser and Illumination.- The function of the condenser.- Critical illumination and Köhler illumination.- Types of condenser.- Special types of illumination; incident illumination.- The light source.- Literature cited and suggestions for further reading.- 7 Specimen, Microscope and Observer; Microscopy in Practice.- The microscopic object as an optical model.- The technique of microscopic observation.- — Position of the observer and the placing of the microscope.- — General rules for setting up a microscope.- — Use of immersion objectives.- — Light and illumination in practice.- — On the way through the object.- — Microscopy for observers wearing spectacles.- Maintenance and small technical difficulties.- — Care of the stand and the cleaning of optical components.- — Frequently occurring minor technical troubles.- Literature cited and suggestions for further reading.- II: Advanced techniques of microscopy.- 8 Special Techniques of Illumination.- Oblique illumination.- Dark-field illumination.- Fluorescence microscopy.- — General principles.- — Optical arrangements with a fluorescence microscope.- Literature cited and suggestions for further reading.- 9 Special Techniques of Image Formation.- Phase contrast microscopy.- — Basic principles.- — Practical realization of the phase contrast principle.- — Some further details about the phase contrast image with different objects.- Interference microscopy.- — Basic principles; the meaning of a refractive index.- — Interferometric measuring systems.- — Differential interference contrast.- Polarization microscopy.- — Basic principles of birefringence.- — The polarization microscope.- — Some applications of polarization microscopy in biological research.- Literature cited and suggestions for further reading.- 10 Recording and Reproduction of Microscopic Images.- Photomicrography.- — General principles.- — Photomicrographic equipment.- — The photomicrographic exposure.- — Contrasts in the negative.- — Colour photomicrography.- Some special techniques in photomicrography.- — Microflash.- — Stereophotomicrography.- — Holography.- — Cinemicrography.- Other techniques for registration and reproduction of microscopic images.- — Drawing devices.- — Microprojection.- — Television-microscopy.- Literature cited and suggestions for further reading.- 11 Measurements with the Microscope.- General introduction.- Morphometric analysis.- — Measurement of length in a focussing plane.- — Measurement of distances along the optical axis.- — Measurement of length oblique to the optical axis.- Measurement of areas and volumes.- — Measurement of areas.- — Measurement of volumes; stereology.- Automatic and semi-automatic image analysis.- Microspectrophotometry and microphotometry.- — Microspectrophotometry.- — Microphotometry.- Microspectrofluorometry and microfluorometry.- Literature cited and suggestions for further reading.- 12 Microscopy with Invisible Electromagnetic Radiation.- Microscopy and the electromagnetic spectrum.- Ultraviolet microscopy.- Infrared microscopy.- Use of X-rays.- — Some fundamental properties of X-rays and their applications.- — X-ray microscopy.- — Contact microradiography.- — Projection-microradiography.- Literature cited and suggestions for further reading.- Appendix I Table of refractive indices.- Appendix II Four-lingual vocabulary of some commonly used technical terms in microscopy.