Introduction to optical microscopy

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PREFACE TO FIRST EDITION
For anyone who has taught a graduate or upper-level undergraduate course in optical
microscopy, the problem often arises of which textbook to use. While there are many excellent
textbooks that deal with various applications of optical microscopy, and still more that deal
with optics in general, very few textbooks deal specifically with optical microscopy from a
physics or engineering standpoint. A notable exception is, or rather was, Pluta’s Advanced
Light Microscopy series, but unfortunately this has become out of date and is currently out of
print. My own solution to the problem has been to heavily rely on course notes; however, over
the years, these have become steadily more scattered to the point of becoming unusable. And
so this book was conceived largely out of necessity, my goal being to provide a self-contained
overview of the foundations and techniques of optical microscopy suitable for teaching at the
graduate or advanced undergraduate level.
This book can be divided into two parts. The first part (Chapters 1–8) deals with properties
of light, light propagation, and light detection and is meant to provide the groundwork for
the remainder of the book. Most of this first part is centered on Fourier optics and statistical
optics and is intended as an overview of some of the fundamental concepts in these fields,
ranging from imaging transfer functions to partial coherence theory. Much of this material can
be found in, and indeed was drawn from, many excellent textbooks, in particular, Goodman’s
Introduction to Fourier Optics and Statistical Optics. I tailored this material as much as possible
to optical microscopy and included chapters on 3D imaging and radiometry for completeness.
The first of these chapters provides a synthesis of the generalized imaging functions involved
in 3D microscopy. The second discusses concepts of étendue and throughput as applied to
microscope layouts. These concepts seem to have fallen by the wayside in many optics
textbooks, perhaps because of their limited connection to Fourier optics. Nevertheless, they
remain highly useful for facilitating a “first pass” intuitive understanding of optical layouts.
The second part of this book (Chapters 9–18) is specifically dedicated to microscopy
techniques, in particular, to contrast mechanisms and to the repertoire of tricks, old and new,
that has been devised for revealing contrast. I made no attempt to present these techniques in
chronological order or in their historical context. Instead, I chose to group them under common
themes to emphasize their underlying relations to one another. Indeed, techniques that might
have appeared dissimilar when treated separately turn out in many cases to be implementations
of the same basic idea. The material in these chapters was more difficult to distill from the
literature and, for this reason, is likely to constitute the main interest of this book.
Chapters 9–12 deal with nonfluorescence imaging techniques, particularly those that revolve
around phase contrast and interferometry. Chapter 9 is an introduction to the physical mech-

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