Table Of ContentFrontispiece
Top: An autoradiogram showing the deposition of
iodine-131 in the kidney of a rat. Iodine-labeled pro
teins were injected intravenously into a rat which was
sacrificed a few hours later. The blackened areas show
the deposition of the iodine, and presumably the pro
teins, in the glomeruli. The wet-mounting technique
with an NTB plate was used with metanil yellow stain
ing after photographic processing.
Courtesy P. J. Fitzgerald (unpublished), Sloan-Ket-
tering Institute.
Bottom : An autoradiogram showing the deposition of
sulfur-35 in the skin of a newborn mouse. One mc of
inorganic sulfur-35 was injected intraperitoneally into
the mother 24 hours before birth. The autoradio-
graphie evidence is the random distribution of grains
over three cell nuclei. These are probably mast cells.
The developing hair follicles show no uptake. The
permeable-base stripping film technique was used with
haematoxylin and celestin blue staining after photo
graphic processing.
Courtesy A. Glücksmann, Alma Howard, and S. R.
Pelc, Radiotherapeutic Research Unit, Hammersmith-
Hospital, London.
AUTORADIOGRAPHY
IN BIOLOGY
AND MEDICINE
GEORGE A. BOYD
Director, Arizona Research Laboratories, Phoenix, Ari
zona. Formerly, Professor of Biophysics, University of
Tennessee, and Senior Scientist, Oak Ridge Institute of
Nuclear Studies
ACADEMIC PRESS INC., PUBLISHERS
NEW YORK, 1955
COPYRIGHT 1955, by
ACADEMIC PRESS INC.
125 EAST 23RD STREET
NEW YORK 10, N. Y.
NO PART OF THIS BOOK MAY BE REPRODUCED IN
ANY FORM, BY PHOTOSTAT, MICROFILM, OR ANY
OTHER MEANS, WITHOUT WRITTEN PERMISSION
FROM THE PUBLISHERS.
Library of Congress Catalog Card Numker: 54-11051
PRINTED IN THE UNITED STATES OF AMERICA
DEDICATED
TO
DR. W. W. STEFFEY
Preface
Autoradiography in biology and medicine is a marriage of the arts of
histology and photography. As in the fine arts, there are as many ways
of doing autoradiography as there are doers. There are, however, scien
tific pathways to follow in learning the art.
This book is divided into three sections. In the first section is given
much of the background for the scientific approach to the art given in
the second section. The many techniques given in the second section are
in "cook-book" style and can be followed explicitly by the laboratory
technician.
There is no guarantee that a technique will work in every laboratory,
in every part of the country, or for all materials to be used. It is almost
impossible for the originator of a technique to recognize some essential
steps and transfer them to me by conversation and I to the reader
through the printed page.
I have attempted to elicit, from the authors, all the little details
which we so often unconsciously perform in the laboratory; those details
which we sometimes have sound, but intuitive, reasons for doing yet
forget to include in a published paper. Sometimes these unconscious acts
are mere habit. They may make for success of a technique. This is the
personal art : the reason some are successful where others are not.
I have written this book for the researcher who has never done auto
radiography. The technique to use will be indicated by the nature of his
investigation. A study of the first section will help him decide on the
materials to be used in the experiment to obtain meaningful information.
It will be obvious that many of the steps of a technique can be varied.
None of them are sacred. Rather, they are diving boards from which
you can take a plunge into the pool of variables and come up with your
own pet combination—a new technique.
The quantitative aspects of autoradiography have not been included.
The book was growing to unmanageable proportions. Also, much good
vii
viii PREFACE
work in this field is in process and has not been published. A section on
quantitative autoradiography at this time would be premature.
The third section is an extensive bibliography handled in, I believe, a
novel manner. This is discussed in detail in the Introduction to the
Bibliography.
I owe my interest in autoradiography to Professor William Bale of the
University of Rochester who suggested I investigate its potentialities in
the study of polonium distribution in tissue sections. I am grateful to
him, even though the study of the technique temporarily diverted me
from my earlier interest of radiocolloids in tissues.
The decision to write a book came through a suggestion of Dr. Kurt
Salomon, also of the University of Rochester. I thank him for this, and
am sure I shall feel more kindly toward him after corrections are made
and the presses start.
GEORGE A. BOYD
Acknowledgments
I wish especially to thank Dr. Harry Blair and Dr. J. Newell Stannard
of the University of Rochester for their interest in the work. It is a
pleasure to acknowledge the interest shown in the writing by Dr. C. L.
Cornar, Dr. R. T. Overman, and Dr. M. Brucer. I am especially grateful
to Dr. Overman for his encouragement and kindly criticism during the
preparation of the manuscript.
I owe a special debt of gratitude to Dr. D. R. Smith who helped me
set up a punch card system, without which I could not have written the
book. Assistance in searching was generously given by Mr. R. A. Schlueter,
Chief Librarian of ORINS. The Misses Marion Garber and Louise
Markel and my secretary, Mrs. Richard Broad, did the painstaking job
of checking the literature references.
Over a period of several years, various members of the Eastman
Kodak Company have been most generous with their time in discussing
several phases of this book, reading some of the chapters, and advising
on the presentation of material. It should not be inferred that they neces
sarily put their stamp of approval on my statements. Any errors of photo
graphic theory and facts are mine in spite of their excellent teaching.
For making this a better book, I want to thank Drs. Cyril Staud,
John Spence, Julian Webb, Herman E. Seeman, T. H. James, and B. H.
Carroll, all of the Eastman Kodak Research Laboratories, Mr. William
Swann and Mr. R. Tobey of the Sales Department, and Mr. William
Cornwell, Editor of Medical Radiography and Photography.
I am indebted to Mr. John Barione of Saint Norbert College who
read almost the entire manuscript. He looked at it through the eyes of an
histologist, and much improved the presentation.
The publishers have been most considerate and helpful.
And last I wish to thank the one in the background who furnished
the inspiration and the understanding of my moods of elation when the
writing went well and of depression when it went sour, who deflected
the romping and yelling children when I wished to write, and who was
my chief critic : Carolyn, my wife.
îx
CHAPTER I
Introduction
1. HISTORICAL
On Monday, January 20, 1896, the French Academy of Sciences
met in Paris as usual for the reading of papers and the discussion of
the latest scientific events. Henri Becquerel attended as was his custom
and as had been the custom of his father and grandfather. All three
were physicists and had held the research post of Professor of Physics
at the Museum of Natural History. Henri also held the teaching post at
the Ecole Polytechnique.
His father, Edmond, and Henri had studied fluorescence. With this
background Henri attended the meeting where he was to hear exciting
news. Henri Poincaré showed the X-ray pictures sent by Rontgen from
Vienna and discussed the paper which Rontgen had sent to press
December 29, 1895. Poincaré suggested that the X-rays might be a
portion of the fluorescence radiation which Rontgen had seen on the
wall of his glass X-ray tube.
This set Becquerel thinking. The next day he started his investiga
tions of several fluorescing materials but most carefully of a flat crystal
of potassium uranyl sulfate which he had made. He wrapped a photo
graphic plate in black paper, placed a crystal on top, and exposed the
combination to sunlight. On developing the plate after several hours
exposure, he was pleased to see a faint outline of the crystal—the first
autoradiogram, although unrecognized. Becquerel thought the X-ray
like radiation was associated with the fluorescence. He continued his
experiments and showed that the blackening was always associated
with, and only with, salts or solutions containing uranium, even though
fluorescence was not always present. In the same year he showed that
pure uranium metal produced the blackening. Nevertheless, he con-
3
4 THEORY
tinued to think that fluorescence, even though not always seen, was in
some way connected with the phenomenon.
Thus, through serendipity, the first radioactivity autoradiogram was
made but not recognized. Becquerel's inability to give up the idea of
fluorescence prevented him from suspecting an entirely new phenom
enon. Although he discovered the first raw facts of radioactivity, he
failed to understand and interpret them. It remained for Marie Curie,
in the Spring of 1898, to establish by ionization measurements the rela
tion of radiation quantity with the quantity of the uranium element.
Thus, she was the first to recognize radioactivity.1
It seems apropos to digress for one paragraph to consider the nature
of discovery, since autoradiography can easily produce raw facts which,
in themselves, do not constitute discovery. I shall let Claude Bernard
comment :2
"We usually give the name of discovery to recognition of
a new fact : but I think that the idea connected with the dis
covered fact is what really constitutes the discovery. Facts
are neither great nor small in themselves. A great discovery
is a fact whose appearance in science gives rise to ideas
shedding a bright light which dispels many obscurities and
shows us new paths. There are other facts which, although
new, teach us but little; they are therefore small discoveries.
Finally, there are new facts which, athough well observed,
mean nothing to anyone; they remain, for the time being,
detached and sterile in science; they are what we may call
crude or raw facts."
Autoradiography was first intentionally used for locating radio
activity in minerals (12). The first published ( 1904 ) biological appli
cation was that of E. S. London of the Imperial Institute of Experi-
11 am indebted to Professor Alfred Römer of St. Lawrence University,
Canton, New York, for permission to see the manuscript of Chapter 2 of his
forthcoming book, "The Development of Atomic Physics," from which I have
drawn heavily for the facts of the first four paragraphs. For a more detailed
and excellently documented account of Becquerel's investigation, see Professor
Romer's book.
2J. M. D. Olmsted and E. H. Olmsted, Claude Bernard and the Experi
mental Method in Medicine, p. 6. Henry Schuman, New York (1952).
