Table Of ContentAaron Polliack
Normal, Transformed
and Leukemic Leukocytes
A Scanning Electron Microscopy Atlas
With 236 Figures
Sp ringer- Verlag
Berlin Heidelberg New York 1977
AARON POLLIACK, M.D.
Associate Professor of Medicine
Department of Hematology
Hadassah University Hospital and Hebrew University
Hadassah Medical School
Jerusalem, Israel
ISBN-13: 978-3-642-66727-5 e-ISBN-13: 978-3-642-66725-1
DOl: 10.1007/978-3-642-66725-1
Library of Congress Cataloging in Publication Data. Polliack, Aaron. 1939- . Normal,
transformed and leukemic leucocytes. Bibliography: p. . Includes index. 1. Leukemia
Atlases. 2. Leukocytes-Atlases. 3. Cell transformation-Atlases. 4. Cancer cells-Atlases.
5. Scanning electron microscope-Atlases. I. Title. RC643.P63 616.1'55'00222 77-14223
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© by Springer-Verlag Berlin Heidelberg 1977.
Softcover reprint of the hardcover I st edition 1977
The use of registered names, trademarks, etc. in this publication does not imply, even in the
absence of a specific statement, that such names are exempt from the relevant protective laws
and regulations and therefore free for general use.
This book is gratefully dedicated to my wife LILY
for her assistance and constant encouragement
Preface
The scanning electron microscope (SEM) has been used with
increasing frequency in recent years to study the surface mor
phology of normal, transformed and malignant leukocytes.
Since the original reports on critical point-dried lymphocytes
published in 1973, results of other studies using improved
methods have been reported giving rise to some controversy
in this field and this is discussed in the text of the atlas.
Advances in preparatory techniques recorded during the past
3 years have also contributed much to a better understanding
of cell surface phenomena as seen under the SEM. The text
of the atlas traces the developments in this field chronologically,
summarizes the available literature and presents the current
situation in the light of the most recent studies in this field.
The photographs were selected to illustrate the spectrum of
surface morphology of the different cell types obtained from
normal individuals and patients with disease states. Hopefully,
the atlas will serve as a guide for future studies and as an
illustration of what SEM has to offer in providing details
of surface architecture.
In an advancing field such as this in which new methods
of cell preparation coupled with vast movements in the resolu
tion of the microscope will no doubt produce better results,
atlases run the risk of becoming outdated quite quickly. Never
theless an overall review needs to be presented at times, to
provide an overview of the current status of knowledge and
of advances in the field, particularly for those who are not
intimately involved in its development. It is hoped that this
atlas fulfills some of these needs and illustrates the exciting
progress which has taken place in recent years.
Jerusalem, Israel AARON POLLIACK
September 1977
Acknowledgments
Some of the studies reviewed and illustrated in this atlas were
performed during 1972-1974 while the author was working
at the Memorial Sloan-Kettering Cancer Center, New York
under the guidance of Etienne de Harven, M.D. and in close
collaboration with Nina Lampen, B.A. Some of the photo
graphs appearing in this album, taken in that laboratory by
Nina Lampen and myself, have been published in earlier re
ports by us as a group and have been reproduced with the
kind permission of the various journals concerned. The author
is grateul to Etienne de Harven and Nina Lampen for their
invaluable assistance during that period. I am also grateful
to Drs. B.D. Clarkson, R.A. Good and H.G. Kunkel for
their constant encouragement, advice and stimulation at the
time of these studies. The skillful photographic assistance of
Juan Marchese during that period is also acknowledged.
Since 1974 these studies have been continued by myself
independently at the Department of Hematology, Hadassah
University Hospital and Hebrew University-Hadassah Medical
School, Jerusalem, Israel. The author is most grateful for the
assistance given him during this period by Professor G.Izak,
M.D. whose constant encouragement has been a source of
stimulation for me. The devoted technical assistance of Miriam
Froimovici is also acknowledged. The author is grateful to
Mr. Moshe Rosenberg of the Medical Photography Depart
ment of the Hebrew University-Hadassah Medical School for
preparation of many of the photographs appearing in this
atlas. The author is also grateful to Shirley Ben Yosef for
devoted secretarial assistance.
The following journals provided permission to reproduce
published figures: The American Journal of Medicine, Biomedi
cine, Blut, British Journal of Haematology, Clinical Immunology
and Immunopathology, Israel Journal of Medical Sciences, Jour
nal of Experimental Medicine, Laboratory Investigation, Scan
dinavian Journal of Haematology. Dr. Om Johari, IIT Research
Institute, Chicago, provided permission for the reproduction
of figures previously published in the proceedings of the SEM
symposia of 1975 and 1976.
Contents
A. Introduction and Methods (Figs. 1 - 8)
B. Review and General Details of the Techniques
Employed to Prepare Leukocytes . . . . . . 5
C. Normal Leukocytes . . . . . . . . . . . . 8
l. Monocytes and Granulocytes (Figs. 9 - 61) 8
2. Lymphocytes and Thymic Cells: Review of the
Factors Affecting the Surface Morphology
of Lymphocytes (Figs. 62 - 91) . . . . . . . 32
D. Labeling Studies on Lymphocytes (Figs. 92 -104) 53
E. Other Factors Influencing the Surface Morphology
of Lymphocytes and Mammalian Cells . . . . .. 63
F . Mitogen-Transformed Lymphocytes (Figs. 105 -124). 64
G. Cultured Cell Lines (Figs. 125 - 158) . . . . . .. 76
H. Surface Features of Leukemic Cells (Figs. 159 - 236) 97
References . 133
Subject Index 139
A. Introduction and Methods
In recent years the scanning electron microscope (SEM) has been
used to study the surface morphology of a variety of mammalian
cells. This microscope has a great depth of focus, provides a three
dimensional view of cell surface topography and enables examination
of a large number of cells. Thus, SEM provides an overall impression
of the surface features of a given population of cells and at the same
time details of individual cell surfaces can be obtained. An added
advantage of this mode of microscopy is that cells can be prepared
relatively quickly for examination in the SEM and results can be ob
tained within a day if necessary. Much, however, depends on the
method of cell preparation and in particular, the modes of fixation
and drying employed in the processing of cells for SEM. Earlier studies
on leukocytes by a number of workers [9, 15, 16, 34, 54] employing
air-drying techniques, did not provide optimal details of surface mor
phology. Air-drying causes cell distortion and shrinkage with collapse
of surface microprojections [10, 82], making it difficult to record details
of the surface morphology of the different cell types and to attempt
to characterize them on the basis of their structure. Nevertheless,
the results of some of the above studies [9, 15, 16, 34, 54] suggested that
lymphocytes had a different surface morphology than monocytes and
that mitogen-transformed lymphocytes differed from normallympho
cytes [28, 34]. However, details of fine surface structure were lacking.
In 1972, Porter [82] and Boyde [10] and co-workers showed that
when cells were processed using freeze-drying or critical point-drying,
results were vastly improved. There was better preservation of the
surface structure which enabled one to characterize and recognize
different types of surface microprojections such as filipodia, microvilli,
blebs and ruffled membranes [82]. The critical point-drying technique
described by Anderson in 1951 [5] and later by Cohen et al. in 1968
[17] allows cells to dry without the disturbing effect of surface tension
and the results are almost optimal. However, a degree of cell shrinkage
still occurs, and cytoplasmic cracks and tears are still seen with this
technique, particularly in elongated fragile cells attached to and spread
ing onto various substrates. When different leukocytes were prepared
by air- and critical point-drying methods, respectively, and then com
pared under the SEM, the superiority of the latter technique was
clearly evident [64] (Figs. 1-8) and it was possible, in many cases,
to characterize different leukocyte types more accurately on the basis
of their surface structure.
2 Critical Point-Drying vs. Air-Drying
4
Figs. 1--4. Critical point-dried cultured lymphoblastoid cells (right) compared with the
same cells which were air-dried (left) showing preservation of surface microprojections
on critical point-dried cells (x 4500-7600)
Critical Point-Drying vs. Air-Drying 3
iiiiii~::=:====-
5
Figs. 5 and 6. High-power photograph of critical point-dried monocyte with convoluted
surface and ruffles (above) compared with the same cell type air-dried (below). Note
the shrinkage of the air-dried cell with loss of fine detail (x 16,300)
