Flow Cytometry: New Developments

469 Pages·1993·13.904 MB·English

by  Bruce D. Jensen (auth.), Alain Jacquemin-Sablon (eds.)

Flow Cytometry
New Developments
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Series H: Cell Biology, Vol. 67
Flow Cytometry
New Developments
Edited by
Alain Jacquemin-Sablon
Centre National de la Recherche Scientifique
Laboratoire de Cytometrie
B.P.3
94801 Villejuif, France
Springer-Verlag
Berlin Heidelberg New York London Paris Tokyo
Hong Kong Barcelona Budapest
Published in cooperation with NATO Scientific Affairs Division
Proceedings of the NATO Advanced Study Institute "4th International Course on Flow
Cytometry" held at Villejuif, France, from April 6-10, 1992
ISBN-13:978-3-642-84618-2 e-ISBN-13:978-3-642-84616-8
001: 10.1007/978-3-642-84616-8
Library of Congress Cataloging-in-Publication Data
Flow cytometry : new developments / edited by Alain Jacquemin-Salon.
(NATO ASI series. Series H, Cell biology; vol. 67)
"Proceedings of the NATO Advanced Study Institute "4th International Course on Flow Cytometry" held at
Villejuif. France. from April 6-10. 1992 - T.p. verso. Published in cooperation with NATO Scientific Affairs
Division." Includes bibliographical references and index.
ISBN-13:978-3-642-84618-2
1. Flow cytometry-Congresses. I. Jacquemin-Salon. Alain. II. North Atlantic Treaty Organization. Scientific
Affairs Division. III. NATO Advanced Study Institute" 4th International Course on Flow Cytometry" (1992 : Villejuif.
France) IV: Series. [DNLM: 1. Flow Cytometry-congresses. QH 585.5.F56 F644 1992]
QH585.5.F56F585 1993 574.87'028-dc20
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© Springer-Verlag Berlin Heidelberg 1993
Softcover reprint of the hardcover 1s t edition 1993
Typesetting: Camera ready by authors
31/3145 -5 4 3 210 - Printed on acid-free paper
Acknowledgements
Generous support form the following Institutions and Companies is gratefully
acknowledged :
- the Centre National de la Recherche Scientifique (CNRS) , the Association
pour la Recherche sur Ie Cancer (.ARc, President : Jacques Crozemarie)
- Becton-Dickinson
- Bio-Rad
- Boehringer Mannheim France
- Chemunex (Partee)
- Coherent
- Coultronics
- Cytolab
- Leica
- Ortho Diagnostic Systems
- Sanofi Recherche
- Spectra-Physics
- Zeiss
We also wish to thank :
- Martine Bardissa
- Claude Boucheix
- Sophie Lafosse
- Dominique Laird
- Linda Pritchard
- Arlette Vervisch
for their expert assistance during the whole Course.
FOREWORD
The International Flow Cytometry course, in the NATO Advanced Study Institute Series,
was designed to address three major areas of interest in cell and molecular biology, (a) cell
activation and biological response, (b) membrane-ligand interactions and cell identity and (c)
nuclear components form and function. Data management, expert systems and cell sorting
techniques that are subjects of concern to all aspects of flow cytometry, were also presented.
Cell activation is an early event in physiological processes that elicits a response of a cell to
perform some specialized function. It is known that several cytolosic events are related to cell
activation. The course focused on optical techniques that can be used in flow cytometry to
study the cascade of processes that are involved. One example was demonstrated by the use
of a peptide honnone for studying the activation of smooth muscle cells. The aim was to
detennine whether calcium and mitochondrial responses could be observed that are consistent
wit the interaction between the signal an energy transduction pathways. Methods for detecting
activation-induced changes in intracellular pH and membrane potential were addressed during
the laboratory exercise. Although the process of cell activation is still not well understood,
many of events that occur in tandem can be studied by flow cytometry and new fluorescent
probes have recently been developed for that purpose.
The capability for following the migration of specific cell types through different parts of the
body has recently become available by a process that involves labeling of cell membrane with
fluorescent, non-toxic probes that are not easily degraded. If cells undergo division the
fluorescent label is partitioned between the two daughter cells. In this way flow cytometric
analyses can be used to "track" and follow the proliferative potential and isolation of specific
subpopulations of cells. Attempts to identify the hemapoeitic stem cells in bone marrow by
flow cytometry have been of particular interest. Some succes has been achieved and stem
cells, capable of reviving a functional hemapoeitic system in lethally irradiated mice have been
be obtained by cell sorting. Several new FCM techniques are being developed for obtaining
viable, cell populations for functional assays.
The numerous functional activities that occur in the cell nucleus is response to intra and inter
cellular signals has invoked a large series of flow cytometric techniques. Changes in
chromatin structure in response to cell cycle or physiological-related changes can be analyzed
with metachromatic fluorochromes such as acridine orange following acid or heat
denaturation of cells. Rates of DNA synthesis are also examined by analysis of BrdU
incorporation into DNA and subsequent flow cytometric analysis of cellular BrdU content
using fluorescein-labeled antibodies to BrdU. Chromosome analysis and sorting of
chromosomes from metaphase arrested cells are now possible. Techniques are also available
to perform fluorescence in-situ hybridization (FISH) on sorted chromosomes. Applications
of PCR and gene mapping on sort chromosomes are now possible. The advanced flow
cytometry course provided the most up to data techniques available at present.
Co-Directors :
Dr. Alain lacquemin-Sablon
CNRS, Laboratoire de Cytometrie, 94800 Villejuif, France
Dr. Harry A Crissman
Los Alamos National Laboratory, Los Alamos, NM 87545, USA
Table of Contents
PART I CELL ACTNATION AND BIOLOGICAL RESPONSE MODIFIERS
page
Cytosolic Events Related to Cell Activation : Potential Interaction Between 3
Signal and Energy Transduction Pathways in Smooth Muscle Cells
B. D. Jensen
Studies of Changes in Cytoplasmic pH and Membrane Potential Using 17
Flow Cytometry
R. E. Worthington and J.P. Aubry
Measurement of Enzyme Kinetics by Flow Cytometry 29
C.Dive, P. Workman, B. L. Marrone and 1 V. Watson
Endocytosis Analysis by Flow Cytometry : Study of the Endocytosis 49
of Fluoresceinylated Neoglycoproteins via Membrane Lectins of Macrophages
P. Midoux, A C. Roche and M. Monsigny
Analysis of Intracellular Antigens by Flow Cytometry : 65
Methods for Cell Permeabilization to Antibodies
H.Giloh
Expression and Role of Surface Markers of Immunocompetent Cells 103
in Non-lymphoid Systems
S. Plaisance, A Alleche, D. Han, E. Rubinstein, Y. Sahraoui, C. Jasmin and
B. Azzarone
Change in Lipid Composition and in Membrane Fluidity of 121
Human Peripheral Blood Lymphocytes undergoing Blastic Transformation
Z. Mishal, A Anel, J.J. Aguilar and 1 Uriel
PART II : MEMBRANE - LIGAND INTERACTIONS AND CELL IDENTITY
Cellular Proliferation History by Fluorescent Analysis 133
P. Horan
Identification and Isolation of Bone Marrow Stem Cells 141
lW.M. Visser and M.G.C. Hogeweg-Platenburg
VIII
page
T.Lymphocytes Recognition Molecules in Homing: 155
A Flow Cytometry Study of Lectin-glycoconjugates Interactions
C. Kieda, N. Bizouarne, V. Denis and M. Mitterand
Three- and Four-color Immunofluorescence Analysis by Flow Cytometry 165
P. Carayon
Energy Transfer 189
W. G. Eisert
Fluid Mechanical Properties of Flow Cytometers and 205
Assessment Cell-cell Adhesion Forces
F. Amblard
Use of Biotin-labeled Growth Factors for Receptor Studies 219
M. O. De long, H. Rozemuller, IG.l. Bauman and 1.W.M. Visser
Cytometrie Tout Terrain or Bush DNA Cytometry 227
S. C. Brown
PART III NUCLEAR COMPONENTS FORM AND FUNCTION
Bromodeoxuridine Procedures for Analysis of DNA Synthesis 245
H. A. Crissman
Flow Cytogenetics : Fundamentals and New Developments 259
L. S. Cram , 1. Fawcett and L.L. Deaven
In situ Hybridization 269
1. A. Bayer and G.Van Den Engh
The Molecular Basis of Mammalian Cell Growth Control 277
G. I. Evan
Flow Cytometric Analysis of Nuclear Associated Antigens 291
1. V. Watson
IX
page
Chromatin Changes Accompanying Apoptosis Detected by Flow Cytometry 311
z. Darzynkiewicz , W. Gorczyca, M.A. Hotz, P. Lassota, S. Bruno and
F. Traganos
Gene Mapping and PCR Applications with Flow-sorted Chromosomes 327
N. P. Carter
Cytometry, Antitumour Drugs and DNA Topoisomerases 343
P.1. Smith
Detection of Apoptosis Using Fluorescent In Situ Nick Translation 355
RR Jonker, 1.G.1. Bauman and 1.W.M. Visser
PART IV DATA MANAGEMENT EXPERT SYSTEMS AND
CELL SORTING TECHNIOUES
Flow Cytometry Data Analysis. Basic Concepts and Analysis of 365
Mono-dimensional Immunofluorescence Histograms
1. V. Watson
Analysis of Multiparameter Flow Cytometric Data 395
J. H. Jett and R C. Habbersett
Expert Assistance for Flow Immunophenotyping 413
G. C. Salzman, R J. Beckman, C. C. Stewart and S. J. Stewart
DNA Content and Cell Cycle Analysis by Flow Cytometry in 421
Clinical Samples : Applications in Cancer
F. Spyratos
Immunomagnetic Particles for Cell Isolation 433
C. Andreoni
Optical Trapping : Instrumentation and Biological Applications 451
T. N. Buican
SUBJECT INDEX 469
PART I
CELL ACTIVATION AND BIOLOGICAL
RESPONSE MODIFIERS