Flow Cytometry Protocols

486 Pages·2011·17.918 MB·English

by  Alice L. Givan (auth.), Teresa S. Hawley, Robert G. Hawley (eds.)

M M B ™
ethods in olecular iology
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK
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Flow Cytometry Protocols
3rd Edition
Edited by
Teresa S. Hawley
Flow Cytometry Core Facility, George Washington University Medical Center, Washington, DC, USA
Robert G. Hawley
Department of Anatomy and Regenerative Biology, George Washington University
Medical Center, Washington, DC, USA
Editors
Teresa S. Hawley Robert G. Hawley
Flow Cytometry Core Facility Department of Anatomy
George Washington University and Regenerative Biology
Medical Center George Washington University
Washington, DC Medical Center
USA Washington, DC
[email protected] USA
[email protected]
ISSN 1064-3745 e -ISSN 1940-6029
ISBN 978-1-61737-949-9 e-ISBN 978-1-61737-950-5
DOI 10.1007/978-1-61737-950-5
Springer New York Dordrecht Heidelberg London
© Springer Science+Business Media, LLC 2011
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Preface
Flow cytometry is riding the crest of unprecedented advances in innovative technologies.
Improvement in instrumentation, lasers, fluorophores, and data analysis software have
facilitated the development of new applications as well as the optimization of existing
applications. This thoroughly revised up-to-date edition highlights the expanding contri-
bution of flow cytometry to basic biological research and diagnostic medicine.
The introductory chapter presents a historical perspective documenting valuable contri-
butions of pioneers in the field. An eloquent synopsis of the principles of flow cytometry
provides a solid foundation for the understanding of basic applications. Modern flow cytom-
etry has been evolving toward high-dimensional complexity. A novel concept, which under-
lies an accurate and efficient strategy for analyzing complex multiparametric data, is
introduced. Great strides have been made toward quantitative fluorescence measurement,
bead-based multiplexed analysis, semiautomated high-throughput flow cytometry, and fluo-
rescence resonance energy transfer the analysis of protein interactions. Other applications
range from polychromatic phenotypic characterizations to genomic and proteomic analyses.
Technologies utilized encompass conventional flow cytometry and imaging cytometry. The
prevalence of aerosol-generating cell/particle sorting warrants a detailed description of stan-
dard safety measures. The last chapter poignantly asserts that while flow and imaging cytom-
etry evolve on a platform of costly sophisticated technologies, minimalist imaging cytometry
holds promise for field-research applications in resource-challenged environments.
The utility of multiparametric flow cytometry is best demonstrated in function-based
studies. Assessment of cytotoxic effector activity and regulatory T cell functions can be
determined using cell tracking dyes, phenotypic markers, and viability probes. Intracellular
cytokine staining is routinely used to visualize antigen-specific T cells. Combined with func-
tional and phenotypic markers, specific cellular subsets can be further dissected. Phospho
flow deciphers intracellular kinase signaling cascades by using highly specific antibodies
which differentiate between the phosphorylated and nonphosphorylated states of proteins.
Combined with immunophenotyping, discrete biochemical signaling events in individual
cells within heterogeneous populations can be carefully examined. The complex progres-
sion of apoptotic death can be evaluated by monitoring multiple apoptotic characteristics
simultaneously. Employing multiple antibodies to detect epitopes on cell cycle-regulated
proteins provides more information than the measurement of DNA content alone.
Single-cell resolution and high-throughput capability make flow cytometry amenable to
the identification of rare cells within a population. Technical aspects of rare event detection
are discussed in the context of practical examples. Direct investigation of distinct cellular
subsets in normal hematopoietic development versus hematologic diseases is critical to the
understanding of disease initiation and progression. High-resolution polychromatic fraction-
ation of hematopoietic precursors dissects developmental stages and identifies cellular subsets
with defined lineage potentials. Carefully devised protocols in the study of human hemato-
logic disorders enable the diagnosis and monitoring of patients with leukemia and lymphoma,
or primary immunodeficiency diseases. Cell-derived microparticles, which are implicated in
pathogenesis, can be analyzed by conventional as well as imaging flow cytometry.
v
vi Preface
The impact of fluorescent proteins (FPs) on bioscience is underscored by the award of
the 2008 Nobel Prize in Chemistry to three scientists involved in the discovery and sub-
sequent optimization of the green fluorescent protein (GFP). Unlike other biolumines-
cent reporters, fluorescence of GFP and GFP-related proteins does not require exogenous
substrates or cofactors. GFP from jellyfish and GFP-like proteins from other marine organ-
isms, as well as recently engineered nontoxic red-shifted variants, have played vital roles in
the noninvasive detection of genes transferred into living cells. Several schemes for the
simultaneous detection of multiple FPs in living cells were presented in the second edi-
tion. The applications described in this edition further illustrate the broad utility of FPs in
flow cytometry. The study of plant biology has been accelerated by the use of FP expres-
sion constructs. Other downstream applications involve genome-wide expression profiling
and mass spectrometry, two analysis platforms that readily interface with flow cytometry.
Such integration bodes well for the study of systems biology.
We would like to thank John Walker for inviting us to participate in this exciting
endeavor again and for his expert editorial guidance. We are especially grateful to all of the
contributors for their enthusiasm and generosity. Their willingness to impart their knowl-
edge exemplifies the spirit of cooperation that is pervasive in the cytometry community.
As the “Notes” section is a hallmark of this series, we are proud to present chapters that
contain up to 59 notes!
Washington, DC Teresa S. Hawley
Washington, DC Robert G. Hawley
Contents
Preface............................................................. v
Contributors......................................................... ix
1 Flow Cytometry: An Introduction .................................... 1
Alice L. Givan
2 Breaking the Dimensionality Barrier................................... 31
C. Bruce Bagwell
3 Quantitative Fluorescence Measurements
with Multicolor Flow Cytometry ..................................... 53
Lili Wang, Adolfas K. Gaigalas, and Ming Yan
4 Quantum Dots for Quantitative Flow Cytometry ......................... 67
Tione Buranda, Yang Wu, and Larry A. Sklar
5 Bead-Based Multiplexed Analysis of Analytes by Flow Cytometry ............. 85
Henri C. van der Heyde and Irene Gramaglia
6 Flow-Based Combinatorial Antibody Profiling:
An Integrated Approach to Cell Characterization ......................... 97
Shane Bruckner, Ling Wang, Ruiling Yuan, Perry Haaland,
and Amitabh Gaur
7 Tracking Immune Cell Proliferation and Cytotoxic
Potential Using Flow Cytometry...................................... 119
Joseph D. Tario Jr., Katharine A. Muirhead, Dalin Pan,
Mark E. Munson, and Paul K. Wallace
8 Multiparameter Intracellular Cytokine Staining........................... 165
Patricia Lovelace and Holden T. Maecker
9 Phospho Flow Cytometry Methods for the Analysis of Kinase
Signaling in Cell Lines and Primary Human Blood Samples ................. 179
Peter O. Krutzik, Angelica Trejo, Kenneth R. Schulz,
and Garry P. Nolan
10 Multiparametric Analysis of Apoptosis by Flow Cytometry .................. 203
William G. Telford, Akira Komoriya,
Beverly Z. Packard, and C. Bruce Bagwell
11 Multiparameter Cell Cycle Analysis.................................... 229
James W. Jacobberger, R. Michael Sramkoski, and Tammy Stefan
12 Rare Event Detection and Analysis in Flow Cytometry:
Bone Marrow Mesenchymal Stem Cells, Breast Cancer
Stem/Progenitor Cells in Malignant Effusions,
and Pericytes in Disaggregated Adipose Tissue ........................... 251
Ludovic Zimmerlin, Vera S. Donnenberg, and Albert D. Donnenberg
vii
viii Contents
13 Flow Cytometry-Based Identification of Immature
Myeloerythroid Development........................................ 275
Cornelis J.H. Pronk and David Bryder
14 Flow Cytometry Immunophenotyping of Hematolymphoid Neoplasia ...... 295
Katherine R. Calvo, Catharine S. McCoy, and Maryalice Stetler-Stevenson
15 Flow Cytometry Assays in Primary Immunodeficiency Diseases............... 317
Maurice R.G. O’Gorman, Joshua Zollett, and Nicolas Bensen
16 Flow Cytometric Analysis of Microparticles ............................. 337
Henri C. van der Heyde, Irene Gramaglia, Valéry Combes,
Thaddeus C. George, and Georges E. Grau
17 Noncytotoxic DsRed Derivatives for Whole-Cell Labeling................... 355
Rita L. Strack, Robert J. Keenan, and Benjamin S. Glick
18 Flow Cytometric FRET Analysis of Protein Interaction..................... 371
György Vereb, Péter Nagy, and János Szöllo˝si
19 Fluorescent Protein-Assisted Purification for Gene Expression Profiling ........ 393
M. Raza Zaidi, Chi-Ping Day, and Glenn Merlino
20 Multiparametric Analysis, Sorting, and Transcriptional Profiling
of Plant Protoplasts and Nuclei According to Cell Type................. 407
David W. Galbraith, Jaroslav Janda, and Georgina M. Lambert
21 Lentiviral Fluorescent Protein Expression Vectors
for Biotinylation Proteomics......................................... 431
Irene Riz, Teresa S. Hawley, and Robert G. Hawley
22 Standard Practice for Cell Sorting in a BSL-3 Facility ...................... 449
Stephen P. Perfetto, David R. Ambrozak, Richard Nguyen,
Mario Roederer, Richard A. Koup, and Kevin L. Holmes
23 The Cytometric Future: It Ain’t Necessarily Flow! ........................ 471
Howard M. Shapiro
Erratum............................................................ E1
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 483
Contributors
DaviD R. ambRozak • Vaccine Research Center, National Institute of Allergy
and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
C. bRuCe bagwell • Verity Software House, Topsham, ME, USA
NiColas beNseN • Department of Pathology and Laboratory Medicine,
The Children’s Memorial Hospital, Chicago IL, USA
shaNe bRuCkNeR • BD Biosciences, San Diego, CA, USA
DaviD bRyDeR • Department of Immunology, Institution for Experimental
Medical Science, Lund University, Lund, Sweden
TioNe buRaNDa • Department of Pathology and Cancer Center,
University of New Mexico School of Medicine, Albuquerque, NM, USA
kaTheRiNe R. Calvo • Department of Laboratory Medicine, Hematology Section,
National Institutes of Health Clinical Center, Bethesda, MD, USA
valéRy Combes • Department of Pathology, University of Sydney,
Camperdown, NSW, Australia
Chi-PiNg Day • Laboratory of Cancer Biology and Genetics,
Center for Cancer Research, National Cancer Institute,
National Institutes of Health, Bethesda, MD, USA
albeRT D. DoNNeNbeRg • Division of Hematology/Oncology,
Department of Medicine, University of Pittsburgh School of Medicine
and University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA
veRa s. DoNNeNbeRg • Department of Surgery, University of Pittsburgh
School of Medicine and University of Pittsburgh Cancer Institute,
Pittsburgh, PA, USA
aDolfas k. gaigalas • National Institute of Standards and Technology,
Gaithersburg, MD, USA
DaviD w. galbRaiTh • Department of Plant Sciences, University of Arizona,
Tucson, AZ, USA
amiTabh gauR • BD Biosciences, San Diego, CA, USA
ThaDDeus C. geoRge • Amnis Corporation, Seattle, WA, USA
aliCe l. givaN • Englert Cell Analysis Laboratory of the Norris Cotton
Cancer Center and Department of Physiology, Dartmouth Medical School,
Lebanon, NH, USA
beNjamiN s. gliCk • Department of Molecular Genetics and Cell Biology,
Cummings Life Science Center, The University of Chicago, Chicago, IL, USA
iReNe gRamaglia • La Jolla Infectious Disease Institute, San Diego, CA, USA
geoRges e. gRau • Department of Pathology, University of Sydney,
Camperdown, NSW, Australia
PeRRy haalaND • BD Technologies, Research Triangle Park, NC, USA
ix
x Contributors
RobeRT g. hawley • Department of Anatomy and Regenerative Biology,
The George Washington University Medical Center, Washington, DC, USA
TeResa s. hawley • Flow Cytometry Core Facility, The George Washington
University Medical Center, Washington, DC, USA
heNRi C. vaN DeR heyDe • Cell Analysis Core Facility, Flow Cytometry,
La Jolla Infectious Disease Institute, San Diego, CA, USA
keviN l. holmes • Flow Cytometry Section, Research Technologies Branch,
National Institute of Allergy and Infectious Diseases, National Institutes
of Health, Bethesda, MD, USA
james w. jaCobbeRgeR • Cytometry and Imaging Microscopy Core,
Case Comprehensive Cancer Center, Case Western Reserve University,
Cleveland, OH, USA
jaRoslav jaNDa • Department of Plant Sciences, University of Arizona,
Tucson, AZ, USA
RobeRT j. keeNaN • Department of Biochemistry and Molecular Biology,
Gordon Center for Integrated Sciences, The University of Chicago,
Chicago, IL, USA
akiRa komoRiya • OncoImmunin, Inc ., Gaithersburg, MD, USA
RiChaRD a. kouP • Vaccine Research Center, National Institute of
Allergy and Infectious Diseases, National Institutes of Health,
Bethesda, MD, USA
PeTeR o. kRuTzik • Baxter Laboratory in Stem Cell Biology,
Department of Microbiology and Immunology, Stanford University,
Stanford, CA, USA
geoRgiNa m. lambeRT • Department of Plant Sciences, University of Arizona,
Tucson, AZ, USA
PaTRiCia lovelaCe • Institute for Immunity, Transplantation, and Infection,
Stanford University, Stanford, CA, USA
holDeN T. maeCkeR • Human Immune Monitoring Center, Institute
for Immunity, Transplantation, and Infection, Stanford University,
Stanford, CA, USA
CaThaRiNe s. mCCoy • Flow Cytometry Laboratory, Laboratory of Pathology,
National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
gleNN meRliNo • Laboratory of Cancer Biology and Genetics,
Center for Cancer Research, National Cancer Institute,
National Institutes of Health, Bethesda, MD, USA
kaThaRiNe a. muiRheaD • SciGro, Inc ., Madison, WI, USA
maRk e. muNsoN • Verity Software House, Topsham, ME, USA
PéTeR Nagy • Department of Biophysics and Cell Biology,
Research Center for Molecular Medicine, Medical and Health
Science Center, University of Debrecen, Debrecen, Hungary
RiChaRD NguyeN • Vaccine Research Center, National Institute of Allergy
and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
gaRRy P. NolaN • Baxter Laboratory in Stem Cell Biology,
Department of Microbiology and Immunology, Stanford University,
Stanford, CA, USA