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Practical Flow Cytometry

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ISBN-10: 0471411256

ISBN-13: 9780471411253

Edition: 4th 2003 (Revised)

Authors: Howard M. Shapiro

List price: $221.50
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This complete reference provides the background science and discussion of the vast biomedical applications of quantitative analytical cytology using laser-activated detection and cell sorting.
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Book details

List price: $221.50
Edition: 4th
Copyright year: 2003
Publisher: John Wiley & Sons, Incorporated
Publication date: 7/14/2003
Binding: Hardcover
Pages: 736
Size: 8.60" wide x 11.10" long x 1.80" tall
Weight: 4.158
Language: English

List of Tables and Figures
Preface to the Fourth Edition: Why you Should Read this Book - or Not
Foreword to the Third Edition
Preface to the Third Edition
Preface to the Second Edition
Foreword to the First Edition
Preface to the First Edition
What (And What Good) Is Flow Cytometry?
Tasks and Techniques of Cytometry
Some Notable Applications
What is Measured: Parameters and Probes
Beginnings: Microscopy And Cytometry
A Little Light Music
Making Mountains out of Molehills: Microscopy
Why Cytometry? Motivation and Machinery
Flow Cytometry and Sorting: Why and How
Problem Number One: Finding The Cell(s)
Flow Cytometry: Quick on the Trigger
The Main Event
The Pulse Quickens, the Plot Thickens
Flow Cytometry: Problems, Parameters, Probes, and Principles
Counting Cells: Precision I (Mean, S.D., CV)
And Now to See with Eye Serene the Very Pulse of the Machine: Display, Digitization, and Distributions
DNA Content Analysis: Precision II (Variance)
Two-Parameter Displays: Dot Plots and Histograms
Identifying Cells in Heterogeneous Populations: Lift Up Your Heads, Oh Ye Gates!
Sorting Sorting Out
Parameters and Probes II: What is Measured and Why
What's In the Box: Flow Cytometer Anatomy, Physiology, and Pathology
Light Sources for Microscopy and Flow Cytometry
Instrument Configurations: The Orthogonal Geometry
Laser Beam Geometry and Illumination Optics
Flow Chamber and Forward Scatter Collection Optics
Fluorescence and Orthogonal Scatter Optics
Optical Filters for Spectral Separation
Multistation Flow Cytometers
Photomultipliers and Detector Electronics
Putting the Flow in Flow Cytometry
Signal Processing Electronics
Is It Bigger than a Breadbox?
Flow Cytometer Pathology and Diagnostics
Alternatives to Flow Cytometry; Cytometer Ecology
The Rest Of The Book
Lis(z)t Mode
Learning Flow Cytometry
Learning from History: Take One
Who Should Read this Book?
Information Sources and Resources
Books on Flow Cytometry in General
Books on Flow Cytometric Methodology and Protocols
Clinical Flow Cytometry Books
Other Flow Cytometry Books
Flow's Golden Oldies
The Reader's Guide To Periodical Literature
Resources And Courses
Flow Cytometer Manufacturers
The International Society for Analytical Cytology
The Clinical Cytometry Society
The National Flow Cytometry Resource
"The Annual Courses" and Others
Other Societies and Programs
The Purdue Mailing List, Web Site, and CD-ROMs
Exploring The Foundations
Optics and Microscopy
Computers: Hardware and Software
Digital Signal Processing
Data Presentation and Display
Spectroscopy, Fluorescence and Dye Chemistry
Cell and Molecular Biology and Immunology
Alternatives To Flow Cytometry
Ancient History
Flow Cytometry: Conception and Birth
Staining Before and After Paul Ehrlich
Origins of Modern Microscopy
Making Cytology Quantitative: Caspersson et al
Origins of Cancer Cytology: The Pap Smear
The Fluorescent Antibody Method
Blood Cell Counting: Theory and Practice
Video and Electron Microscopy
Optical Cell Counters and the Coulter Orifice
Classical History
Analytical Cytology in the 1950's
The Cytoanalyzer
Acridine Orange as an RNA Stain: Round One
How I Got Into this Mess
The Rise of Computers
Computers in Diagnosis: A Central Problem
Diagnosis and Classification: Statistical Methods
Cytology Automation in the 1960's
First Steps toward Automated Differentials
Pattern Recognition Tasks in Cell Identification
Differential Leukocyte Counting: An Early Flow Systems Approach
Kamentsky's Rapid Cell Spectrophotometer
Fulwyler's Cell Sorter
Modern History
Cell Cycle Analysis: Scanning versus Flow Systems
Cancer Cytology: Scanning versus Flow Cytometry
Early Commercial Flow Cytometers
Not Quite Commercial: The Block Projects
The Evolution of Flow Cytometers in the 1970's
Dog Days: The Genesis of Cytomutts
The 1980's: Little Things Mean a Lot
Measurements in the Main Stream
Clinical Uses of Fluorescence Flow Cytometry
The End of History?
How Flow Cytometers Work
Light and Matter
Photometry versus Radiometry: What's in a Name?
Physical Measurement Units
Light in Different Lights
It's All Done With Photons
A Few Warm Bodies
Polarization and Phase; Interference
Light Meets Matter: Rayleigh and Mie Scattering
A Time for Reflection-and Refraction: Snell's Law
Polarization by Reflection; Brewster's Angle
Dispersion: Glass Walls May Well a Prism Make
Interference in Thin Films
Interference and Diffraction; Gratings
Optical Activity and Birefringence
Matter Eats Light: Absorption
Absorption: Counting the Calories
A Selective Diet
The Chance of a Lifetime
Spinning a Tale of Degeneracy
Facing Extinction: Cross Section and Optical Density
Unexciting Times: Emigrating from the Excited States
Fluorescence: Working the Stokes Shift
Fluorescence Polarization
Stimulated Emission
Resonance Energy Transfer
Quenching, Bleaching, and Photon Saturation
Quantum Flotsam and Jetsam
Optical Systems
Light Propagation and Vergence
Image Formation by Optical Systems: Magnification
Lens Types and Lens Aberrations
Numerical Aperture and Lens Performance
Gradient Index, Fresnel, and Cylindrical Lenses
The Helmholtz Invariant and Throughput
Photons in Lenses: See How They Run
Aperture and Field Stops: The f Number
Depth of Field and Focus and Resolution of Lenses
Light Sources
The Best and the Brightest
Harc, Harc, the Arc!
Quartz Halogen Lamps
Light Emitting Diodes (LEDs)
Illumination Optics for Lamps and LEDs
Arc Source Epiillumination for Flow Cytometry
Lasers as Light Sources for Flow Cytometers
Laser Illumination: Going to Spot
Shedding Light on Cells: Lasers, Lamps, and LEDs
Lasers: The Basic Physics
Lasers Used and Usable in Cytometry
Laser and Light Source Noise and Noise Compensation
Danger!!! Laser!!! Hazards and Haze
Light Collection
Microscope Objectives
Looking at the Observation Point
Stops versus Blockers
Signal versus Noise: To See or Not to See
Spectral Selection: Monochromators versus Filters
Neutral Density Filters
Beamsplitters; Ghosts and Ghostbusters
Optics for Polarization Measurements
Tunable Filters
Fiber Optics and Optical Waveguides
Through a Glass Darkly: Light Lost (and Found) in Optical Components
Collection Optics for Forward Scatter Signals
Silicon Photodiodes
Photomultiplier Tubes (PMTs)
Sensitivity Training: Photodiode versus PMT
Single Photon Counting
Avalanche Photodiodes (APDs)
PMTs: Picking a Winner
Photomultipliers: Inexact Science
Charge Transfer Devices: CCDs, CIDs, Etc.
Flow Systems
Flow System Basics
When You've a Jet
Core and Sheath: Practical Details
Grace Under Pressure: Driving the Sheath and Core
Perfect Timing: Fluidics for Kinetic Experiments
Oriented and Disoriented Cells
Matchmaker, Matchmaker, Make Me a(n) Index Match!
Flow Unsheathed
Flow Systems: Garbage In, Garbage Out
Electronic Measurements
Electricity and Electronics 101
The Coulter Principle: Electronic Cell Sizing
Analog Signal Processing
Beam Geometry and Pulse Characteristics
Electronics 102: Real Live Circuits
Detector Preamplifiers and Baseline Restoration
Analog Pulse Processing: Front Ends and Triggering
Dead Times, Doublets, and Problem Pulses
Trigger Happy?
Analog Linear, Log and Ratio Circuits
Digital Signal Processing
Analog-to-Digital Conversion
Digital Pulse Processing and DSP Chips
Digitization: Tying it All Together
Performance: Precision, Sensitivity, and Accuracy
Precision; Coefficient of Variation (CV)
Minimum Detectable Signal
MESF Units
Linearity and Nonlinearity
What's All the Noise About?
More Photons Give Better Precision
Background Effects
Electrons Have Statistics, Too
Source Noise Fluctuations and Performance
I Blurred It Through the Baseline
Restoration Comedy: The Case of the Disappearing Leukocytes
Top 40 Noise Sources
Sensitivity 007: Q and B (Dye Another Day?)
Data Analysis
Goals and Methods in Data Analysis
Cell Counting
Characterization of Pure Cell Populations
Identification of Cells in Mixed Populations
Characterization of Cell Subpopulations
Data Analysis Hardware and Software Evolve
Computer Systems for Flow Cytometry
The Beginning
The End of the Beginning
Data Rates and Data Acquisition Systems
Primary Data: Frequency Distributions
You Say You Want a Distribution
Distributions Have Their Moments
Some Features of the Normal Distribution
Measures of Central Tendency: Arithmetic and Geometric Means, Median, and Mode
Measures of Dispersion: Variance, Standard Deviation, CV, and Interquartile Range
Robustness in Statistics; the Robust CV
Calculating and Displaying Histograms
Bivariate and Multivariate Distributions and Displays
Compensating Without Decompensating
Dealing With the Data
Comparing and Analyzing Univariate Histograms
Deconvoluting Single-Parameter Histograms
Analysis of Two-Parameter Data
Analysis of Two-Parameter Distributions
Multiparameter Data Analysis
Multiparameter versus Multivariate Analysis
Multiparameter Analysis of Leukocyte Types: 1974
Multiparameter Analysis of Leukocyte Types: 2002
Procedures for Automated Classification
Analysis of Collected Data: How Much Is Enough/Too Much?
Data Analysis Odds and Ends
Data Storage
Linear and Log Scales and Ratios: Proceed with Care!
Flow Sorting
Sort Control (Decision) Logic
Preselected Count Circuits and Single Cell Sorting
Droplet Sorting, High-Speed and Low
Droplet Generation
Drop Charging and Deflection
Determining Droplet Delay Settings
Transducers and Transducer Drive Signals
Improving Droplet Sorting
Sorting Large Objects with Droplet Sorters
Fluidic Switching Cell Sorters
Sorting Large Objects Using Fluidic Switching
Sorting Very Small Objects: Microfluidic Switching
Cell Manipulation By Optical Trapping
Cell Damage Cell Selection ("Cell Zapping")
Photodamage Cell Selection
Sorting (Zapping) Without Flow (Gasp!)
Electrodamage Cell Selection in Flow
Measures of Cell Sorter Performance: Purity, Recovery (Yield), and Efficiency
Coincidence Effects on Performance
Other Considerations
Doing the Math
Speed Limits: The Reynolds Rap
Instrument Utilization
Monitoring versus Sorting for Cell Preparation
Collection Techniques: Life and Death Decisions
Dilutions of Grandeur
Can Getting Sorted Be Hazardous to Cells' Health?
Biohazard Control and Biosafety in Flow Cytometers and Sorters
Parameters and Probes
Physical Parameters and Their Uses
Electrical Parameters
Acoustic Measurements of Cells in Flow
Optical Parameters: Light Scattering
Optical Parameters: Absorption
Optical Parameters: Extinction
Other Transmitted Light Measurements
Optical Parameters: Fluorescence
Optical Parameters: Fluorescence
Intrinsic Cellular Parameters
Cell Size
Cell Shape and Doublet Discrimination
Measurement of Intrinsic Parameters Using Absorption and Extinction Signals
Fluorescence Measurement of Intrinsic Parameters
Probes, Labels, and [Not] Protocols for Extrinsic Parameter Measurements
Probes, Labels, and Dyes
Dyes and Quality Control: Gorillas in the NIST
The Dyes are Cast: An Overview
Mechanisms of Staining by Fluorescent Dyes
"Vital" Staining
Fixation--Why and How
Red Blood Cell Lysis: The Distilled Essence
Nucleic Acid Dyes and Their Uses
DNA Content Measurement
DNA Base Composition
Chromatin Structure; Identifying Cells in Mitosis
RNA Content
Fluorescent Labels and Protein Dyes
Estimating Total and Basic Protein Content of Cells
Covalent Labels for Antibodies and Other Molecules
Future Tandems: Heterocycles Built for Two?
Cyanine Dye Labels: From Cy-Fi to Hi5 for Cy5
Blue Notes: AMCA and Cascade Blue
Alexa Dyes: Some Thoughts on Dyemographics
Other Organic Fluorescent Labels: A Dye Named Joe, etc.
Quantum Dots
Getting Labels Onto Molecules of Interest
Improving Signals from Labels: Amplification and Other Techniques
Limits to Sensitivity: Autofluorescence
Raman Scattering Effects on Sensitivity
Increasing Sensitivity: Amplification Techniques
Improving Sensitivity: Time-Resolved Fluorescence
Measuring Cell Surface and Intracellular Antigens
History and Background
Antibody Reagents and Staining Procedures
Fluorescence Measurements: Lurching Toward Quantitation
Nucleic Acid Sequence Detection
Peptide Nucleic Acid (PNA) Probes
Probes for Various Cell Constituents
Surface Sugars (Lectin Binding Sites
Analysis of Total Carbohydrate Content
Specific Detection of Cellulose
A Probe for Cell Surface Aldehydes
Probes for Lipids and Cholesterol
Probes for Cytoskeletal Organization/Actins
Time as a Parameter: Kinetic Measurements
Sample Handling for Kinetic Measurements
Time as a Quality Control Parameter
Slooowww Flooowww
Labeled Ligand Binding
Labeling Strategies
Formal Analysis of Ligand binding
Labeled Ligands versus Anti-Receptor Antibodies
Ligand Binding Detected by Functional Changes
Fluorescent Ligand Binding: Some Examples
Functional Parameters I
Cell Surface Charge
Cell Membrane Characteristics
Enzyme Activity
Sulfhydryl (Thiol) Groups; Glutathione
Functional Probes II: Indicators of Cell Activation
Changes in the Cellular Ionic Environment Following Activation by Ligand Interaction with Cell Surface Receptors
"Structuredness of Cytoplasmic Matrix" (SCM) and the Cercek Test for Cancer
Optical Probes of Cell Membrane Potential
Optical Probes of Intracellular Calcium
Flow Cytometric Probes of Intracellular pH
The Hat Trick: Multiparameter Approaches to Ion Flux Measurements in Cell Activation
NOsing Around for Nitric Oxide
Other Ions in the Fire
Reporter Genes
Somebody Cloned My Gal: Enzymes as Reporter Genes
Green Fluorescent Protein (GFP) et al
Minority Report(er)?
Buying Flow Cytometers
BD Biosciences
The BD FACS Vantage SE Cell Sorter
The BD FACSCalibur Analyzer
The B-D LSR II Analyzer
The B-D FACSAria Cell Sorter
The B-D FACSCount
Beckman Coulter, Inc.
Background--and Signal-to-Background
The Beckman Coulter EPICS ALTRA Cell Sorter
The Beckman Coulter Cytomics FC 500 Analyzer
The EPICS XL and XL-MCL Analyzers
The MoFlo Cell Sorter
The CyAn Flow Cytometer
The InFlux Cell Sorter
Optoflow AS
The MICROCYTE Flow Cytometer
Partec GmbH
The CyFlow and CyFlow ML Flow Cytometers
The PAS, PAS II, and PAS III Flow Cytometers
PA Ploidy Analyzer and CCA Cell Counter Analyzer
Some Other Flow Cytometer Companies
Advanced Analytical Technologies, Inc. (AATI)
Agilent Technologies, Inc.
Apogee Flow Systems Ltd.
Bentley Instruments
Chemunex SA
CytoBuoy b.v.
Delta Instruments bv
Fluid Imaging Technologies, Inc.
FOSS Electric A/S
Guava Technologies, Inc.
Howard M. Shapiro, M.D., P.C.
iCyt--Visionary Bioscience
International Remote Imaging Systems
Luminex Corporation
NPE Systems, Inc.
Union Biometrica, Inc.
Hematology Instruments, Etc.
Little Orphan Analyzers (And Big Orphan Sorters)
Bio/Physics and Ortho: Cytofluorograf to Cytoron
HEKA Elektronik GMBH: The FLUVO II Analyzer
The Kratel Partograph
The ODAM ATC 3000
Also Among the Missing
Flow Cytometer Rehabilitation; Used Instruments
Following Suit
Third Party Software
The Selling of Flow Cytometers: Hype and Reality
Applying for a Grant for a Cytometer
Building Flow Cytometers
Why Buy a Flow Cytometer?
Why Build a Flow Cytometer?
Learning to Build Your Own
Using Flow Cytometers: Applications, Extensions, and Alternatives
The Daily Grind
Keeping the Instrument Running: Diet and Exercise
Particulars: Drawing a Bead on Flow Cytometer Alignment, Calibration, and Standardization
Rose Colored Glasses: Optical Filter Selection
Experimental Controls
Shake Well Before Using: When Controls Won't Help
Significant Events in the Lives of Cells
Taking the Census: Cell Counting
The Doubled Helix: Reproduction
Memento Mori: Detecting Cell Death
Die Another Day: Cytometry of Telomeres
Identification of Cells in Mixed Populations
Mixed Genotypes versus Mixed Phenotypes
No Parameter Identifies Cancer Cells
Many Parameters Identify Blood Cells
Flow Cytometric Parameters Useful for Blood Cells
Specific Gene Products Identify Cell Types
Maturation Processes and "Missing Links": The "Ginger Root" Model
Practical Multiparameter Gating: Color Wars
Finding Rare Cells
Tricks and Twists: Odd Jobs for Flow Cytometry
Single Molecule Detection
DNA Sizing, if not Sequencing, in Flow
Solid Phase (Bead) Assays Using Flow Cytometry
Cells in Gel Microdroplets and on Microspheres
Single Cell Analysis: When Flow Won't Do
Applications of Flow Cytometry
Cell Differentiation, Ab Ovo and De Novo
Somatic Cell Genetics and Cell Hybridization
Chromosome Analysis and Sorting and Flow Karyotyping
Probing Details of Cellular Structures and Inter- and Intramolecular Interactions
Clinical Flow Cytometry: Turf and Surf
Cancer Biology and Clinical Oncology
Analysis of Sperm
Isolating Fetal Cells from the Maternal Circulation for Prenatal Diagnosis
The March of Time: Circadian Rhythms, Aging, and Atherosclerosis
Clinical Application: Urine Analysis
The Animal Kingdom
Big Stuff, Vegetable, Animal or Mineral
Flow Cytometry of Plant Cells and Chromosomes
Microbiology, Parasitology and Marine Biology
Pharmacology and Toxicology
Food Science
Biotechniques and Biotechnology
Alternatives: Microfluidic Cytometers, Flow and Static
Cytometry Afield
Sources of Supply
Resources, Societies, Journals
Optical Supply Houses
Probes and Reagents
Calibration Particles/Cytometry Controls
Flow Cytometers
Hematology Instruments
Data Analysis Software/Systems
Hardware and Software
Commercial Software Sources
Noncommercial Software Sources
Cytometer Rehabilitation/Add-ons
Flow Cytometer Parts
Flow System Plumbing
DC-DC Converter Modules for HV Power Supplies
Power Supplies (Low Voltage)
Other Electronics
Laser Trade Publications
Laser Manufacturers
Optical Filters
Color Glass Filters
Interference Filters
Neutral Density Filters
Polarizing Filters and Optics
Tunable Filters
Aids to Troubleshooting Flow Cytometers When All Else Fails
Proficiency Testing
Sex Selection
Alternative Technology
Dotting i's and Crossing t's
Late Breaking News
New Book
New Protein Stain
Caveat on Fluorescent Caspase Inhibitors
Polyamide Probes
Tearing Down the (Picket) Fences
New Instrument: The BD FACSArray
Science Special Section: Biological Imaging
Cytomics in Predictive Medicine: a Clinical Cytometry Special Issue and Other Recent Citings and Sightings
Analytical Biology, Such as it Isn't: Is This Any Way to Run a Science?
Unfinished Business
AIDS and Infectious Disease in the Third World
A Center for Microbial Cytometry
A Nobel Prize for Herzenberg and Kamentsky?
Flow and the Human Condition
There's No Business Like Flow Business
One More Thing