TABLE OF CONTENTS

INTRODUCTION: A Palette of Living Colors; Rochelle Diamond

HISTORY: Susan DeMaggio

GETTING STARTED

• Chapter 1: How the FACScalibur and FACSvantage Work and Why It Matters; Tom Frey, Ph.D.
• Chapter 2: How Flow Cytometers Work and Don’t Work; Howard M. Shapiro, M.D., P.C.
• Chapter 3: Flow Cytometry Standard (FCS) Data File Format; Larry Seamer
• Chapter 4: Data Display; Susan DeMaggio and Rochelle Diamond
• Chapter 5: Guidelines to Improve Flow Cytometry Data Display and Interpretation;Wendy Schober and Dorothy Lewis
• Chapter 6: Data Management and Storage; Dave Coder
• Chapter 7: Coloring up: A guide to Spectral Compensation; Rochelle Diamond
• Chapter 8: Consistency in Quality Control; Rochelle Diamond

SAMPLE PREPARATION AND CELL SURFACE STAINING

• Chapter 1: Cell Preparation and Enrichment for FCM Analysis and Cell Sorting; Rochelle Diamond, Hua Wang, Fei Chen, and Mindy Wilke-Douglas
• Chapter 2: The Basics of Staining for Cell Surface Proteins; Alice Givan
• Chapter 3: Phenotypic Analysis Using Five-Colour Immunofluorescence and Flow Cytometry; Andrew Beavis
• Chapter 4: Detection of Cell Surface Antigens Using Antibody-Conjugated Fluorospheres; Andrew Beavis
• Chapter 5: Gel Microdrop Encapsulation for the Frugal Investigator; David McFarland and Gary Durack

REPORTER GENES AND CELL TRACKERS

• Chapter 1: Flow Cytometric Analysis of GFP Expression in Mammalian Cells; Steven R. Kain, Ph.D.
• Chapter 2: FACS-Gal: Flow Cytometric Assay of B-galactosidase; Steve Fiering
• Chapter 3: Bicistronic Vectors and FCM; Nicholas Poulos, Eric Stanbridge, and Susan DeMaggio
• Chapter 4: NGFR as a Reporter Gene; G. Raghu, M. Munn, S.W. Park, C-H Pan, and A. Dayn
• Chapter 5: Assessment of Cell Migration in vivo Using the Fluorescent Tracking Dye PKH26 and Flow Cytometry; Andrew Beavis
• Chapter 6: In vivo Biotinylation for Cell Tracking and Survival/Lifespan Measurments; Kenneth A. Ault, M.D. and Cathy Knowles
• Chapter 7: MACSelect Chapter – Title still coming; Mindy Wilke-Douglas and Ute Behrens-Jung
• Chapter 8: Cell Tracking Using PKH; Rebecca Poon and Bruce Bagwell

FLOW CYTOMETRY OF NUCLEIC ACIDS

• Chapter 1: RNA Content Determination Using Pyronin Y; Howard Shapiro, M.D., P.C.
• Chapter 2: DNA Content Determination; Derek Davies
• Propidium Iodide Staining for DNA Content
• Vital Staining for DNA with Hoechst 3342
• Bromodeoxyuridine Staining for Cell Cycle Kinetics
• Chapter 3: Dead Cell Discrimination; Derek Davies
• Propidium Iodide Staining for Dead Cell Discrimination
• Dead Cell Discrimination Using ToPro-3
• Chapter 4: Detecting Apoptosis; Derek Davies & Ingrid Schmid
• Hoechst/Propidium Iodide Detection for Apoptotic Cells
• 7-AAD Staining for Apoptosis
• Annexin-FITC Staining for Apoptotic Cells
• Chapter 5: Chromosome Preparation; Derek Davies and Clare Hughes
• Chapter 6: D-Flowering – The Flow Cytometry of Plant DNA; John Mottley, K. Yokoya, A.V. Roberts
• Chapter 7: Flow Cytometry Analysis of Marine Picoplankton; Dominique Marie, N. Simon, L. Guillon, F. Partensky, D. Vaulot
• Chapter 8: Measuring Yeast DNA; Wendy Schober and Jeff Bachant

INTRACELLULAR PHYSIOLOGICAL AND ANTIBODY PROBES

• Chapter 1: Calcium Flux in INDO-1 Stained Thymocytes; Geoffrey W. Osborne and Paul Waring, Ph.D.
• Chapter 2: Measuring Intracellular pH Using ADB; Mike Fox
• Chapter 3: Measuring Intracellular pH Using SNARF-1; Mike Fox
• Chapter 4: SNARF-1 and Apoptosis; Judy Norberg, Roberta Gottlieb and Grant Meisenholder
• Chapter 5: Measurement of Ligand Acidification Kinetics for Adherant and Non-Adherant Cells; Sheree Lynn Ryback and Robert F. Murphy
• Chapter 6: Detection of Intracellular Antigens by Flow Cytometry; Ingrid Schmid
• Chapter 7: Flow Cytometric Measurement of Nuclear Matrix Proteins; Joe Dynlacht
• Chapter 8: Flow Cytoenzymology using CellProbe Technology; Susan DeMaggio

ELECTRONIC CELL SORTING

• Chapter 1: Creating Standard and Reproducible Sorting Conditions; Marty Bigos, Richard Stoval, and David Parks
• Chapter 2: Sterilization for Sorting; Steve Merlin
• Chapter 3: High Speed Sorting by MoFlo; Peter Lopez
• Chapter 4: Purification of Mouse Fetal Liver Hematopoeitic Stem Cells; Sean Morrison
• Chapter 5: Methods for Preparing Sorted Cells for Further Analysis; Robert Lief and Karen Chew

CORE FACILITIES

• Chapter 1: Core Facilities for Flow Cytometry; Steve Merlin
• Chapter 2: Funding for Core Facilities; Larry Sklar
• Chapter 3: Biosafety in the Flow Cytometry Laboratory; Ingrid Schmid

UNLOCKING THE SPECTRUM OF POSSIBILITIES; Rochelle Diamond

APPENDIX

• Lasers; Paul Ginouves
• Table of Fluorescent Compounds, Applications, Excitation and Emission spectra, Laser Lines, and Filter Setups; Steve Merlin
• Common Filter Setup for Large Sorter System – FACS Vantage; Steve Merlin
• Schematic for following Fluidics Problems; Steve Merlin, Rebecca Poon
• PHK Cell Linker Dyes; Steve Merlin, Rebecca Poon
• Cell Census Software; Rebecca Poon
• Monoclonal Antibody Production; Gary Woo
• Commercial Sources; VARIOUS Authors
• Networking – Electronic and Personal

Glossary