CONTENTS

CONTRIBUTORS

xi

PREFACE

XV

1. Dissociation of Cells from Solid Tumors

1

Robert Cerra, Richard J. Zarbo, and John D. Crissman

2. Flow Cytometric Measurement of Cell Viability Using DNase Exclusion

13

Oskar S. Frankfurt

3. Cell Cycle Phase-Specific Analysis of Cell Viability Using Hoechst 33342 and Propidium Iodide after Ethanol Preservation

19

Alan Pollack and Gaetano Ciancio

4. Cell Membrane Potential Analysis

25

Howard M. Shapiro

5. Flow Cytometric Measurement of Intracellular Ionized Calcium in Single Cells with Indo-1 and Fluo-3

37

Carl H. June and Peter S. Rabinovitch

6. Measurement of Intracellular pH

59

Elizabeth A. Musgrove and David W. Hedley

7. Flow Cytometric Techniques for Measurement of Cytochrome P-450 Activity in Viable Cells

71

A. Dusty Miller

8. Fluorescent Staining of Enzymes for Flow Cytometry

81

Frank Dolbeare

9. Supravital Cell Staining with Hoechst 33342 and DiOC563J

89

Harry A. Crissman, Marianne H. Hofland, Anita P. Stevenson, Mark E. Wilder, and Robert A. Tobey

10. Specific Staining of DNA with the Fluorescent Antibiotics, Mithramycin, Chromomycin, and Olivomycin

97

Harry A. Crissman and Robert A. Tobey

11. DAPI Staining of Fixed Cells for High-Resolution Flow Cytometry of Nuclear DNA

105

Friedrich Otto

12. High-Resolution DNA Measurements Using the Nuclear Isolation Medium, DAPI, with the RATCOM Flow Cytometer

111

Jerry T. Thornthwaite and Richard A. Thomas

13. Rapid DNA Content Analysis by the Propidium Iodide-Hypotonic Citrate Method

121

Awtar Krishan

14. An Integrated Set of Methods for Routine Flow Cytometric DNA Analysis

127

Lars Vindelov and Ib Jarle Christensen

15. DNA Analysis from Paraff n-Embedded Blocks

139

David W. Hedley

16. Detection of M and Early-G1 Phase Cells by Scattering Signals Combined with Identifcation of G1, S, and G2 Phase Cells

149

Elio Geido, Walter Giaretti, and Michael Nusse

17. Controls, Standards, and Histogram Interpretation in DNA Flow Cytometry

157

Lynn G. Dressler

18. Cell Division Analysis Using Bromodeoxyuridine-Induced Suppression of Hoechst 33258 Fluorescence

173

Ralph M. Bohmer

19. Cell Cycle Analysis Using Continuous Bromodeoxyuridine Labeling and Hoechst 33258-Ethidium Bromide Bivariate Flow Cytometry

185

Martin Poot, Manfred Kubbies, Holger Hoehn, Angelika Grossmann, Yuhchyou Chen, and Peter S. Rabinovitch

20. Detection of Bromodeoxyuridine-Labeled Cells by Differential Fluorescence Analysis of DNA Fluorochromes

199

Harry A. Crissman and John A. Steinkamp

21. Using Monoclonal Antibodies in Bromodeoxyuridine-DNA Analysis

207

Frank Dolbeare, Wen-Lin Kuo, Wolfgang Beisker, Martin Vanderlaun, and Joe W. Gray

22. Identification of Proliferating Cells by Ki-67 Antibody

217

Heinz Baisch and Johannes Gerdes

23. Washless Double Staining of a Nuclear Antigen (Ki-67 or Bromodeoxyuridine) and DNA in Unfixed Nuclei

227

Jorgen K. Larsen

24. Analysis of Proliferation-Associated Antigens

235

Kenneth D. Bauer

25. The Stathmokinetic Experiment: A Single-Parameter and Multiparameter Flow Cytometric Analysis

249

Frank Traganos and Marek Kimmel

26. Detection of Intracellular Virus and Viral Products

271

Judith Laffin and John M. Lehman

27. Differential Staining of DNA and RNA in Intact Cells and Isolated Cell Nuclei with Acridine Orange

285

Zbigniew Darzynkiewicz

28. Flow Cytometric Analysis of Double-Stranded RNA Content Distributions

299

Oskar S. Frankfurt

29. Simultaneous Fluorescent Labeling of DNA, RNA, and Protein 305

Harry A. Crissman, Zbigniew Darzynkiewicz, John A. Steinkamp, and Robert A. Tobey

30. Flow Cytometric Cell-Kinetic Analysis by Simultaneously Staining Nuclei with Propidium Iodide and Fluorescein Isothiocyanate

315

Alan Pollack

31. Flow Cytometric Methods for Studying Isolated Nuclei: DNA Accessibility to DNase I and Protein-DNA Content

325

Ryuji Higashikubo, William D. Wright, and Joseph L. Roti Roti

32. Acid-Induced Denaturation of DNA in Situ as a Probe of Chromatin Structure

337

Zbigniew Darzynkiewicz

33. Fluorescent Methods for Studying Subnuclear Particles

353

William D. Wright, Ryuji Higashikubo, and Joseph L. Roti Roti

34. Chromosome and Nuclei Isolation with the MgSO4 Procedure

363

Barbara Trask and Ger van den Engh

35. Univariate Analysis of Metaphase Chromosomes Using the Hypotonic Potassium Chloride-Propidium Iodide Protocol

369

L. Scott Cram, Frank A. Ray, and Marty F. Bartholdi

36. Polyamine Buffer for Bivariate Human Flow Cytogenetic Analysis and Sorting

377

L. Scott Cram, Mary Campbell, John J. Fawcett, and Larry L. Deaven

37. Fluorescence in Situ Hybridization with DNA Probes

383

Barbara Trask and Dan Pinkel

38. Flow Cytometric Analysis of Male Germ Cell Quality

401

Donald P Evenson

39. Cell Preparation for the Identification of Leukocytes

411

Carleton C. Stewart

40. Multiparameter Analysis of Leukocytes by Flow Cytometry

427

Carleton C. Stewart

41. Identification and Purification of Murine Hematopoietic Stem Cells by Flow Cytometry

451

Jan W. M. Visser and Peter de Vries

42. Fluorescent Cell Labeling for in vivo and in Vitro Cell Tracking

469

Paul Karl Horan, Meryle ~ Melnicoff, Bruce D. Jensen, and Sue E. Slezak

43. Rapid Determination of Cellular Resistance-Related Drug Efflux in Tumor Cells

491

Awtar Krishan

44. Slit-Scan Flow Analysis of Cytologic Specimens from the Female Genital Tract

501

Leon L. Wheeless, Jr., Jay E. Reeder, and Mary J. O'Connell

45. Cell Sorting with Hoeschst or Carbocyanine Dyes as Perfusion Probes in Spheroids and Tumors

509

Ralph E. Durand, David J. Chaplin, and Peggy L. Olive

46. DNA Measurements of Bacteria

519

Harald B. Steen, Kirsten Skarstad, and Erik Boye

47. Isolation and Flow Cytometric Characterization of Plant Protoplasts

527

David W. Galbraith

48. Flow Cytometric Analysis of Plant Genomes

549

David W. Galbraith

49. Plant Cell Cycle Analysis with Isolated Nuclei

563

Catherine Bergounioux and Spencer G Brown

50. Environmental Health Flow Cytometric Methods to Assess Our Water World

575

Clarice M. Yentsch

51. Flow Microsphere Immunoassay for the Quantitative and Simultaneous Detection of Multiple Soluble Analytes

613

Mack J Fulwyler and Thomas M. McHugh

52. On-Line Flow Cytometry: A Versatile Method for Kinetic Measurements

631

Kees Nooter, Hans Herweijer, Richard Jonker, and Ger van den Engh

53. Calibration of Flow Cytometer Detector Systems

647

Ralph E. Durand

54. Spectral Properties of Fluorochromes Used in Flow Cytometry

655

Jan Kapuscinski and Zbigniew Darzynkiewicz

INDEX

671

CONTENTS OF RECENT VOLUMES

711