Methods in Cell Biology: Flow Cytometry Vol 42
Zbigniew Darzynkiewicz
J.Paul Robinson
Harry A. Crissman

Academic Press, San Diego, 1994
ISBN 0-12-203052-4 [Softcover]
ISBN 0-12-564143-5 [Hardcvover]


CONTENTS

   

Contents of Volume 42

xvii

Contributors

xxi

Preface to the Second Edition

xxvii

Preface to the First Edition

xxxi

   

1. A Flow Cytometric Technique for Detection of DNA Repair in Mammalian Cells

 

Jules R. Selden and Frank Dolbeare

 
   

I. Introduction

1

II. Application

3

III. Materials

4

IV. Sample Preparation and Staining

5

V. Critical Aspects ofthe Procedure

9

VI. Controls and Standards

11

VII. Instruments and Setup for Data Collection

12

VIII. Results and Discussion

13

References

18

   

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

 

Awtar Krishan

 
   

I. Introduction

21

II. Application

22

III. Materials

23

IV. Staining Procedure

24

V. Critical Aspects

24

VI. Controls and Standards

25

VII. Instruments

25

VIII. Results

26

References

30

   

3. Glutathione and Cellular Resistance to Anti-Cancer Drugs

 

David Hedley and Sue Chow

 
   

I. Introduction

31

II. Flow Cytometric Glutathione Probes

33

III. Materials

36

IV. Methods

37

References

44

   

4. Assay of Cell Resistance to Ara-C

 

Francis Lacombe

 
   

I. Introduction

45

II. Application

47

III. Materials

48

IV. Cell Preparation and Staining

48

V. Critical Aspects of the Procedure

49

VI. Instruments

49

VII. Results and Discussion

52

VIII. Comparison of the Methods

56

References

57

   

5. Detection of mRNA Species by Flow Cytometry

 

Francis Belloc and Francoise Durrieu

 
   

I. Introduction

59

II. Applications

61

III. Materials

62

IV. Staining Procedures

63

V. Critical Aspects of the Procedure and Controls

64

VI. Instruments and FCM Analysis

66

VII. Results and Discussion

67

References

68

   

6. Primed in Situ Labeling (PRINS) and Fluorescence in Situ Hybridization (FISH)

 

Hanne Fischer, Johnny Hindkjoer, Soren Pedersen, Jorn Koch, Carsten Brandt, and Steen K0luroa

 
   

I. Introduction

72

II. Applications

72

III. Materials

73

IV. Protocols

76

V. Critical Aspects of the Procedures

87

VI. Instruments

89

VII. Results and Discussion

90

VIII. Comparison of Methods

92

References

93

   

7. Molecular Phenotyping by Flow Cytometry

 

Benjamin D. Li, Earl A. Timm, Jr., Mary C. Riedy, Seth P. Harlow, and Carleton C. Stewart

 
   

I. Introduction

96

II. General Materials and Methods

97

III. Measurement of Gene Amplification Using Competitive PCR

101

IV. Measurement of Gene Expression Using Competitive RT-PCR

115

V. Fluorescence in Situ Hybridization En Suspension (FISHES)

121

VI. Fluorescence in Situ PCR (FLIP)

126

References

129

   

8. Isolation and Analysis of Somatic Cell Mutants with Defects in Endocytic Traffic

 

Sandra A. Brockman and Robert F. Murphy

 
   

I. Introduction: Why Isolate Somatic Cell Mutants in Membrane Traffic?

131

II. General Considerations

133

III. Isolation of CHO Cells Defective in Receptor Recycling

139

IV. Isolation of B Cells with Reduced Cysteine Protease Activity 142

 

V. Conclusions

146

References

147

   

9. Measurement of Micronuclei by Flow Cytometry

 

Michael Nusse, Wolfgang Beisker, Johannes Kramer, Beate M. Miller, Georg A. Schreiber, Silvia Viaggi, Eva Maria Weller, and Jurina M. Wessels

 
   

I. Introduction

149

II. Application

150

III. Materials

151

IV. Cell Preparation and Staining

152

V. Critical Aspects ofthe Procedure

152

VI. Standards

153

VII. Instruments

154

VIII. Results and Discussion

155

References

158

   

10. Sperm Chromatin Structure Assay: DNA Denaturability

 

Donald Evenson and Lorna Jost

 
   

I. Introduction

160

II. Applications ofthe SCSA

161

III. Materials

163

IV. Cell Preparation

164

V. Cell Staining and Measurement

166

VI. Instruments

168

VII. Results and Discussion

169

VIII. Critical Points

173

References

1 75

   

11. Fine-Needle Cytopuncture and Flow Cytometry in the Characterization of Human Tumors

 

Frederique Spyratos and Marianne Brifford

 
   

I. Introduction

178

II. Materials and Methods

178

III. Critical Aspects

181

IV. Alternative Methods Applied to Cytopunctures

182

V. Clinical Applications

187

VI. Conclusion

191

References

192

Note Added in Proof

192

   

12. Functional NK Assays Using Flow Cytometry

 

Stefano Papa and Massimo Valentini

 
   

I. Introduction

194

II. Application

196

III. Materials

196

IV. Cell Preparation and Staining

197

V. Critical Aspects ofthe Procedures

198

VI. Standardization

200

VII. Instruments

200

VIII. Results and Discussion

201

References

206

   

13. Antibodies to Intermediate Filament Proteins as Probes for Multiparameter Flow Cytometry of Human Cancers

 

T. Vincent Shankey

 
   

I. Introduction

209

II. Intermediate Filament Proteins

210

III. Cell Isolation and Fixation

214

IV. Multiparameter Flow Cytometric Analysis Using Antibodies

216

to Cytokeratins

 

V. Clinical Application Using Multiparameter DNA Content

222

Flow Cytometry

 

VI. Conclusions

227

References

228

   

14. Interactive Data Analysis for Evaluation of B-Cell Neoplasia by Flow Cytometry

 

Ricardo E. Duque

 
   

I. Background

231

II. Correlated Multiparametric Data Analysis of B-Cell Neoplasia

235

References

241

   

15. Analysis and Sorting of Hematopoietic Stem Cells from Mouse Bone Marrow

 

Jan W. M. Visser and Peter de Vries

 
   

I. Introduction

244

II. Application

247

III. Materials

248

IV. Instruments

250

V. Staining and Sorting Procedure

250

VI. Critical Aspects of the Procedure

255

VII. Results and Discussion

257

References

260

   

16. Reticulocyte Analysis and Reticulocyte Maturity Index

 

Bruce H. Davis and Nancy C. Bigelow

 
   

I. Introduction

263

II. Reticulocyte Maturity Index

266

III. Data Analysis

267

IV. Quality Control

270

V. Method for Thiazole Orange Reticulocyte Analysis

271

References

273

   

17. Analysis of Platelets by Flow Cytometry

 

Kenneth A. Ault and fane Mitchell

 
   

I. Introduction

275

II. Applications

277

III. Materials

279

IV. Cell Preparation

280

V. Staining

283

VI. Critical Aspects

285

VII. Standards

286

VIII. Instrument

287

IX. Results and Discussion

287

References

293

   

18. Flow Cytometry in Malaria Detection

 

Chris J. Fanse and Philip H. Van Vianen

 
   

I. Introduction

296

II. Applications

297

III. Materials

300

IV. Cell Preparation and Staining

300

V. Critical Aspects of the Preparation and Staining Procedures

304

VI. Standards

305

VII. Instruments

305

   

VIII. Results and Discussion

307

IX. Comparison of Methods

316

References

318

   

19. Large-Scale Chromosome Sorting

 

John J. Famcett, Jonathan L. Longmire, John C. Martin, Larry L. Deaven, and L. Scott Cram

 
   

I. Introduction

319

II. Sample Preparation

320

III. Preparation Evaluation

324

IV. Production Chromosome Sorting

325

Appendix: Polyamine Sheath Buffer

328

References 329

 
   

20. Strategies for Rare Cell Detection and Isolation

 

James F. Leary

 
   

I. Introduction

332

II. Methods

341

III. Critical Aspects of the Procedure

348

IV. Instrumentation

354

V. Results: Some Rare Cell Applications and Strategies

356

VI. Discussion

357

References

358

   

21. Cell Sorting of Biohazardous Specimens for Assay of Immune Function

 

Janis V. Giorgi

 
   

I. Introduction

359

II. Applications

360

III. Critical Aspects of the Procedure

360

IV. Instruments

362

V. Verification of Aerosol Containment

363

VI. Staining and Cell Separation of Potentially Biohazardous Specimens

367

VII. Results and Discussion

368

VIII. Alternate Technologies for Cell Separation

368

References

369

   

22. High-Speed Photodamage Cell Sorting: An Evaluation of the

 

ZAPPER Prototype

 

Jan F. Keij, Ad C. Groenewegen, andJan W. M. Visser

 
   

I. Introduction

372

II. Application

373

III. Materials

373

IV. Critical Aspects of the Procedure

377

V. Instrumentation

381

VI. Results and Discussion

382

References

385

   

23. High-Gradient Magnetic Cell Sorting

 

Andreas Radbruch, Birgit Mechtold, Andreas Thiel, Stefan Miltenyi, and Eckhard Pfluger

 
   

I. Introduction

388

II. Application

388

III. Materials

390

IV. MACS: Staining and Sorting

391

V. Critical Aspects of the Procedure

395

VI. Controls

397

VII Instruments

397

VIII. Results

399

References

402

   

24. Contributions of Flow Cytometry to Studies with Multicell Spheroids

 

Ralph E. Durand

 
   

I. Introduction

406

II. Spheroid Growth and Analysis

406

III. Flow Cytometry Techniques

407

IV. Studies of Spheroid Oxygenation

408

V. Evaluation of Antineoplastic Drugs

410

VI. Drug Interactions: Synergy in Cell Subpopulations

414

VII. Multifraction Treatments

415

VIII. Cell Kinetics: Role of Pretreatment Kinetics

417

IX. Alternatives for Kinetic Measurements: Overcoming Heterogeneity

419

X. General Conclusions

420

References

421

   

25. Functional Measurements Using HL-60 Cells

 

J. Paul Robinson, Padma Kumar Narayanan, and Wayne 0. Carter

 
   

I. Introduction

423

II. Application

424

III. Materials and Methods

425

IV. Results and Discussion

431

References

436

   

26. HIV Infection: Diagnosis and Disease Progression Evaluation

 

Janis V. Giorgi and Alan Landay

 
   

I. Introduction

438

II. Application

439

III. Materials

439

IV. Cell Preparation and Staining

442

V. Critical Aspects of the Procedure

445

VI. Controls and Standards

450

VII. Instruments

452

VIII. Results and Discussion

452

IX. Alternative Technologies

453

References

454

   

27. Cell-Cycle Analysis of Saccharomyces cerevisiae

 

Bruce S. Dien, Marvin S. Peterson, and Friedrich Srienc

 
   

I. Introduction

458

II. Materials

460

III. Methods

461

IV. Critical Aspects

463

V. Flow Cytometry

465

VI. Analysis of DNA Histograms

466

VII. Results

466

References

474

   

28. Staining and Measurement of DNA in Bacteria

 

Harald B. Steen, Mette W. Jernaes, Kirsten Skarstad, and Erik Boye

 
   

I. Introduction

477

II. Materials

480

III. Fixation and Staining

481

IV. Vital Staining

481

V. Flow Cytometric Measurement

 

VI. Standards and Controls

484

References

487

   

29. Detection of Specific Microorganisms in Environmental Samples Using Flow Cytometry

 

Graham Vesey, foe Narai, Nicholas Ashbolt, Keith Williams, and Duncan Veal

 
   

I. Introduction

490

II. Preparation of Water Samples for Flow Cytometric Analysis

499

III. Staining of Organisms from Water Samples for Flow

501

Cytometric Analysis

 

IV. Flow Cytometric Analysis of Water Samples

509

V. Instrumentation Developments for Environmental Applications

516

VI. The Future of Flow Cytometry within Environmental Microbiology

518

References

519

   

30. Strategies for Flow Cytometric Analysis of Marine Microalgae and Sponge Cells

 

Clarice M. Yentsch and Shirley A. Pomponi

 
   

I. Introduction

524

II. Application

524

III. Materials

526

IV. Cell Preparation and Staining

526

V. Instrumentation

532

VI. Results

536

VII. Discussion

537

References

538

   

31. Flow Cytometry and Sorting of Plant Protoplasts and Cells

 

David W. Galbraith

 
   

I. Introduction

540

II. Application

542

III. Materials

542

IV. Procedures

543

V. Critical Aspects of the Procedures

550

VI. Controls and Standards

555

VII. Instruments

555

VIII. Results

556

References

560

   

32. Use of Fluorescence-Activated Cell Sorting for Rapid Isolation of Insect Cells Harboring Recombinant Baculovirus

 

Maja A. Sommerfelt and Eric J. Sorscher

 
   

I. Introduction

564

II. Application

565

III. Materials

565

IV. Methods

566

V. Critical Aspects of the Procedure

570

VI. Instruments

570

VII. Results and Discussion

570

VIII. Comparison of Methods

572

References

573

   

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

 

Thomas M. McHugh

 
   

I. Introduction

575

II. Instrumentation and Materials

578

III. Procedures

580

IV. Review of Published Applications

590

V. Summary

594

VI. Suppliers

594

References

595

   

34. Calibration of Flow Cytometer Detector Systems

 

Ralph E. Durand

 
   

I. Introduction

597

II. Materials

598

III. Procedures

599

IV. Concluding Comments

603

References

604

   

35. Standardization for Flow Cytometry

 

A. Schwartz and Emma Fernandez-Repollet

 
   

I. Introduction

605

II. Considerations for Qualitative Standardization

606

III. Considerations for Quantitative Standardization

619

IV. Conclusions

624

References

625

   

36. Phase-Sensitive Detection Methods for Resolving Fluorescence Emission Signals and Directly Quantifying Lifetime

 

John A. Steinkamp

 
   

I. Introduction

627

II. Application

632

III. Cell Preparation and Staining

633

IV. Critical Aspects ofthe Procedure

633

V. Standards

635

VI. Instrumentation

635

VII. Results and Discussion

636

References

640

   

37. Spectra of Fluorescent Dyes Used in Flow Cytometry

 

Richard P. Haugland

 
   

I. Introduction

641

II. Spectral Properties of Fluorescent Dyes

642

III. Immunofluorescence and Related Applications

645

IV. Nucleic Acid Stains

652

V. Probes for Live Cell Physiology

655

VI. Summary

661

References

662

   

Index

665

Volumes in Series

693