Resch-Genger Standardization and Quality Assurance in Fluorescence Measurements II

Bioanalytical and Biomedical Applications
1. Auflage 2008
ISBN: 978-3-540-70571-0
Verlag: Springer-Verlag
Format: PDF
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)

E-Book, Englisch, 564 Seiten

Analytical chemists and materials scientists will find this a useful addition to their armory. The contributors have sought to highlight the present state of affairs in the validation and quality assurance of fluorescence measurements, as well as the need for future standards. Methods included range from steady-state fluorometry and microfluorometry, microscopy, and micro-array technology, to time-resolved fluorescence and fluorescence depolarization imaging techniques.

Autoren/Hrsg.

Resch-Genger, Ute

Inhaltsverzeichnis

1;Series Editor;6
2;Volume Editor;6
3;Preface;7
4;Contents;9
5;Contributors;12
6;Part I Fluorescence Microscopy;16
6.1;Need for Standardization of Fluorescence Measurements from the InstrumentManufacturer’s View;17
6.1.1;1 Introduction;18
6.1.2;2 Standardization – but Which Parameters?;19
6.1.3;3 Overview on Calibration Methods for Confocal Microscopy;22
6.1.4;4 Use of a Thin Fluorescent Film Sample to Determine the Signal- to- Noise Ratio in a Confocal Microscope;23
6.1.5;5 Conclusion;32
6.1.6;References;38
6.2;Characterization and Calibration inWide Field and Sectioned Fluorescence Microscopy SIPcharts;39
6.2.1;1 Introduction;40
6.2.2;2 Image Calibration in Wide Field Fluorescence Microscopy;41
6.2.3;3 Characterization of Sectioning Fluorescence Microscopy (3D) with Thin Uniform Fluorescent Layers: Sectioned Imaging Property or SIPcharts;55
6.2.4;4 Conclusions;66
6.2.5;References;68
6.3;Quantitative Fluorescence Microscopy: Considerations and Controls;69
6.3.1;1 Introduction;70
6.3.2;2 The Intensity Dimension;72
6.3.3;3 The Spatial Dimension;86
6.3.4;4 The Temporal Dimension;98
6.3.5;5 Discussion;99
6.3.6;References;100
6.4;Comparability of Fluorescence Microscopy Data and Need for Instrument Characterization of Spectral Scanning Microscopes;103
6.4.1;1 Introduction;104
6.4.2;2 Instrument-Specific Parameters and Quantities Affecting Spectral Measurements in Microscopy;107
6.4.3;3 Comparability of Fluorescence Microscopy Data and Instrument Characterization;108
6.4.4;4 Conclusion, Future Requirements and Challenges;126
6.4.5;References;127
6.5;Fluorescence Lifetime Imaging Microscopy: Quality Assessment and Standards;131
6.5.1;1 Introduction;132
6.5.2;2 Förster Resonance Energy Transfer;134
6.5.3;3 Instrumentation and Techniques;135
6.5.4;4 Fluorophore Standards;139
6.5.5;5 System Calibration;141
6.5.6;6 Photon Efficiency and Photon Economy;143
6.5.7;7 Acquisition Speed;145
6.5.8;8 Data Quality Assessment and Analysis;146
6.5.9;9 Photobleaching;149
6.5.10;10 Fluorophore Saturation;150
6.5.11;11 Fluorescent Stainings;151
6.5.12;12 Conclusions;152
6.5.13;References;155
7;Part II Single Molecule Spectroscopy;157
7.1;State of the Art and Novel Trends in Fluorescence Correlation Spectroscopy;158
7.1.1;1 Introduction;159
7.1.2;2 General Principles of FCS;161
7.1.3;3 Experimental Aspects;178
7.1.4;4 Nonidealities and Artifacts;180
7.1.5;5 Evaluation of Experimental FCS Data;187
7.1.6;6 Some Application Aspects;192
7.1.7;7 Varieties of FCS and Related Techniques;197
7.1.8;8 Conclusions;202
7.1.9;References;202
7.2;Single Molecule Spectroscopy: Instrumentation and Multiparameter Detection;211
7.2.1;1 Introduction;211
7.2.2;2 Accessible Parameters;212
7.2.3;3 Instrumentation;215
7.2.4;4 Data Analysis in Multiparameter Measurements;218
7.2.5;References;223
8;Part III Fluorescence- Based Microarray Technology: Applications, Future Trends, and Need for Standardization;225
8.1;DNA Microarrays: Applications, Future Trends, and the Need for Standardization;226
8.1.1;1 Introduction;227
8.1.2;2 Microarray Technology;229
8.1.3;3 Microarray Applications;232
8.1.4;4 Future Trends;235
8.1.5;5 Need for Standardization;240
8.1.6;6 Conclusions;245
8.1.7;References;246
8.2;Comparability of Microarray Experiments from the Instrument and the Sample Site and Approaches Towards Standardization;249
8.2.1;1 Introduction;250
8.2.2;2 General Considerations About Microarray Experiments;251
8.2.3;3 Sources of Inconsistency in Microarray Measurements;257
8.2.4;4 Approaches Towards Standardization;258
8.2.5;5 Comparability of Microarray Experiments;263
8.2.6;6 Commercial DNA Microarray Platforms;267
8.2.7;7 Conclusion;270
8.2.8;References;270
8.3;Microarray Technology: Unresolved Issues and Future Challenges froma Regulatory Perspective;274
8.3.1;1 Introduction: The Apparent Lack of Reproducibility;275
8.3.2;2 Microarray Quality-Control Metrics and Thresholds: Assessing Data Quality from Various Perspectives;276
8.3.3;3 Consensus on Data-Analysis Methods: Adequate Evaluation Is Needed;279
8.3.4;4 The MicroArray Quality Control (MAQC) Project;283
8.3.5;5 Conclusions;288
8.3.6;References;290
8.4;Protein Arrays and Fluorescence Detection: Applications and Limitations;292
8.4.1;1 Introduction;293
8.4.2;2 Peptide and Protein Arrays: Composition, Production, and Processing;294
8.4.3;3 Applications of Protein Arrays;302
8.4.4;4 Quality Assurance of Array Data;308
8.4.5;References;311
9;Part IV Flow Cytometry;314
9.1;Flow Cytometry: Instrumentation, Applications, Future Trends and Limitations;315
9.1.1;1 Introduction;316
9.1.2;2 General Concept of Flow Cytometry with Brief History;317
9.1.3;3 Instrumentation Details;324
9.1.4;4 Optical Measurements;334
9.1.5;5 Limitations;342
9.1.6;6 Applications and Future Trends;345
9.1.7;References;347
9.2;Flow Cytometry Quality Assurance;351
9.2.1;1 Introduction;352
9.2.2;2 Precision;354
9.2.3;3 Sensitivity;363
9.2.4;4 Summary;376
9.2.5;References;376
9.3;Approaches to Quantitation in Flow Cytometry;379
9.3.1;1 Introduction;381
9.3.2;2 Fluorescence Signal;382
9.3.3;3 Conceptual Framework for Quantitation;383
9.3.4;4 Practical Assignment of MESF Values;385
9.3.5;5 Application of MESF;395
9.3.6;6 Conclusion;404
9.3.7;References;405
10;Part V Fluorescence Immunoassays;407
10.1;Immunoassays: Basic Concepts, Physical Chemistry and Validation;408
10.1.1;1 Introduction: Antibodies as an Analytical Tool;409
10.1.2;2 Antibodies and Recognition Reaction;410
10.1.3;3 Assay Formats – Assay Setups;414
10.1.4;4 Marker Systems and Conjugation;421
10.1.5;5 Assay Validation;425
10.1.6;6 Conclusions and Outlook;433
10.1.7;References;434
10.2;Time-Resolved Fluorometric Immunoassays; Instrumentation, Applications, Unresolved Issues and Future Trends;436
10.2.1;1 Immunoassays as Analytical Methods in Diagnostics;436
10.2.2;2 Time-Resolved Fluorometry;438
10.2.3;3 Lanthanide Chelates and Assay Technologies;441
10.2.4;4 Problems of Standardization in Clinical Immunoassays;444
10.2.5;5 Immunofluorometric Applications;446
10.2.6;6 Future Trends and Challenges;449
10.2.7;References;451
10.3;Particle-Based Assays: Applications and Unresolved Issues;455
10.3.1;1 Introduction;456
10.3.2;2 Performance Criteria of Immunoassays;457
10.3.3;3 Why Use Particles?;459
10.3.4;4 General Aspects of Particle-Based Assays;461
10.3.5;5 Issues Concerning Specific Markers;465
10.3.6;6 Conclusion and Outlook;472
10.3.7;References;473
10.4;Advances in Fluorescence Enzyme Detection Methods;475
10.4.1;1 Introduction;476
10.4.2;2 Fluorigenic Substrates – Introduction;478
10.4.3;3 Fluorigenic Substrates Utilising Energy Transfer;482
10.4.4;4 Fluorescence Polarisation Enzyme Assays;484
10.4.5;5 Fluorigenic Substrates Yielding Insoluble Products;487
10.4.6;6 Intracellular Enzyme Assays;488
10.4.7;7 Multiplexed Enzyme Assays;488
10.4.8;8 Conclusions;490
10.4.9;References;490
11;Part VI Quantitative PCR;492
11.1;Quantitative Real-Time PCR: Fluorescent Probe Options and Issues;493
11.1.1;1 PCR;494
11.1.2;2 Real-Time Quantitative PCR (rt-Q-PCR);495
11.1.3;3 Fluorescence Detection in Quantitative PCR: Nonspecific Detection;496
11.1.4;4 Fluorescence Detection in Quantitative PCR: Specific Detection;497
11.1.5;5 Conclusions;510
11.1.6;References;511
12;Part VII Fluorescence In Situ Hybridization and Immunohistochemistry;513
12.1;Cellular Bioimaging in Fluorescent Cancer Biomarker Evaluation: Validation, Technologies and Standards Development;514
12.1.1;1 Introduction: Validation, Heterogeneity and Cellular Cancer Biomarkers;515
12.1.2;2 Continuous and Discrete Values for Cancer Biomarkers, Cellular Heterogeneity and Measurement Issues;518
12.1.3;3 DNA and Antibody Probes: Fluorescence Reagents for Cancer Biomarker Quantification in Cells;520
12.1.4;4 Application to Quantitative Medical Histopathology;527
12.1.5;5 Significant Challenges to Implementation of Standards for Cancer Biomarkers;527
12.1.6;6 Physical Standards for Fluorescent Measurement of Cancer Biomarkers: Now or Later?;528
12.1.7;7 Summary;529
12.1.8;References;530
13;Part VIII Fluorescence Technologies in Biomedical Diagnostics;534
13.1;Fluorescence Techniques in Biomedical Diagnostics: Instrumentation, Analysis and Unresolved Issues;535
13.1.1;1 Overview;536
13.1.2;2 Methods, Instrumentation and Applications;536
13.1.3;3 Conclusions;547
13.1.4;References;548
13.2;In-vivo Fluorescence Imaging: Applications, Future Trends & Approaches to Standardization;551
13.2.1;1 Main Text;551
13.2.2;2 Diffuse Fluorescence Tomography;553
13.2.3;3 Relation of Inversion and Standardization;555
13.2.4;4 Current Diffusive Fluorescent Standards;558
13.2.5;5 Conclusion;561
13.2.6;References;561
14;Subject Index;563