Discusses the basic physical principles underlying Biomedical Photonics, spectroscopy and microscopy This volume discusses biomedical photonics, spectroscopy and microscopy, the basic physical principles underlying the technology and its applications.

List of Contributors ix Preface xiii 1 Fluorescence 1 David J. S. Birch, Yu Chen, and Olaf J. Rolinski 1.1 Introduction 1 1.2 Spectra 2 1.3 Quantum Yield 6 1.4 Lifetime 12 1.5 Quenching 23 1.6 Anisotropy 30 1.7 Microscopy 38 1.8 Conclusions 48 Acknowledgments 48 References 49 2 Single-Molecule Detection and Spectroscopy 59 Michel Orrit 2.1 Introduction 59 2.2 Experimental Setups 62 2.3 Fluorescence Spectroscopy 66 2.4 Fluorescence Correlation Spectroscopy 72 2.5 Fluorescence Excitation Spectroscopy 78 2.6 Other Detection Methods 86 2.7 Conclusion 93 Acknowledgments 94 References 94 3 Resonance Energy Transfer 101 David L. Andrews, David S. Bradshaw, Rayomond Dinshaw, and Gregory D. Scholes 3.1 Introduction 101 3.2 History of RET 102 3.3 The Photophysics of RET 103 3.4 Investigative Applications of RET in Molecular Biology 113 3.5 The Role of RET in Light-Harvesting Complexes 118 Acknowledgments 122 References 122 4 Biophotonics of Photosynthesis 129 Valter Zazubovich and Ryszard Jankowiak 4.1 Introduction 129 4.2 Structure of Pigment–Protein Complexes and Structure–Function Relationships 130 4.3 Key Concepts in Physics of Pigment–Protein Complexes 133 4.4 Experimental Techniques 141 4.5 Examples 145 4.6 Conclusions 156 Acknowledgments 157 References 157 5 Optical Sectioning Microscopy and Biological Imaging 165 John Girkin 5.1 Introduction and Background 165 5.2 Confocal Imaging 168 5.3 Nonlinear Microscopy 172 5.4 Practical Implementation of Nonlinear Microscopy 181 5.5 Recent Advances in Nonlinear Microscopy 184 5.6 Widefield Optical Sectioning by Specialized Illumination Methods 186 5.7 Summary 190 References 191 6 Cell Handling, Sorting, and Viability 197 Darwin Palima, Thomas Aabo, Andrew Bañas, and Jesper Glückstad 6.1 Handling Cells with Light 198 6.2 Optical Sorting 215 6.3 Cell Viability 220 References 230 7 Tissue Polarimetry 239 Alex Vitkin, Nirmalya Ghosh, and Antonello de Martino 7.1 Introduction 239 7.2 Polarized Light Fundamentals 240 7.3 Instrumentation 266 7.4 Forward Modeling and Testing in Phantoms 282 7.5 Applications 297 7.6 Conclusions and Outlook 313 References 314 8 Optical Waveguide Biosensors 323 Daphné Duval and Laura M. Lechuga 8.1 Introduction 323 8.2 Fundamentals of Label-Free Optical Waveguide Biosensing 324 8.3 Surface Biofunctionalization 328 8.4 Evaluation of Optical Biosensors 331 8.5 Integrated Optical Waveguide-Based Biosensors 334 8.6 Optical Fiber-Based Biosensors 349 8.7 Lab-On-A-Chip Integration 354 8.8 Summary 357 References 358 9 Light Propagation in Highly Scattering Turbid Media: Concepts, Techniques, and Biomedical Applications 367 R. R. Alfano, W. B. Wang, L. Wang, and S. K. Gayen 9.1 Introduction 367 9.2 Physics Behind Optical Imaging Through a Highly Scattering Turbid Medium 369 9.3 Study of Ballistic and Diffused Light Components 378 9.4 Photon-Sorting Gates 384 9.5 Transition From Ballistic to Diffuse Imaging in Turbid Media 402 9.6 Conclusion 404 Acknowledgments 404 References 404 10 Photodynamic Therapy 413 Rakkiyappan Chandran, Tyler G. St. Denis, Daniela Vecchio, Pinar Avci, Magesh Sadasivam, and Michael R. Hamblin 10.1 Historical Overview of PDT 413 10.2 Introduction to PDT 415 10.3 Photosensitizer Structure and Photophysical Properties 418 10.4 Light Dosimetry and Photodynamic Therapy Light Sources 422 10.5 Light-Based Strategies to Enhance PDT 423 10.6 PDT Targeting and Nanotechnology 425 10.7 PDT for Dermatology 428 10.8 PDT for Oncology 433 10.9 PDT for Infectious Disease 435 10.10 PDT in Ophthalmology 445 10.11 PDT and The Immune System 446 10.12 Conclusion 449 Acknowledgment 449 References 449 11 Imaging and Probing Cells Beyond the Optical Diffraction Limit 469 Mark Schüttpelz and Thomas Huser 11.1 The Quest for Achieving Optical Resolution Beyond ABBE’S Limit 469 11.2 Stimulated Emission Depletion Microscopy 474 11.3 Photoactivated Localization Microscopy and Stochastic Optical Reconstruction Microscopy 477 11.4 Structured Illumination Microscopy 483 11.5 Super-Resolution Optical Fluctuation Imaging and Other Approaches 491 11.6 Outlook 495 Acknowledgments 496 References 497 12 Technology 503 Ann E. Elsner and Christopher A. Clark 12.1 Basic Ocular Anatomy and Physiology 503 12.2 Measurement Techniques 514 12.3 Anterior Segment Diagnostics, Refractive Measurements, and Treatment 522 12.4 Diagnostic Applications and Treatment of Posterior Segment 529 References 534 Index 543

Discusses the basic physical principles underlying biomedical photonics, spectroscopy, and microscopy This volume discusses biomedical photonics, spectroscopy, and microscopy, the basic physical principles underlying the technology and its applications. The topics discussed in this volume are: Fluorescence; Single Molecule Detection and Spectroscopy; Resonance Energy Transfer; Biophotonics of Photosynthesis; Optical Sectioning Microscopy and Biological Imaging; Cell Handling and Sorting; Tissue Polarimetry; Optical Waveguide Biosensors; Light Propagation in Highly Scattering Turbid Media; Photodynamic Therapy; Imaging and Probing Cells beyond the Diffraction Limit; and Ophthalmic Technology. Comprehensive and accessible coverage of the whole of modern photonicsEmphasizes processes and applications that specifically exploit photon attributes of lightDeals with the rapidly advancing area of modern opticsChapters are written by top scientists in their field Written for the graduate-level student in physical sciences; industrial and academic researchers in photonics, graduate students in the area; college lecturers, educators, policymakers, consultants, scientific and technical libraries, government laboratories, NIH.