FLIM Microscopy in Biology and Medicine

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Ammasi Periasamy, Robert M. Clegg
CRC Press, Jul 6, 2009 - Science - 472 pages
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Detecting Signals at the Single Molecule Level: Pioneering Achievements in Microscopy

Recent advances have led to such remarkable improvements in fluorescence lifetime imaging microscopy’s (FLIM) capacity for contrast and sensitivity that researchers can now employ it to detect signals at the single molecule level. FLIM also offers the additional benefit of independence from fluorophore concentration and excitation intensity. Moreover, its unique sensitivity makes it an excellent reporter of conformational changes and of variations in the molecular surroundings of biological molecules.

Most of this improvement and discovery have occurred during the past decade, and, to date, information that would benefit a broad range of researchers remains scattered in the literature. Edited by two of the top pioneers in the field, FLIM Microscopy in Biology and Medicine presents the fundamentals of FLIM along with a number of advanced considerations so that a wider audience can appreciate recent and potential improvements that make it such a valuable tool.

New Opportunities for Biomedical Researchers... New Challenges for Microscopy Researchers

Discussion sections in all the chapters clearly show the challenges for implementing FLIM for various applications. Certain chapters discuss limits on the number of photons required for highly accurate lifetime determinations, as well as the accuracy with which multiple, closely associated lifetime components can reliably be determined. Such considerations are important for the user when he or she is selecting the most advantageous method of FLIM to use for a particular application.

While this book provides an introduction for those new to FLIM, it gathers a wealth of material to enhance the work of experts involved in pioneering technological improvements, as well as those research opportunities in this unique and promising area of microscopy.

 

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I found this book great for this vast field of analysis between theoretical computer scientists and discrete mathematicians. Lots of great material and great coverage across the board. I use this for my research.

Contents

The Editors
xxv
Contributors
xxvii
Introduction Microscopy Fluorophores
1
What Why HowA Prologue
3
Chapter 2 Principles of Fluorescence for Quantitative Fluorescence Microscopy
35
Chapter 3 Visible Fluorescent Proteins for FRETFLIM
65
Instrumentation
91
Chapter 4 WideField Fluorescence Lifetime Imaging Mircoscopy Using a Gated Image Intensifier Camera
93
Chapter 9 Spectrally Resolved Fluorescence Lifetime Imaging Microscopy
211
Chapter 10 TimeResloved Fluorescence Anisotropy
245
Data Analysis
289
FrequencyDomain ModelFree Analysis
291
Chapter 12 Nonlinear CurveFitting Methods for TimeResolved Data Analysis
341
Chapter 13 Global Analysis of Frequency Domain FLIM Data
371
Applications
383
Chapter 14 FLIM Applications in the Biomedical Sciences
385

Chapter 5 FrequencyDomain FILM
115
Chapter 6 Laser Scanning Confocal FLIM Microscopy
143
Chapter 7 Multiphoton Fluorescence Lifetime Imaging at the Dawn of Clinical Application
165
Monitoring Metabolic Processes in Living Cells and Tissues
189
Index
401
Back cover
441
Copyright

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About the author (2009)

Ammasi Periasamy, University of Virginia, Charlottesville, USA

Robert M. Clegg, University of Illinois, Urbana-Champaign, USA

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