Fluorescence-Lifetime Imaging Techniques for Microscopy

CHAPTER 21 Fluorescence-Lifetime Imaging Techniques for Microscopy Chen Y. Dong, 1 Todd French, 2 Peter T. C. So, 1 C. Buehler, 1 Keith M. Berland, 3 and Enrico Gratton 1 Laboratory for Fluorescence Dynamics Department of Physics University of Illinois at Urbana-Champaign Urbana, Illinois 61801 Molecular Devices Corporation Sunnyvale, California 94089 Physics Department Emory University Atlanta, Georgia 30322 I. Introduction II. Time-Resolved Fluorescence Methods A. Time-Domain and Frequency-Domain Measurements B. Simultaneous Multiple-Lifetime-Component Measurement C. Photobleaching Effects III. Fluorescence-Lifetime-Resolved Camera A. Instrumentation B. Camera-Based Microscope Examples IV. Two-Photon Fluorescence Lifetime Microscopy A. Instrumentation B. Two-Photon Microscopy Examples V. Pump-Probe Microscopy A. Instrumentation B. Pump-Probe Microscopy Examples VI. Conclusion References METHODS IN CELL BIOLOGY, VOL. 72 Copyright 1998, Elsevier Inc. All rights reserved. 0091-679X/03 $35.00