Principles of Fluorescence Spectroscopy

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752 FLUORESCENCE-LIFETIME IMAGING MICROSCOPY 22.7. CONCLUSIONS FLIM is a robust method for molecular imaging that can be accomplished using several approaches, some of which are listed in the section entitled Additional Reading on Fluorescence-Lifetime Imaging Microscopy near the end of this chapter. FLIM can be used for imaging of ions, biomolecule proximity, and conditions that result in a change in the lifetime of the fluorophore. The instrumentation of FLIM is becoming simpler and less expensive, and can be created using modest modifications of existing instruments. The use of FLIM and the range of its applications is likely to expand greatly with the near future. REFERENCES 1. Lakowicz JR, Berndt KW. 1991. Lifetime-selective fluorescence imaging using an rf phase-sensitive camera. Rev Sci Instrum 62(7): 1727–1734. 2. Lakowicz JR, Szmacinski H, Nowaczyk K, Berndt BW, Johnson ML. 1992. Fluorescence lifetime imaging. Anal Biochem 202:316–330. 3. Szmacinski H, Lakowicz JR, Johnson ML. 1994. Fluorescence lifetime imaging microscopy: homodyne technique using high-speed gated image intensifier. Methods Enzymol 240:723–748. 4. Lakowicz JR, Szmacinski H, Nowaczyk K, Berndt KW, Johnson ML. 1993. Fluorescence lifetime imaging and application to Ca 2+ imaging. In Fluorescence spectroscopy: new methods and applications, chap. 10. Ed OS Wolfbeis. Springer-Verlag, New York. 5. Veselova TV, Cherkasov AS, Shirokov VI. 1970. Fluorometric method for individual recording of spectra in systems containing two types of luminescent centers. Opt Spectrosc 29:617–618. 6. Lakowicz JR, Szmacinski H, Nowaczyk K, Johnson ML, 1992. Fluorescence lifetime imaging of calcium using Quin-2. Cell Calcium 13:131–147. 7. Lakowicz JR, Szmacinski H, Nowaczyk K, Johnson ML. 1994. Fluorescence lifetime imaging of intracellular calcium in COS cells using Quin-2. Cell Calcium 15:7–27. 8. Gerritsen H, Draaijer A, Lakowicz JR. 1997. Introduction, second international lifetime imaging meeting, Utrecht, the Netherlands, June 14, 1996. J Fluoresc 7(1):1–98. 9. Draaijer A, Sanders R, Gerritsen HC. 1995. Fluorescence lifetime imaging, a new tool in confocal microscopy. In Handbook of biological confocal microscopy, pp. 491–505. Ed JB Pawley. Plenum Press, New York. 10. Herman B, Wang XF, Wodnicki P, Perisamy A, Mahajan N, Berry G, Gordon G. 1999. Fluorescence lifetime imaging microscopy. In Applied fluorescence in chemistry, biology and medicine, pp. 491–507. Ed W Rettig, B Strehmel, S Schrader, H Seifert. Springer, Berlin. 11. Clegg RM, Holub O, Gohlke C. 2003. Fluorescence lifetimeresolved imaging: measuring lifetimes in an image. Methods Enzymol 360:509–542. 12. Centonze VE, Sun M, Masuda A, Gerritsen H, Herman B. 2003. Methods Enzymol 360:542–560. 13. Chen Y, Mills JD, Periasamy A. 2003. Protein localization in living cells and tissues using FRET and FLIM. Differentiation 71:528–541. 14. Gadella TWJ, Jovin TM, Clegg RM. 1993. Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond time scale. Biophys Chem 48:221–239. 15. Gadella TWJ, van Hoek A, Visser AJWG. 1997. Construction and characterization of a frequency-domain fluorescence lifetime imaging microscopy system. J Fluoresc 7(1):35–43. 16. Birmingham JJ. 1997. Frequency-domain lifetime imaging methods at unilever research. J Fluoresc 7(1):45–54. 17. Squire A, Verveer PJ, Bastiaens PIH. 2000. Multiple frequency fluorescence lifetime imaging microscopy. J Microsc 197(pt.2):136–149. 18. Clayton AHA, Hanley QS, Arndt-Jovin DJ, Subramaniam V, Jovin TM. 2002. Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM). Biophys J 83(3):1631–1649. 19. Clegg RM, Feddersen B, Gratton E, Jovin TM. 1992. Time-resolved imaging fluorescence microscopy. Proc SPIE 1640:448–460. 20. Wouters FS, Bastiaens PIH. 1999. Fluorescence lifetime imaging of receptor tyrosine kinase activity in cells. Curr Biol 9:1127–1130. 21. Verveer PJ, Squire A, Bastiaens PIH. 2001. Improved spatial discrimination of protein reaction states in cells by global analysis and deconvolution of fluorescence lifetime imaging microscopy data. J Microsc 202(3):451–456. 22. Bastiaens PIH, Squire A. 1999. Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell. Trends Cell Biol 9:48–52. 23. Pepperkok R, Squire A, Geley S, Bastiaens PIH. 1999. Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy. Curr Biol 9:269–272. 24. Dowling K, Hyde SCW, Dainty JC, French PMW, Hares JD. 1997. 2-D fluorescence lifetime imaging using a time-gated image intensifier. Opt Commun 135:27–31. 25. Oida T, Sako Y, Kusumi A. 1993. Fluorescence lifetime imaging microscopy (flimscopy). Biophys J 64:676–685. 26. Schneckenburger H, Konig K, Dienersberger T, Hahn R. 1994. Timegated microscopic imaging and spectroscopy in medical diagnosis and photobiology. Opt Eng 33(8):2600–2605. 27. Straub M, Hell SW. 1998. Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope. Appl Phys Lett 73(13):1769–1771. 28. Elangovan M, Day RN, Periasamy A. 2002. Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell. J Microsc 205(1):3–14. 29. Sharman KK, Periasamy A, Ashworth H, Demas JN, Snow NH. 1999. Error analysis of the rapid lifetime determination method for double-exponential decays and new windowing schemes. Anal Chem 71:947–952. 30. Vereb G, Jares-Erijman E, Selvin PR, Jovin TM. 1998. Temporally and spectrally resolved imaging microscopy of lanthanide chelates. Biophys J 74:2210–2222. 31. Herman B, Wodnicki P, Kwon S, Periasamy A, Gordon GW, Mahajan N, Wang XF. 1997. Recent developments in monitoring cal-
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 753 cium and protein interactions in cells using fluorescence lifetime microscopy. J Fluoresc 7(1):85–91. 32. Schneckenburger H, Gschwend MH, Strauss WSL, Sailer R, Steiner R. 1997. Time-gated microscopic energy transfer measurements for probing mitochondrial metabolism. J Fluoresc 7(1):3–10. 33. Webb SED, Gu Y, Lévêque-Fort S, Siegel J, Cole MJ, Dowling K, Jones R, French PMW, Neil MAA, Juskaitis R, Sucharov LOD, Wilson T, Lever MJ. 2002. A wide-field time-domain fluorescence lifetime imaging microscopy with optical sectioning. Rev Sci Instrum 73(4):1898–1907. 34. Dowling K, Dayel MJ, Hyde SCW, French PMW, Lever MJ, Hares JD, Dymoke-Bradshaw AKL. 1999. High resolution time-domain fluorescence lifetime imaging for biomedical applications. J Mod Opt 46(2):199–209. 35. Cole MJ, Siegel J, Webb SED, Jones R, Dowling K, Dayel MJ, Parsons-Karavassilis D, French PMW, Lever MJ, Sucharov LOD, Neil MAA, Juskaitis R, Wilson T. 2001. Time-domain whole-field fluorescence lifetime imaging with optical sectioning. J Microsc 203(3):246–257. 36. Kentech Instruments Ltd., Oxfordshire, UK. http://www.kentech. co.uk. 37. LaVision Inc., Ypsilanti, Michigan. http://www.lavision.de/products/ cameras/picostar/ picostar.htm. 38. Hamamatsu Corp. http://usa.hamamatsu.com. 39. Diaspro A. 2002. Confocal and two-photon microscopy foundations, applications, and advances. Wiley-Liss, New Jersey, 56. 40. Duncan RR, Bergmann A, Cousin MA, Apps DK, Shipston MJ. 2004. Multi-dimensional time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to detect FRET in cells. J Microsc 215(1):1–12. 41. Becker & Hickl GmbH, Berlin. http://www.becker-hickl.de. 42. Becker W, Hickl H, Zander C, Drexhage KH, Sauer M, Siebert S, Wolfrum J. 1999. Time-resolved detection and identification of single analyte molecules in microcapillaries by time-correlated singlephoton counting (TCSPC). Rev Sci Instrum 70(3):1835–1841. 43. Becker W, Bergmann A. 2004. Multi-dimensional time-correlated single photon counting. In Review in fluorescence 2005. Ed CD Geddes, JR Lakowicz. Springer, New York. 44. Becker W, Bergmann A, Hink MA, Konig K, Benndorf K, Biskup C. 2004. Fluorescence lifetime imaging by time-correlated single-photon counting. Microsc Res Technol 63:58–66. 45. Becker W, Bergmann A, Weiss G. 2002. Lifetime imaging with the Zeiss LSM-510. Proc. SPIE 4620:30–35. 46. Becker W, Bergmann A, Biscotti G, Koenig K, Riemann I, Kelbauskas L, Biskup C. 2004. High-speed FLIM data acquisition by time-correlated single photon counting. Proc SPIE 5323:27–35 47. Becker W, Bergmann A, Biscotti G, Ruck A. 2004. Advanced timecorrelated single photon counting technique for spectroscopy and imaging in biomedical systems. Proc SPIE 5340:104–112 48. van der Oord CJR, de Grauw CJ, Gerritsen HC. 2001. Fluorescence lifetime imaging module LIMO for CLSM. Proc SPIE 4252:119–123. 49. Becker W, Bergmann A. 2002. Multi-wavelength TCSPC lifetime imaging. Proc SPIE 4620:79–84. 50. Becker W, Bergmann A, Konig K, Tirlapur U. 2001. Picosecond fluorescence lifetime microscopy by TCSPC imaging. Proc SPIE 4262:414–419. 51. Köllner M, Wolfrum J. 1992. How many photons are necessary for fluorescence-lifetime measurements? Chem Phys Lett 200:199–204. 52. Becker & Hickl GmbH, Berlin, Germany, http://www.beckerhickl.de, 2000. TCSPC adds a new dimension to 3D laser scanning microscopy. Photonik 3:16–19 53. Berezovska O, Ramdya P, Skich J, Wolfe MS, Bacskai BJ, Hyman BT. 2003. Amyloid precursor protein associates with a nicastrindependent docking site on the presenilin 1-(-secretase complex in cells demonstrated by fluorescence lifetime imaging. J Neurosci 23(11):4560–4566. 54. Carlsson K, Liljeborg A. 1998. Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiplewavelength scanning (IMS) technique. J Microsc 191(pt.2):119–127. 55. Gratton E, Breusegem S, Sutin J, Ruan Q, Barry N. 2003. Fluorescence lifetime imaging for the two-photon microscope: timedomain and frequency-domain methods. J Biomed Opt 8(3):381– 390. 56. Carlsson K, Liljeborg A, Andersson RM, Brismar H. 2000. Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance. J Microsc 119(pt.2):106–114. 57. Hanson KM, Behne MJ, Barry NP, Mauro TM, Gratton E, Clegg RM. 2002. Two photon fluorescence lifetime imaging of the skin stratum corneum pH gradient. Biophys J 83:1682–1690. 58. Molecular Probes product information FluoCels #1. http://www. probes.com/ servlets/product?item-14780. ADDITIONAL READING ON FLUORESCENCE- LIFETIME IMAGING MICROSCOPY Applications of FLIM Calleja V, Ameer-Beg SM, Vojnovic B, Woscholski R, Downward J, Larijani B. 2003. Monitoring conformational changes of proteins in cells by fluorescence lifetime imaging microscopy. Biochem J 372:33–40. Eliceiri KW, Fan C-H, Lyons GE, White JG. 2003. Analysis of histology specimens using lifetime multiphoton microscopy. J Biomed Opt 8(3):376–380. Harpur AG, Wouters FS, Bastiaens PIH. 2001. Imaging FRET between spectrally similar GFP molecules in single cells. Biotechnology 19: 167–169. Marriott G, Clegg RM, Arndt-Jovin DJ, Jovin TM. 1991. Time resolved imaging microscopy, phosphorescence and delayed fluorescence imaging. Biophys J 60:1374–1387. Tadrous PJ, Siegel J, French PMW, Shousha S, Lalani E-N, Stamp GWH. 2003. Fluorescence lifetime imaging of unstained tissues: early results in human breast cancer. J Pathol 199:309–317. van Zandvoort MAMJ, de Grauw CJ, Gerritsen HC, Broers JLV, oude Egbrink MGA, Ramaekers FCS, Slaaf DW. 2002. Discrimination of DNA and RNA in cells by a vital fluorescent probe: lifetime imaging of SYTO13 in healthy and apoptotic cells. Cytometry 47:226–235. Vermeer JEM, Van Munster EB, Vischer NO, Gadella TWJ. 2004. Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy. J Microsc 214(2):190–200.
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Principles of Fluorescence Spectros
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Joseph R. Lakowicz Center for Fluor
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Preface The first edition of Princi
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x GLOSSARY OF ACRONYMS DNS dansyl o
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xii GLOSSARY OF ACRONYMS TRES time-
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xiv GLOSSARY OF MATHEMATICAL TERMS
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xvi CONTENTS 2.9.4. Conversion betw
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xviii CONTENTS 6. Solvent and Envir
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xx CONTENTS 10.4.4. Alignment of Po
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xxii CONTENTS 14.7.1. Simulations o
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xxiv CONTENTS 19.12. New Approaches
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xxvi CONTENTS 25.3. Optical Propert
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2 INTRODUCTION TO FLUORESCENCE Figu
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6 INTRODUCTION TO FLUORESCENCE inst
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10 INTRODUCTION TO FLUORESCENCE Fig
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12 INTRODUCTION TO FLUORESCENCE ns,
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14 INTRODUCTION TO FLUORESCENCE 1.7
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24 INTRODUCTION TO FLUORESCENCE due
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28 INSTRUMENTATION FOR FLUORESCENCE
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3 Fluorescence probes represent the
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98 TIME-DOMAIN LIFETIME MEASUREMENT
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100 TIME-DOMAIN LIFETIME MEASUREMEN
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238 DYNAMICS OF SOLVENT AND SPECTRA
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278 QUENCHING OF FLUORESCENCE 8.1.
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280 QUENCHING OF FLUORESCENCE Figur
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282 QUENCHING OF FLUORESCENCE ly ev
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290 QUENCHING OF FLUORESCENCE relat
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298 QUENCHING OF FLUORESCENCE σ ar
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320 QUENCHING OF FLUORESCENCE 47. R
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322 QUENCHING OF FLUORESCENCE 113.
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328 QUENCHING OF FLUORESCENCE P8.3.
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330 QUENCHING OF FLUORESCENCE P8.11
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332 MECHANISMS AND DYNAMICS OF FLUO
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10 Measurements of fluorescence ani
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444 ENERGY TRANSFER Figure 13.1. Fl
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446 ENERGY TRANSFER wavelength is e
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448 ENERGY TRANSFER Table 13.1. Cal
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450 ENERGY TRANSFER Figure 13.6. Ab
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452 ENERGY TRANSFER safe for many p
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462 ENERGY TRANSFER tiple acceptors
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468 ENERGY TRANSFER Table 13.3. Rep
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470 ENERGY TRANSFER 55. Clegg RM. 1
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472 ENERGY TRANSFER Tong AK, Jockus
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474 ENERGY TRANSFER Figure 13.39. O
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14 In the previous chapter we descr
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16 The biochemical applications of
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578 TIME-RESOLVED PROTEIN FLUORESCE
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19 Fluorescence sensing of chemical
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676 NOVEL FLUOROPHORES Figure 20.1.
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678 NOVEL FLUOROPHORES Figure 20.7.
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680 NOVEL FLUOROPHORES Figure 20.12
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698 NOVEL FLUOROPHORES 33. Acherman
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Appendix I Corrected Emission Spect
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Appendix II Fluorescence Lifetime S
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890 APPENDIX III P ADDITIONAL READI
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892 APPENDIX III P ADDITIONAL READI
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894 ANSWERS TO PROBLEMS A1.4. A. Th
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896 ANSWERS TO PROBLEMS Energy tran
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898 ANSWERS TO PROBLEMS Figure 4.65
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900 ANSWERS TO PROBLEMS expected to
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Index A Absorption spectroscopy, 12
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Principles of Fluorescence Spectroscopy

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