Optics at the Nanometer Scale pp 223-233 | Cite as
Fluorescence Lifetime Contrast Combined with Probe Microscopy
Chapter
Abstract
Fluorescence lifetime imaging is combined with atomic force microscopy in an integrated scanning microscope using a silicon-nitride probe. The time decay of fluorescence is measured at each image position with a resolution of 50 ps by time correlated single photon counting using a frequency doubled mode-locked Ti-Sapphire laser and fast electronics. Images of a mixture of fluorescence labelled latex spheres are presented where the lifetime contrast of the different spheres can be directly correlated to the topography as detected by force microscopy.
Keywords
Fluorescence Lifetime Fluorescence Decay Fluorescence Lifetime Image Microscope Single Exponential Decay Latex Sphere
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Preview
Unable to display preview. Download preview PDF.
References
- 1.Gadella, T.W., Jovin, T.M. and Clegg, R.M. (1993) Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the ns time scale, Bioph. Chem. 48, 221–239.CrossRefGoogle Scholar
- 2.Lakowicz, J.R. and Berndt, K.W. (1991) Lifetime-selective fluorescence imaging using an RF phase sensitive camera, Rev. Sci. Instrum. 62, 1727–1734.ADSCrossRefGoogle Scholar
- 3.Laczko, G., Gryczynski, I., Gryczynski, Z., Wiczk, W., Malak, H. and Lakowicz, J.R. (1990) Phase fluorometry. Rev. Sci. Instrum. 61, 2331.ADSCrossRefGoogle Scholar
- 4.Buurman, E.P., Sanders, R., Draaijer, A., Gerritsen, H.C., van Veen, J.J.F., Houpt, P.M. and Levine, Y.K. (1992) Fluorescence lifetime imaging using a confocal laser scanning microscope, Scanning 14, 155–159.CrossRefGoogle Scholar
- 5.Betzig, E. and Trautman, J.K. (1992) Near-field optics: microscopy, spectroscopy and surface modification beyond the diffraction limit, Science 257, 189–195.ADSCrossRefGoogle Scholar
- 6.Betzig, E. and Chichester, R.J. (1993) Single Molecule observed by Near field Scanning Optical Microscopy, Science 262, 1422–1425.ADSCrossRefGoogle Scholar
- 7.Trautman, J.K., Mackiin, J.J., Brus, L.E. and Betzig, E. (1994) Near field spectroscopy of single molecules at room temperature, Nature 369, 40–42.ADSCrossRefGoogle Scholar
- 8.Dunn, R.C., Holtom, G.R., Mets, L. and Sunney Xie, X. (1994), Near-field fluorescence imaging and fluorescence lifetime measurement of light harvesting complexes in intact photosynthetic membranes, J. Phys. Chem. 98, 3094–3098.CrossRefGoogle Scholar
- 9.Sunney Xie, X. and Dunn, R.C. (1994) Probing Single Molecule Dynamics, Science 265, 361–364.ADSCrossRefGoogle Scholar
- 10.Ambrose, W.P., Goodwin, P.M., Martin, J.C. and Keller, R.A. (1994) Alterations of Single Molecule Fluorescence Lifetimes in Near-Field Optical Microscopy, Science 265, 364–367.ADSCrossRefGoogle Scholar
- 11.Van Hülst, N.F., Moers, M.H.P., Noordman, O.F.J., Faulkner, T., Segerink, F.B., van der Werf, K.O., De Grooth B.G. and Böiger B. (1992) Operation of a Scanning Near field Optical Microscope in reflection in combination with a scanning force microscope, SPIE 1639, 36–43.Google Scholar
- 12.Moers, M.H.P., Tack, R.G., Van Hülst, N.F. and Böiger, B. (1994) Photon scanning tunneling microscope in combination with a force microscope, J. Appl. Phys. 75, 1254–1257.ADSCrossRefGoogle Scholar
- 13.O’Connor, D.V. and Philips, D. (1984) Time correlated single photon counting, Academic Press, LondonGoogle Scholar
- 14.Wilkerson, C.W. Goodwin, P.M., Ambrose, W.P., Martin, J.C. and Keller, R.A. (1993) Detection and lifetime measurement of single molecules in flowing sample streams by laser-induced fluorescence, Appl. Phys. Lett. 62, 2030–2032.ADSCrossRefGoogle Scholar
- 15.Van Hülst, N.F., Moers, M.H.P. and Böiger, B. (1993) Near-field optical microscopy in transmission and reflection modes in combination with force microscopy, J. Microscopy. 171, 95–105.CrossRefGoogle Scholar
- 16.Ruiter, A.G.T., Moers, M.H.P., Jalocha, A. and van Hülst, N.F. (1995) Development of an integrated NSOM probe. Ultramicroscopy 58, in press.Google Scholar
- 17.Grabowski, J. and Gantt, E. (1978) Photochemistry and Photobiology 28, 39.CrossRefGoogle Scholar
- 18.Chen, R.F. (1969) Archives of Biochemistry and Biophysics 133, 263CrossRefGoogle Scholar
- 19.Haughland, R.P. (1992) Handbook of fluorescent probes and research chemicals, 5th ed., Molecular Probes Inc. Eugenen, Oregon.Google Scholar
Copyright information
© Kluwer Academic Publishers 1996