Martin Oheim and Florian Schapper 2005 J. Phys. D: Appl. Phys. 38 R185 doi:10.1088/0022-3727/38/10/R01
Martin Oheim1,3 and Florian Schapper2
Show affiliationsTotal internal reflection fluorescence (TIRF), a general term that embraces any spectroscopic or microscopic technique based on the evanescent field created by TIR of light, is further establishing itself as an important tool for studying near-surface phenomena. Impingement of a femtosecond-pulsed infrared beam on a reflecting interface creates the conditions for 'macroscopic' evanescent-field two-photon fluorescence excitation. The two-photon fluorescence excitation volume is confined by both the non-linearity of the multi-photon process and the spatial inhomogeneity of the evanescent field. The absence of scattered excitation resulting in a low background and the possibility of simultaneous multi-colour fluorescence excitation should make non-linear evanescent-field excitation particularly attractive for quantitative single-molecule observation and ultra-sensitive screening assays. In this topical review, we survey the requirements, present the current results and explore the potential of this novel non-linear microscopy.
42.65.Re Ultrafast processes; optical pulse generation and pulse compression
07.60.Pb Conventional optical microscopes
42.25.Gy Edge and boundary effects; reflection and refraction
Issue 10 (21 May 2005)
Received 5 January 2005, in final form 15 March 2005
Published 6 May 2005
Martin Oheim and Florian Schapper 2005 J. Phys. D: Appl. Phys. 38 R185
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