1 Summary ...................................................... 3
2 Fluctuation methods .......................................... 5
2.1 Introduction ............................................ 5
2.2 Fluorescence fluctuation methods, FCS ................... 6
2.2.1 Principle and experimental realization ........... 6
2.2.2 Theory ........................................... 8
2.2.3 Extensions of FCS and combination with other
techniques ...................................... 11
2.2.4 Practical aspects ............................... 11
3 Spatio-temporal correlations ................................ 13
3.1 Fixed configuration .................................... 13
3.2 Scanning FCS ........................................... 14
4 Two-photon excitation ....................................... 19
4.1 Principle .............................................. 19
4.2 Two-photon excitation in fluorescence microscopy ....... 19
4.2.1 Two-photon excitation in FCS .................... 20
4.2.2 Saturation ...................................... 20
4.3 Photobleaching ......................................... 22
4.3.1 Photobleaching mechanisms ....................... 24
4.3.2 Reduction of photobleaching ..................... 25
5 Biophysical applications of fluctuation methods ............. 27
6 Perspectives ................................................ 29
7 References .................................................. 32
8 Included articles ........................................... 41
A1 Fluctuations as a source of information in fluorescence
microscopy .................................................. 41
A2 Scanning fluorescence correlation spectroscopy .............. 55
A3 Simultaneous two-photon fluorescence correlation
spectroscopy and lifetime imaging of dye molecules in
submicrometer fluidic structures ............................ 81
A4 Precise measurement of diffusion coefficients using
scanning fluorescence correlation spectroscopy .............. 91
A5 Circular scanning fluorescence correlation spectroscopy on
membranes ................................................. 105
|
