Fluorescence correlation spectroscop

With the aim of elucidating molecular dynamics in a small domain, we have constmcted several microspectroscopic systems, that is, (i) the confocal microscope with the excitation light source being a femtosecond NIR laser emitting a 35 fs pulse, and (ii) the fluorescence correlation spectroscopic system with optical tweezers. 

Ito, S., Toitani, N., Pan, L., Tamai, N. and Miyasaka, H. (2007) Fluorescence correlation spectroscopic study on water-soluble cadmium telluride nanocrystals fast blinking dynamics in the ps-ms region. J. Phys. Condens. Matter, 19, 486208. 

Yet the relationship between solute chemical structure and diffusion is not always simple. Werner et al. [248] conducted fluorescence correlation spectroscopic studies of three fluorescent probes in l-butyl-3-methylimidazolium hexafluorophosphate. The probes were chosen to be of comparable molecular structure, but possessed positive, negative, and neutral charges. The authors found that while the neutral probe diffused more rapidly than the cationic probe, the anionic probe diffused the most quickly. 

However, in an attempt to integrate the SFA and spectroscopic techniques, the use of silver for optical interferometry has been seen as a drawback due to the fact that it precluded sufficient excitation source intensity to illuminate the buried interface. In order to circumvent this problem Mukhopadhyay and co-workers in an experimental set-up where the SFA was combined with fluorescence correlation spectroscopy (FCS) used, instead of silver, multilayer dielectric coatings that allowed simultaneous interferometry and fluorescence measurements in different regions of the optical spectrum [75]. Using this set-up they succeeded in measuring diffusion in molecularly thin films with singlemolecule sensitivity. 

Mefliods of study and data interpretation still require further work and refinement. Several experimental techniques are used, including microscopy (TEM, SEM) dynamic light scattering " using laser sources, goniometers, and digital correlators spectroscopic methods (UV, CD, fluorescence) fractionation solubility and viscosity measurements and acid-base interaction. "... 

The CPF approach gives quantitative reement with the experimental spectroscopic constants (24-25) for the ground state of Cu2 when large one-particle basis sets are used, provided that relativistic effects are included and the 3d electrons are correlated. In addition, CPF calculations have given (26) a potential surface for Cus that confirms the Jahn-Teller stabilization energy and pseudorotational barrier deduced (27-28) from the Cus fluorescence spectra (29). The CPF method has been used (9) to study clusters of up to six aluminum atoms. 

Such ambiguity and also the low structural resolution of the method require that the spectroscopic properties of protein fluorophores and their reactions in electronic excited states be thoroughly studied and characterized in simple model systems. Furthermore, the reliability of the results should increase with the inclusion of this additional information into the analysis and with the comparison of the complementary data. Recently, there has been a tendency not only to study certain fluorescence parameters and to establish their correlation with protein dynamics but also to analyze them jointly, to treat the spectroscopic data multiparametrically, and to construct self-consistent models of the dynamic process which take into account these data as a whole. Fluorescence spectroscopy gives a researcher ample opportunities to combine different parameters determined experimentally and to study their interrelationships (Figure 2.1). This opportunity should be exploited to the fullest. 

A spectroscopic technique that probes membrane fluidity can either directly measure mobility and order parameters for membrane constituents (NMR) or use probes (ESR, fluorescence). Some fluorescent and ESR probes are shown in Fig. 4. The connection between the rotational correlation time of a membrane embedded probe and the membrane fluidity can be illustrated using the example of a simple isotropic liquid, in which fluidity is merely a reciprocal viscosity ri and the rotational correlation time Xc for a molecule with a hydrodynamic volume V is given by the well-known Debye-Stokes-Einstein relation Xc = r VlkT, where k is the Boltzmann constant and T is the... 

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