Studying Cell–Surface Interactions In Vitro: A Survey of Experimental Approaches and Techniques - Tissue Engineering: Cell-Surface Interactions for Tissue Culture - page 44

Fig. 5 a Schematic illustration of image formation in TIRF/TIRAF microscopy. b Typical
TIRAF image of the cell-substrate junction (adapted from [ 27 ])
Fig. 6 a Kretschmann configuration for performing integral SPR analysis of the cell-surface
junction. Surface plasmon excitation is recorded as a dip of the reflectivity as a function of
incident angle. During kinetic measurements the changes in reflectivity are acquired at a constant
angle of incidence. b Setup for SPR imaging based on a high numerical aperture objective
approaches for sensitive and time-resolved analyses of cell-substrate interactions.
Most of these approaches are based on the so-called Kretschmann configuration in
which the excitation light is coupled into the metal surface via a high refractive
index prism for spectroscopic (Fig. 6 a) or a high numeric aperture objective for
microscopic applications (Fig. 6 b). Spectroscopic analyses are performed by either
measuring the changes in reflectivity at a constant (monochromatic) excitation
wavelength but variable angle of incidence (angle-dependent mode) or by
measuring the reflectivity at a constant angle of incidence but with polychromatic
excitation and subsequent spectral analysis of the reflected light (wavelength-
dependent mode) [ 29 ]. In order to perform SPR-based microscopy, a collimated
monochromatic light path is used to excite the whole field of view and the reflected