Nano Science and Technology Institute
Nanotech 2007 Vol. 2
Nanotech 2007 Vol. 2
Technical Proceedings of the 2007 NSTI Nanotechnology Conference and Trade Show, Volume 2
Chapter 6: Biosensors

Parallel two channel near and far field fluorescence microscopy

Authors:D. Verdes, M. Rabe and S. Seeger
Affilation:University of Zurich, CH
Pages:545 - 548
Keywords:spectroscopy and imaging, bio sensors
Abstract:We report a new two channel fluorescence microscopy technique for surface-generated fluorescence. The realized fluorescence microscope allows high resolution imaging of aqueous samples. The core element of the instrument is a parabolic mirror objective that is used to collect the fluorescence at large surface angles above the critical angle of the water/glass interface. An aspheric lens, incorporated into the solid parabolic element, is used for diffraction limited laser focusing and for collecting fluorescence at low angles with respect to the optical axis. By separated collection of the fluorescence emitted into supercritical and subcritical angles, two detection volumes strongly differing in their axial resolution are generated at the water/glass interface. The collection of supercritical angle fluorescence (SAF) (by the parabolic mirror) results in a strict surface confinement of the detection volume whereas collecting below the critical angle (by the aspheric lens) allows gathering the fluorescence emitted several microns deep inside the aqueous sample. Consequently, the signals from surface-bound and unbound diffusing fluorescent molecules can be obtained simultaneously.
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