We show that with an appropriately configured scanning quartz pipette coated with aluminum, a near-field scanning optical microscope (NSOM) can be constructed to operate in aqueous solution for applications in biology. Many of the technical limitations associated with a scanning pipette were circumvented by introducing a small modulation of the distance between the pipette and the sample. We show that this ac method allows the pipette to be positioned very close to the sample surface and is robust in obtaining reproducible NSOM images in solution. This approach is also compatible with fluorescence imaging and fluorescence resonance energy transfer, and should further facilitate the use of NSOM in various areas of cell biology where high resolution is considered to be critical.
Validerad; 2001; 20070410 (ysko)