Due to their unique properties-coherent radiation, diffraction limited focusing, low spectral bandwidth and in many cases short light pulses-lasers play an increasing role in live cell microscopy. Lasers are indispensable tools in 3D microscopy, e.g., confocal, light sheet or total internal reflection microscopy, as well as in super-resolution microscopy using wide-field or confocal methods. Further techniques, e.g., spectral imaging or fluorescence lifetime imaging (FLIM) often depend on the well-defined spectral or temporal properties of lasers. Furthermore, laser microbeams are used increasingly for optical tweezers or micromanipulation of cells. Three exemplary laser applications in live cell biology are outlined. They include fluorescence diagnosis, in particular in combination with Förster Resonance Energy Transfer (FRET), photodynamic therapy as well as laser-assisted optoporation, and demonstrate the potential of lasers in cell biology and-more generally-in biomedicine.
Keywords: FLIM; FRET; TIRFM; confocal microscopy; laser micromanipulation; light sheet microscopy; optical tweezers; photodynamic therapy; spectral imaging; super-resolution.