Total Internal Reflection Fluorescence (TIRF) allows the excitation of molecules within a range of around 150nm from the coverslip. The resulting images are very contrast-rich and background-free. TIRF is very useful to study processes like adhesion, membrane homeostasis, or endo- and exo-cytosis.
Olympus TIRF Setup
The Olympus TIRF setup is the only commercially available system, which allows individual manipulation of the TIRF angle for four laser lines. This provides the possibility to observe several fluorescences simultaneously at custom chosen TIRF penetration depths. The setup is equipped with an inverted, mostly manual microscope, hardware based focus drift-correction, and two high-sensitive EM-CCD cameras to acquire simultaneous images. The setup provides four excitation lasers (405 nm, 473 nm, 561 nm, 633 nm). In addition, the system has a stabilized advanced widefield fluorescence illumination (MT20). A small incubation chamber for temperature and CO2 control is available as well