By Melbourne Centre for Nanofabrication
A Laser Total Internal Reflection Fluorescence (TIRF) microscope allows users to observe very thin sections of a material, usually around 200nm thick, which are held in an aqueous solution on the surface of a glass slide. A TIRF microscope aims to selectively illuminate and excite cells in contact with the surface of the glass slide through the use of a laser. As the specimens viewed on a TIRF microscope are so thin, there is minimal noise behind the cells which have been illuminated, making it possible to view single molecule fluorescence. TIRF Microscopy is used to perform investigations in living cells and to image minute structures or single molecules. It can be used to perform nanoparticle array characterisation and to view cell functions such as exocytosis and protein trafficking. The Nikon Instrument TIRF with Ti-U system is attached to an inverted Nikon Microscope employing class 3B lasers for angle measurements. It features three solid state lasers of 405nm, 488nm and 561nm to cover fluorescent dyes of blue to red.
