The division has a custom made setup for high-resolution fluorescence imaging capable of single molecule studies, fluorescence recovery after photobleaching (FRAP) and total internal reflection fluorescence (TIRF) imaging at two wavelengths simultaneously (488 nm and 638 nm excitation). The setup is also combined with a piezo-controlled (with nm resolution) holder for nano- and micropipette manipulation and delivery. The setup is based around a Nikon Eclipse TE2000 U microscope, with a Nikon Apo TIRF 60x (NA = 1.49) oil objective for the TIRF studies. Other objectives available are: 10x, 20x, and 60x air objectives. Excitation light comes from two Cobolt MLD diode lasers operating at 488 nm (maximum 60 mW power) and 638 nm (maximum 140 mW power). The excitation laser power is adjusted with a motorized filter wheel from Thorlabs. Images are collected with a sCMOS camera from Hamamatsu (ORCA-Flash4.0 LT). The software used to control the setup is the open source program Micro-Manager. Funding from the Royal Physiographic Society in Lund, the Royal Swedish Society, Per-Eric and Ulla Schyberg’s Foundation, and the Crafoord Foundation is gratefully acknowledged.