Supplementary Fig. 1: Iso-LFM setup.
From: Instantaneous isotropic volumetric imaging of fast biological processes
(a) Schematic 2D drawing of the setup showing the main opto-mechanical components. Note that the two imaging objectives do not face each other co-axially but are arranged in an orthogonal fashion as further indicated in (c). Depending on the excitation beam size (adjustable beam expander) and scan mode of the galvanometric mirrors either a conventional light-sheet (yellow plane), or a selective volume can be illuminated (b). For very fast imaging modes (50 Hz or more), we chose to overfill a quadratic 2D slit which was relayed into the focal volume of the detection objectives, in order to instantaneously excite the entire volume of interest. The fluorescence is detected by two orthogonally oriented detection paths. A narrow bandpass-filter (BP filter - Semrock, 525/50 nm BrightLine) is placed in front of the tube lens, which focuses either on the camera sensor (light sheet mode) or on the microlens array (Iso-LFM mode). In the latter case, the camera sensor is displaced by one focal length of the microlenses further away from the tube lens. A 1-1 relay lens system is used to relay the image plane of the camera sensor (see Methods for details). (c) Experimental implementation of the microscope. Optical paths are colored as in (a). Zoom-in shows a close-up of the three mutually orthogonal objectives and the sample area.
