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Fixed-stage upright multiphoton microscope equipped with a Spectra-Physics MaiTai eHP DeepSee laser (70 fs pulse width, 80 MHz repetition rate, continuously tunable between 690--1040 nm). Laser input power is software-controlled by means of an acousto-optic modulator, which also acts as a high-speed shutter. The objective is motorised with a z-axis resolution of 10 nm. Pixel dwell time can be set as low as 2 us (unidirectional scanning) and 0.5 us (bidirectional). Scanning modes include 2D raster, 3D stack, straight line, freehand line and single point. 2D and 3D scans can also be performed as a function of time. Motorised xy-stage is joystick- and software controlled, and allows for multi-area time-lapse experiments.The microscope is equipped with four non-descanned PMTs, two sub- and two superstage. One of the superstage detectors has been customised for detecting fluorescence at wavelengths longer than the usual 690 nm. There is also an additional substage PMT for the confocal detection of transmitted laser light (Dodt gradient contrast).Wide-field imaging modes include transmitted white light (Dodt contrast) and epifluorescence, which can be observed using laser-safe oculars or a 3.3 MP colour CMOS camera . Electrophysiology experiments can be carried out using two triple-axis micromanipulators (Scientifica) and a MultiClamp 700B amplifier connected to a Digidata 1550 digitiser (both from Molecular Devices). |
