Tender for the Supply and Installation of FRET/Ca2+ Imaging and 4-Camera TIRF Microscopes to the University of Birmingham

Descriptions

The FRET/Ca2+ imaging and 4-camera TIRF microscopes are to be purchased by the Institute of Metabolism and Systems Research (IMSR), UoB. Match funding will be provided through COMPARE (http://www.birmingham-nottingham.ac.uk/compare/index.aspx). The systems will be housed in the recently refurbished rooms of the Centre of Membrane Proteins and Receptors (COMPARE). Access will be granted to members of COMPARE, as well as the intravital microscopy (IVM) user group. The specification below specifications outline the requirements for: a. A FRET/Ca2+ imaging microscope, which must be capable of performing fast high-sensitivity ratiometric imaging and photometric measurements in cells expressing FRET reporters or loaded with Ca2+ indictors (e.g. Fura-2). The system should be equipped with a monochromator for maximal flexibility and speed in the selection of the excitation wavelength, a dual-camera beam splitter, a very high sensitivity camera (EMCCD or equivalent). In addition, it should be equipped with a photomultiplier-based ratiometric acquisition system allowing ultrafast kinetics measurements (up to 100 microsecond temporal resolution). The system should also be equipped with a caged incubator for temperature control and a laminar-flow superfusion system (or more if needed) with compatible imaging chamber allowing both ultrarapid exchange of solutions (< 2 ms) and long superfusion times (up to 1 hour). The perfusion systems/imaging chambers/stage insets should be ideally interchangeable and could be shared between the two microscopes, allowing maximal flexibility. The microscope should be controlled by a user-friendly software, allowing the real-time monitoring of ratiometric FRET and Ca2+ traces and the control/recording of the triggers to/from the perfusion system(s). b. A 4-channel 2D/3D TIRF microscope for the analysis of both fast and slow dynamic biological processes (from few milliseconds to 1 hour) in living cells by single-molecule microscopy/single-particle tracking, superresolution microscopy methods such as 2D/3D PALM/dSTORM and single-molecule FRET. The system should also allow the fast and simultaneous acquisition of FRET between CFP and YFP by ratiometric imaging and single-molecule acquisition in the red or far-red channel. The system should also ideally allow to perform FRAP and/or uncaging experiments. The system should be equipped with suitable solid-state lasers of adequate power, a TIRF illuminator providing homogenous illumination, a quadruple camera beam-splitter and four fast EMCCD cameras. The microscope should be controlled by a user-friendly software, allowing fast synchronised real-time acquisition from the four cameras (>= 56 frames/s in normal full-frame 512x512 mode and >= 560 frames/s from a 128x128 sub-array in crop mode). The system should also be equipped with a caged incubator for temperature control and a laminar-flow superfusion system (or more if needed) with compatible imaging chambe...