Microscopes

Tender description

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 recently refurbished rooms. Access will be granted to members of Compare, as well as the Intravital Microscopy (IVM) user group. The specifications below 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). 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, super resolution microscopy methods such as 2D/3D PALM/dSTORM and single-molecule FRET. The system should be equipped with a monochromator for maximal flexibility and speed in the selection of the excitation wavelength, an image 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). 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 512 x 512 mode and >= 560 frames/s from a 128 x 128 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 chamber allowing both ultrarapid exchange of solutions (< 2 ms) and long superfusion times (up to 1 hour). 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 perfusion systems/imaging chambers/stage insets should be ideally shared and interchangeable between the two microscopes (see above). The software should also allow the real-time monitoring of ratiometric FRET traces and the control/recording of the triggers to/from the perfusion system(s).