Located in room D28 of the Lyman Duff Medical Building, the Department of Microbiology and Immunology is equipped with a Fluoview 1000 laser-scanning confocal microscope and associated computing platform from Olympus. The FV1000 microscope in our department contains 458/488/515/543/633 nm wavelength lasers allowing for simultaneous multi-colour imaging, 10-100x objectives, live imaging capability and has been recently upgraded (2014-2015) with high resolution gadolinium detectors, a mechanical stage and a new computer equipped with stitching software for whole organ image capture. In addition, the microscope has been recently been equipped with an LED light source for epifluorescent microscopy to meet a range of experimental needs.
Laser-scanning confocal microscopy is a technology that allows for high resolution, optical sectioning of individual cells in culture or embedded within tissues. The wide range of fluorescent probes and detectors enables rapid analysis of multiple parameters within one sample. The advantage of this technology over wide-field epifluorescence microscopy is that it uses point illumination to enhance image resolution and control the depth of the focal plane within tissues. The use of lasers of different wavelengths to scan tissues and excite fluorescent cell labels both horizontally and vertically allows 3D rendering of cells within whole tissues. Using this instrument, investigators can accurately measure cell size and study cell morphology, membrane and intracellular protein expression as well as cell division and death. A particular benefit of this technology is that it combines these cellular parameters with detailed information about the location of specific cell types within complex tissues. This information elucidates how intercellular interactions impact organ-specific states of health and disease. Check the tutorial on the basics of confocal immunofluorescence microscopy.
Appointments to use this instrument are taken using an on-line booking system. Services provided by this facility including training of new users to ensure sufficient background knowledge to successfully design experiments, operate the equipment properly and with confidence. The facility also offers technical consultation on experiment design and analysis.