My post-doctoral work involved researching protein targeting and flagellum assembly in the protozoan parasite, Trypanosoma brucei. T. brucei are microscopic protozoan parasites that cause African Trypansomasis, known as sleeping sickness in humans and Nagana in cattle. Disease is spread by infected tsetse flies and is endemic in parts of sub-Saharan Africa. Our interest lies in the cell biology of these parasites not only because of their medical and veterinary importance but as they are also an experimentally tractable organism with which to study the formation of the eukaryotic cilium/flagellum. Cilia are evolutionarily conserved structures and as many proteins required for its assembly are also in Trypanosomes they can be used as models with which to study human ciliopathic disease.

The BLS Bioimaging facility houses two fluorescence microscope, a Zeiss LSM 510 Meta laser scanning confocal and an Applied Precision DeltaVision deconvolution microscope.

Confocal microscopy is particularly suited for applications that require high axial resolution (thin optical sections) and offers the advantage of increasing image resolution by removing out of focus information. Our Zeiss LSM 510 Meta is an upright confocal, with a fully motorised stage, 2PMTs and a Meta head for the acquisition of emission spectra. It has seven laser lines to choose from; UV 405nm at 25mW, Argon 458/477/488/514nm at 30mW, HeNe 543nm at 1mW and HeNE 633 at 3mW.

Widefield deconvolution microscopy combines a high quality epifluorescence microscope with the computational ability to increase the contrast and resolution of a digital image through software processing to ‘re-focus’ out of focus pixels of light. When compared to confocal imaging, image acquisition time on a widefield microscope is much reduced. Our Applied Precision inverted DeltaVision has a fully motorised stage, 100W Hg arc lamp, filter blocks for excitation and detection of DAPI, FITC, TRITC/Rhodamine, CFP and YFP and a high resolution Cool Snap-HQ camera. We also have an environmental chamber with stringent heat and CO₂ controls to increase cell viability for live-cell imaging. 

BIOL272 Cell Biology Techniques: Microscopy