Confocal microscope significantly expands the capabilities of multi-user facility and complements a number of other Leica systems already used on site
In order to provide increased versatility and sensitivity when undertaking fluorophore-based live cell imaging applications, the cell imaging facility at Bristol University has installed a Leica Microsystems TCS SP2 AOBS microscope.
This filter-free spectral confocal microscope will enable researchers to efficiently perform multiple GFP experiments including Fret, Frap and Flip protocols.
The new microscope significantly expands the capabilities of this multi-user facility and complements a number of other Leica systems already used on site.
Since the cell imaging facility provides core support and a focal point for interactions between many different research groups, the versatility of the SP2 AOBS played a key role in the university's selection process.
In addition to simultaneously imaging up to four fluorescent proteins through its enhanced transmittance, the SP2 AOBS also has input ports for additional lasers, external detectors for non-descanned imaging and output ports for Flim detectors or spectrometers.
The SP2 AOBS has proved particularly useful to Bristol University through its ability to separate even closely overlapping fluorophores, such as CFP, GFP and YFP.
"The multi-channel imaging made possible by the SP2 AOBS's unique beam splitter set up has made it much easier for us to undertake simultaneous imaging of a wide variety of fluorescent proteins and dyes," said Mark Jepson, research fellow and manager of the cell imaging facility. "The availability of the 405nm laser has also proved very useful for Dapi imaging and photoactivatable GFP applications".
The sensitivity of the SP2 was also a key consideration, since it is approximately 30% more sensitive than conventional confocal microscopes that use glass-based dichroic mirrors.
This is again due to its proprietary electronically controlled and freely definable beam splitter at the heart of its multiple fluorophore imaging capabilities.
The Leica TCS SP2 AOBS is the first confocal system to allow high precision recording of fluorescence spectra without modulation by splitters or filters. "We are particularly impressed by the sensitivity of the SP2 AOBS which crucially allows us to reduce the amount of excitation light when working with living specimens," said Jepson. "In addition, the system's ability to utilise its quality hardware, rather than relying on software, to enhance fluorescence separations enables us to optimise separation without compromising on resolution or acquisition rate." The cell imaging facility is also using the visible and UV confocal capabilities of the SP2 AOBS for a range of other applications that deliver still, movie and 3D images.
