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Bioimaging Facility: Advanced Light Microscopy

The state of the art Bioimaging and Microscopy research and teaching facility in the Department of Biosciences is at the core of the Durham Centre for Bioimaging Technology and houses an extensive selection of imaging modalities

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Leica SP5 LSCM SMD: Tandem Scanner, FLIM & FCCS

Location: Biosciences: Room 137

This confocal is also capable of Fluorescence Lifetime Imaging Microscopy and Fluorescence Cross Correlation Spectroscopy.

The Leica SP5 is a fast broadband confocal, capable of a full range of scan speeds at the highest resolution. The high-efficiency SP detection system (five channels simultaneously) and the fast AOBS (Acousto Optical Beam Splitter, instead of dichroic mirrors), means this system can produce bright, noise-free images with minimal photo damage at high speed. In the SP detection system, instead of filters, spectral separation is performed by a prism and adjustable mirror-sliders in front of each detector. Importantly this arrangement allows simultaneous data acquisition at five individually tuneable spectral bands. This tunability is particularly useful when working with novel fluorophores and when discrimination of autofluorescence is required. In addition to PMTs the microscope also has a HyD detector. This is a GaAsP photocathode which offers super sensitivity combined with a large dynamic range. Such detectors reduce acquisition times and allow the use of lower laser intensities giving reduced photobleaching, phototoxicity and improved cell viability. The combination of HyD detection with PMTs in our system offers a dynamic range from single photon counting to whole organism imaging. This system is not only capable of high quality images (frame sizes up to 8192 x 8192 pixels, 64 Mpix), but can also acquire at speed. The tandem scanner contains a conventional and resonant scanner which can easily be switched between. The resonant scanner enables you to acquire high speed images at large formats (full field high speed imaging) with line-frequencies up to 16 kHz (512x512 image at 25fps) very fast for a point scanner.

Our SP5 has been upgraded with the Leica SMD (Single Molecule Detection) package introducing FLIM and FCCS capabilities.

The microscope in enclosed in an environmental chamber allowing the control of CO2 and temperature.

This system is good for: Live cell imaging, high quality, high speed.

Bioimaging Home
SP5 crop for spec

Lens

Numerical Aperture
Leica 10x HCX PL APO CS 0.4
Leica 20x HCX PL APO Oil UV 0.7
Leica 40x HCX PL APO Oil UV 1.25 
Leica 63x HCX PL APO Oil UV 1.4
Leica 63x HCX PL APO CS Water 1.2

 

Laser LinesEmission Filters
405nm Fully tunable, up to 5 simultaneously
458nm, 478nm, 488nm, 494nm, 514nm  
543nm  
594nm  
633nm  

 

FLIM Laser LinesEmission Filters
470nm pulsed Spectral FLIM PMTs fully tunable or
640nm pulsed 2x external SPADs GFP/RFP or FITC/TRITC
Bioimaging Home
SP5 crop for spec