Fast FRET! Image capture at two wavelengths simultaneously

20 Oct 2008
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Faster, simpler imaging of cellular events is now possible as Nikon’s Eclipse Ti Series inverted microscopes can capture images at two different wavelengths simultaneously, using dual cameras. Accelerating image acquisition while maintaining full frame resolution, the system is ideal for FRET and the capture of rapid dynamic cellular events using calcium or other ion-targeted probes, ratio probes, dual emission ratiometric dyes etc.

The two cameras are positioned on the Eclipse Ti’s back and side ports. Perfect registration between the two cameras is assured on installation to ensure that no information is lost during imaging. No further realignment or specialised alignment software is required. Even when the intensity difference between wavelengths is large, high-quality images can be captured by adjusting camera sensitivity for each wavelength.

Nikon has partnered with Andor to offer their full range of high-performance iXon+ and Luca EMCCD cameras. The IxonEM+ 897 back-illuminated EMCCD camera offers high sensitivity, low noise and rapid frame rates giving distinct speed advantages in FRET applications. The cameras are optimised for use with Nikon’s dedicated NIS-Elements software for image capture, processing and analysis. Unified integrated control of microscope and cameras offers significant benefits for cutting-edge live cell research. NIS-Elements C for confocal microscope applications includes FRET analysis software as standard.

The dual camera system has obvious advantages when compared with single camera acquisition, allowing simultaneous full frame capture of two different wavelengths without dividing the CCD chip and compromising image resolution; and removing the need for filter switching, which normally constrains speed of sequential image acquisition or can lose valuable information.

The Nikon Eclipse Ti series offers improved system speed, increased flexibility and efficient multi-mode microscopy; and, coupled with Andor specialised iXon+ cameras, provides a fully-integrated microscope system ideal for high-end research and rapid live cell imaging.

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