Laser-sharp images obtained from multiple fluorescence probes within a sample, particularly in live cell biology, are captured faster with a single scan over the full spectral range
Nikon says that with its Digital Eclipse C1si spectral laser confocal system, the quality of images obtained from multiple fluorescence probes within a sample, particularly in live cell biology, are now dramatically improved.
This is because Nikon's new confocal technology acquires high resolution data, over a spectral range of 400-750nm, in a single scan.
This provides the researcher with super fast, high quality, and continuous spectral imaging content.
Traditionally, fluorescence solutions have difficulty in cleanly separating signals from fluorescing probes and sample auto-fluorescence, due to overlap of their fluorescence emission spectra.
Taking a step up to a confocal solution usually resolves this, but this is limited by collecting data at specific wavelengths only.
This can be especially troublesome in Fret microscopy where precise localisation of the source of the signal is required.
In addition to this, the time taken to capture such information can potentially hinder some research into dynamic cell activity.
The C1si system can uniquely and simultaneously acquire image data over a 320nm wide spectral range, in 10nm wavelength resolutions or over smaller ranges at 5nm or 2.5nm resolutions, in less than 1s at 512x256pxl.
This is possible by utilising a 32 PMT channel detector and not placing dependence on the confocal pinhole size.
Furthermore, the Digital Eclipse C1si employs a mechanism to prevent illuminating wavelengths from blinding the detector.
The emission path can be easily switched between brightfield, differential interference contrast, standard three-channel fluorescence detection and the high resolution spectral detector, allowing a single instrument to be used for the widest possible range of applications.
Uncompromised optical design and signal processing of the Digital Eclipse C1si system places it among the most efficient spectral detectors available, says Nikon.
The combination of the newly developed diffraction efficiency enhancement system (Dees), high efficiency fluorescence transmission technology, and superior Nikon VC objectives, optimise the signal reaching the photomultiplier tubes, thus making the image brighter and sharper.
The newly developed dual integration signal processing (Disp) technology eliminates digitisation dead time.
The combination of Disp and Dees technology ensures the highest efficiency spectral imaging with the best signal to noise ratio, says tghe company.
The power of the C1si is fully realised in modern research where there is a need to use a plethora of fluorochrome markers.
It can capture data over a wide spectral range encompassing fluorescence proteins such as CFP, GFP, YFP, DsRed in one hit, and conversely can cleanly separate the signals of close spectral emitters GFP and FITC.
The high sensitivity, fast image capture, and excellent signal to noise characteristics of the C1si makes it ideal for the high data acquisition rates necessary for live cell spectral imaging.
Ultimately the researcher can extend the life of cells under observation by reducing phototoxicity and photobleaching, while easily capturing dynamic and kinetic changes to the cells, says Nikon.
