Nikon’s A1R MP multiphoton confocal microscope is a unique multiphoton imaging system featuring a fast, high resolution galvanometer scanner and an ultra-high speed resonant scanner that is capable of frame rates from 30 fps at 512 x 512 pixels to as fast as 420 fps in band scan mode. New 4 channel non-descanned detectors with higher sensitivity, reduced dark current and broad spectral range allow for real time unmixing of closely spaced probes for accurate and high-contrast spectral imaging. This is especially important in multiphoton microscopy because of the overlap of emission spectra of probes and autofluorescence, which is often unavoidable when using a single laser line.
Nikon\'s exclusive resonant scanner is capable of imaging a wide area at a much higher speed than a non-resonant scanner, making it possible for 420-fps imaging— the world\'s fastest using point scanning technology. The NDD for multiphoton microscopy makes it possible to image fast and deep through the thickest specimens. Nikon\'s optical pixel clock system allows more stable and more evenly illuminated imaging— even at high speeds.
Nikon\'s newly developed 4-channel NDD for multiphoton microscopy allows imaging deep into a specimen. A detector* with very high sensitivity and a wider sensitive area than conventional models is placed close to the back aperture of the objective where image formation of the specimen takes place. This configuration improves the detection efficiency for scattered fluorescence, and enables the capture of signals from deep within a living organism more clearly and more reliably with higher S/N and less aberrations.
In addition to spectral detection and unmixing by the 32-channel detector, unmixing by the 4-channel NDD for multiphoton microscopy is also realized. Images captured with a resonant scanner can be unmixed, so clear and high-contrast images of areas deep within a thick specimen can be captured in ultrahigh speed.
When the multiphoton laser wavelength or group velocity dispersion precompensation is changed, the multiphoton laser beam positional pointing may drift, resulting in uneven intensity of images, or a slight misalignment of fluorescence images that are produced by a visible light laser and a multiphoton laser.
Since the multiphoton laser is invisible to the eye, the laser beam alignment has been difficult and potentially dangerous for users, especially at wavelengths longer than 800 nm. The newly developed auto laser alignment function of the Nikon A1R MP+ automatically corrects multiphoton laser misalignments with a single click.
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