A1R Resonant Scanning Multispectral Confocal Microscope
The Nikon A1R laser scanning confocal microscope provides several important features:
1. Resonant scanner
The Nikon A1R is equipped with a high-speed resonant scanner that will allow users to scan their sample at 7.8kHz. This scan speed translates into high-speed imaging at 420 fps (512 x 32 pixels). Since the field of view is perfectly aligned with that generated by the conventional scanner, users can easily follow very fast biological process even when combined with photobleaching experiments, for example. This fast scanning feature is also tailored for imaging rapid signaling events such as calcium waves.
2. Image stitching
Nikon A1 is capable of combining image-stiching in three-dimension. This will allow users to create a montaged Z-stacks of multiple fields of view at high resolution. This is particularly powerful if high resolution and/or 3-D image of a large section of a specimen is needed. In addition,
The A1R system is fitted on Nikon Ti PerfectFocus microscope stand. This robust focal drift compensation mechanism thus allows users to maintain the desired focal plane for long-tem live cell imaging, even combining drastic temperature changes into their experiments (such as heat-shock) if necessary without the worry of thermal instability and focal plane shift.
4. Excitation lasers
The Nikon A1R is equipped with multiple excitation wavelengths (405nm, 458nm, 488nm, 514nm, 561nm, and 633nm). Combined with its mulsipectral and resonant scanning capabilities, this system will allow user to perform fast multiplexed experiments, as well as specialized live cell imaging experiments such as FRET, photoactivation and photoconversion with great ease.
5. Multispectral unmxing
The Nikon A1R is also fitted with multispectral detector that allows users to performing emisson fingerprinting. The ability to separate emission based on spectral properties rather than through band-pass filtering significantly enhances the capability of the system to distinguish highly overlapped spectra, and therefore greatly increases the number of fluorophores that can be simultaneously used in a given specimen. In addition, it will also allow users to very specifically eliminate tissue autofluorescence without compromising the intensity of the actual fluorescence staining.
6. Nikon Elements software
The Nikon Elements software not only provides intuitive user interface to seanlessly perform many types of experiments, it also has easy-to-use 3D rendering and morphometric segmenting capabilities. As shown below, users can generate 3D rendered videos and segment irregularly shaped structures at-will with the Elements software. The Nikon Imaging Center now has three full-version, stand-alone Elements software packages in various work stations, enhancing the general availability of this software to users who wants to perform post-acquisition analysis. Free version of the Elements Viewer can also be downloaded from the Nikon website.
The Elements software offers intuitive 3-D rendering capabilities that allows users to not only rotate the Z-stack at-will, but also peel back any fluorescent channel to reveal the structure underneath.
The Nikon Elements software also offers easy-to-define image segmentation functions for its user to effortlessly highlight objects for image analysis.
This instrument is purchased mainly with fund from the Northwestern University Office for Research, and in part with the support from the Feinberg School of Medicine.