Introduction to Super Resolution Microscope

 Introduction to Super Resolution Microscope N-SIM








 There is a universal desire among life science researchers to observe tissues and cells more clearly. Optical microscopes are essential for this purpose. However, if multiple objects such as protein molecules cluster at distances of less than 200 nm apart, conventional optical microscopes cannot identify them as single objects. In this case, other instrumentation such as electron microscopes must be used. Nikon's super resolution fluorescence microscopy technology greatly exceeds the resolution limits of conventional optical microscopes, making it possible to view microstructures and nanostructures of living cells with molecular-scale resolution. In addition, it can continuously capture super resolution images at temporal resolution of 0.6 sec/frame, effective for live-cell imaging. Recently, the Core Instrument Center at NHRI has purchased this system. 

Key Features
1.    Nearly double the resolution of conventional optical microscopes
The N-SIM super resolution microscope utilizes Nikon's innovative new approach to “Structured Illumination Microscopy” technology. Multiple phases and orientations of structured illumination are captured, and the displaced “super resolution” information is extracted from moiré fringe information. This information is combined mathematically in "Fourier" or aperture space then transformed back into image space creating an image at double the conventional resolution limit.
By pairing this powerful technology with Nikon's renowned CFI Apo TIRF 100x oil objective lens (NA 1.49), N-SIM nearly doubles (to approx. 85nm) the spatial resolution of conventional fluorescence and confocal microscopes, and enables detailed visualization of the minute intracellular structures and their interactive functions.
2.    Time resolution of 0.6 sec/frame
N-SIM provides the fastest imaging capability in the industry, with a time resolution of 0.6 sec/frame in TIRF-SIM/2D-SIM mode; imaging of up to approx. 1 sec/frame is possible with 3D-SIM mode, enabling the study of dynamic interactions in living cells.
3.    TIRF-SIM imaging technique
The newly developed TIRF-SIM illumination technique enables Total Internal Reflection Fluorescence (TIRF) observation with higher resolution than conventional TIRF microscopes and gives more detailed structural information near cell membrane.
4.    3D-SIM imaging technique
New 3D-SIM illumination technique has the capability of optical sectioning of specimens at 300nm resolution in cells and tissues of up to 20µm thickness, enabling the visualization of more detailed cell spatial structures.
The super resolution microscope could be applied in the fields of:
-       Neuroscience (Alzheimer’s disease)
-       Immunology (Regenerative medicines)
-       Developmental Biology (iPS cell)
-       Cell biological study (Gene manipulation)
-       Physiology (Molecularly-targeted drug)
-       Genetics (Tailor-made medicine)
This N-SIM super resolution microscope is located in the NHRI Optical Biology Core Laboratory and is full-equipped with 3 imaging modes (2D-SIM, TIRF-SIM, and 3D-SIM) as described above and the 488 nm and 561 nm lasers for fluorescence image acquisition.
<Contents and Figures are provided by Nikon Corporation and NHRI Core Instrument Center>



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