Imaging Beyond the Diffraction Limit
Super-resolution. It's one of the most significant developments in biological imaging since the invention of the microscope. Super-high Resolution Microscopes, with capabilities that seemed impossible just a decade ago, have arrived on the scene, greatly extending the boundaries of cellular imaging. By breaking the diffraction limit and improving the resolution of the imaging sensor, we can suddenly observe the smallest of cellular structures with new levels of clarity down to almost the molecular level.
Nikon's Approach to Super-Resolution
Nikon has recently introduced two new systems, based on two highly effective super-resolution technologies developed by researchers at Harvard and UCSF, respectively.
STORM (STochastic Optical Reconstruction Microscopy)
STORM is a powerful technique that uses photo-switchable fluorescent probes to temporally separate the otherwise spatially overlapping images of individual molecules, allowing the construction of super resolution images. Using this concept, two- and three-dimensional, multicolor fluorescence images of molecular complexes, cells and tissues with a few tens of nanometers resolution can be achieved. This new form of fluorescence microscopy allows molecular interactions in cells and cell-cell interactions in tissues to be imaged at the nanometer scale.
Nikon's new N-STORM super-resolution system utilizes this groundbreaking technology. Integrated with an Eclipse Ti inverted microscope, N-STORM provides dramatically enhanced resolution that is 10 times or better than that of conventional optical microscopes and is capable of multi-spectral two-dimensional and three-dimensional nanoscopy, with lateral resolution to approximately 20nm and axial resolution to approximately 50nm. N-STORM extends the role of the optical microscope to near molecular level resolution.
Learn more
View STORM image gallery
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SIM (Structured Illumination Microscopy)
Structured Illumination Microscopy, or SIM, is a technique which takes advantage of moiré patterns, which are produced by overlaying one pattern with another. The sample under the lens is observed while it is illuminated by a special grid pattern of light. Several different light patterns are applied, and the resulting moiré patterns are captured each time by a digital camera. Computer software algorithms then extract the information in the moiré images and translate it into two- and three-dimensional, high-resolution reconstructions.
Nikon's N-SIM system produces up to two times the resolution of conventional optical microscopes by combining SIM technology licensed from UCSF and based on the world renowned Eclipse Ti research inverted microscope with Nikon’s legendary CFI Apo TIRF 100x oil objective lens (N.A. 1.49). N-SIM is incredibly fast, with temporal resolution up to 0.6 sec/frame.
Learn more
View SIM movie gallery
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Welcome to BioImaging Insights, a monthly newsletter aimed at keeping you abreast of the latest in microscope and digital imaging techniques and technology.
This month, our spotlight is on microscopy's latest breakthrough: Super-resolution. We detail the technology behind Nikon's newest SR products and provide links to key articles and research papers on the subject.
Resources
Video
Super-Resolution Named Nature Method of the Year
Nature Methods recognized super-resolution as the Method of the Year in 2008, touting its tremendous potential for understanding cellular biology.
Articles
Super-Resolution Microscopy: Breaking the Limits
Traces the emergence of super-resolution fluorescence microscopy as a powerful tool for biologists.
Source: Nature Methods
Super-Resolution Microscopy Captures Cells in Motion
Brief article on the Howard Hughes Medical Institute's role in the development of super-resolution. Includes live Structured Illumination Microscopy movies.
Source: HHMI Research News
Research Papers
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)
Outlines the development of STORM, a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores.
Source: Nature Methods, March, 2006
Authors: Michael J Rust, Mark Bates & Xiaowei Zhuang
Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy
Introduces SIM technology and how it produces images of strikingly increased clarity compared to both conventional and confocal microscopes.
Source: Journal of Microscopy, March, 2000
Author: M. G. L. Gustafsson
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