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The spinning disk confocal microscope, fi rst popularized for its reliability and


speed in live-cell imaging, can be adapted for super-resolution performance.


Point-scanning SIM Most super-resolution techniques rely on software to process image data and provide accurate estimates of the location of subdiff raction- limit structures and dynamics. For mathematical simplicity, most current implementations of SIM use sinusoidal illumination patterns and compu- tational image reconstruction.


However, systems that perform optically what otherwise would be done computationally have advantages in speed and simplicity. Mats Gustafsson, the inventor of SIM technology, asserted that the optical microscope could produce better images at far-higher resolutions by applying structured or patterned excitation. Today, new point-scanning and spinning-disk systems combined with modern detectors, improved optics and insightful mathematical models make super resolution more accessible than ever, just as Dr. Gustafsson predicted.


The spinning disk confocal microscope, fi rst popularized for its reliabil- ity and speed in live-cell imaging, can be adapted for super-resolution performance. For instance, the Olympus SD-OSR (Olympus America, Waltham, MA) pairs a high-speed spinning-disk microscope system with a high-performance silicone oil objective for an ideal optical match to living samples , to deliver image resolution up to twice that of a widefi eld microscope without localization (Figure 1). Resolution-limited excitation, reduced-noise image acquisition and high-performance optics combine with SIM image formation to deliver 110–130 nm resolution at up to three frames per second (Figure 2).


Conclusion Biological microscopy involves a continuous quest to image what es-


sentially cannot be seen—the subtle and fast-changing processes of life. Super resolution provides another tool that researchers can use to learn more about the structures and dynamics of life. Each technique has its strengths and is ideally suited for a certain type of imaging, but also has factors that limit its usefulness.


The next step in super resolution may allow researchers to image faster- moving, ever-smaller structures for longer periods, even deeper within tissue. However, there will always be an object at the edge of resolution, and science will dwell on the phenomena that lie just beyond.


Russell Ulbrich is product manager, Software and Advanced Imaging Systems, Olympus America Inc., 48 Woerd Ave., Waltham, Mass. 02453, U.S.A.; tel.: 781-419-4686; e-mail: russell.ulbrich@olympus.com; www. olympusamerica.com


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