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- Structured illumination light sheet microscopy (SI-LSM) is an optical imaging technique used for achieving with high temporal and spatial resolution in all three dimensions. It combines the ability of light sheet microscopy to maintain spatial resolution throughout relatively thick samples with the higher axial and spatial resolution characteristic of structured illumination microscopy. SI-LSM can achieve lateral resolution below 100 nm in biological samples hundreds of micrometers thick. SI-LSM is most often used for fluorescent imaging of living biological samples, such as cell cultures. It is particularly useful for longitudinal studies, where high-rate imaging must be performed over long periods of time without damaging the sample. The two methods most used for fluorescent imaging of 3D samples – confocal microscopy and widefield microscopy – both have significant drawbacks for this type of application. In widefield microscopy, both in-focus light from the plane of interest as well as out-of-focus light from the rest of the sample is acquired together, creating the “missing cone problem” which makes high resolution imaging difficult. Although confocal microscopy largely solves this problem by using a pinhole to block unfocused light, this technique also inevitably blocks useful signal, which is particularly detrimental in fluorescent imaging when the signal is already very weak. In addition, both widefield and confocal microscopy illuminate the entirety of the sample throughout imaging, which leads to problems with photobleaching and phototoxicity in some samples. While light-field microscopy alone can address most of these issues, its achieved resolution is still fundamentally limited by the diffraction of light and it is unable to achieve super-resolution. SI-LSM works by using a patterned rather than uniform light sheet to illuminate a single plane of a volume being imaged. In this way, it maintains the many benefits of light-sheet microscopy while achieving the high resolution of structured illumination microscopy. (en)
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- Structured illumination light sheet microscopy (SI-LSM) is an optical imaging technique used for achieving with high temporal and spatial resolution in all three dimensions. It combines the ability of light sheet microscopy to maintain spatial resolution throughout relatively thick samples with the higher axial and spatial resolution characteristic of structured illumination microscopy. SI-LSM can achieve lateral resolution below 100 nm in biological samples hundreds of micrometers thick. (en)
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- Structured Illumination Light Sheet Microscopy (en)
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