Nikon N-SIM/N-STORM/A1 Super-resolution/Confocal Microscope

The best resolution of conventional light microscopy is limited to ~200 nm due to the diffraction of light. Super-resolution microscopy overcomes this diffraction limit and allows scientists to observe and study subcellular or even molecular processes in greater detail. This opens up new avenues in the research areas of cell and molecular biology, neuroscience and microbiology, etc.

In structured illumination microscopy (SIM), the sample is illuminated with a series of highly structured excitation light (e.g., a pattern of parallel lines). This structured illumination is able to produce images that carry high frequency information that lies beyond the diffraction limit. The encoded information can subsequently be reconstructed using special algorithms as super-resolution images with a lateral resolution of up to 115 nm.
Stochastic optical reconstruction microscopy (STORM) is a widely-used single-molecule localisation microscopy technique. In STORM, the sample needs to be densely labelled with photoswitchable fluorescent probes. During image acquisition, the activation laser stochastically activates a sparse set of probes, which are then imaged with the readout laser. The precise location of the point-spread functions of individual molecules are determined (based on photon output) before the probes switch back to the dark state. This process is repeated for thousands of times to obtain enough localisation information to reconstruct a super-resolution image with a lateral resolution of up to 20 nm.
In addition to serving as a standalone confocal or super-resolution microscope, the Nikon N-SIM/N-STORM/A1 system also allows researchers to perform correlative confocal-super-resolution imaging. In the same field-of-view (FOV), researchers can choose to use confocal for certain fluorescence channels, and super-resolution for the remaining ones. The final confocal images can then be overlaid with the corresponding super-resolution images in the same FOV. This is particularly useful when one needs to perform multi-colour imaging, and when not all fluorescent labels are compatible with super-resolution microscopy. Alternatively, researchers can choose to use confocal to scan a larger FOV to provide an overview of the sample, and use super-resolution to image a selected area of interest to examine the area in greater detail.