Achieving a resolution 10 times greater than a conventional optical microscope enables molecular level understanding
STochastic Optical Reconstruction Microscopy (STORM) reconstructs a super-resolution fluorescent image by combining precise localization information for individual fluorophores in complex fluorescent microscope specimens. N-STORM takes advantage of Nikon’s powerful Ti-E inverted microscope and applies high-accuracy, multi-color localization and reconstruction in three dimensions (xyz) to enable super-resolution imaging at tenfold the resolution of conventional optical microscopes (up to approximately 20 nm in xy)
This powerful technology enables the visualization of molecular interactions at the nanoscopic level, opening up new worlds of scientific understanding.
N-STORM offers 20 nm lateral resolution, a tenfold improvement over conventional optical microscopes
N-STORM utilizes high accuracy localization information for thousands of individual fluorophores present in a field of view to create breathtaking “super-resolution” images, exhibiting spatial resolution that is 10 times greater than conventional optical microscopes.
• Single color STORM image of a clathrin-coated pit in a mammalian cell labeled with Cy3-Alexa647
Objective: CFI Apochromat TIRF 100x oil (NA 1.49)
N-STORM also offers more than tenfold improvement in axial resolution (up to approximately 50 nm)
In addition to lateral super-resolution, N-STORM utilizes proprietary methods to achieve a tenfold enhancement in axial resolution, effectively providing 3D information at a nanoscopic scale.
Multi-color imaging using various fluorescent probes
Multi-color super-resolution imaging can be carried out using either dedicated secondary antibodies that conjugate tandem dye pairs by combining “activator” and “reporter” probes, or standard secondary antibodies that are commercially available for continuous activation imaging. This flexibility allows users to easily gain critical insights into the localization and interaction properties of multiple proteins at the molecular level.
• Dual color STORM image of microtubule (Alexa405-Alexa647) and mitochondria (Cy3-Alexa647) in a mammalian cell.
Objective: CFI Plan Apochromat VC 100x oil (1.40)
The principle of STochastic Optical Reconstruction Microscopy
STochastic Optical Reconstruction Microscopy (STORM) reconstructs a super-resolution image by combining the high-accuracy localization information of individual fluorophores in three dimensions and multiple colors
N-STORM uses stochastic activation of relatively small numbers of fluorophores using very low-intensity light. This random stochastic “activation” of fluorophores allows temporal separation of individual molecules, enabling high precision Gaussian fitting of each fluorophore image in XY. By utilizing special 3D-STORM optics, N-STORM can also localize individual molecules along the Z-axis with high precision. Computationally combining molecular coordinates in three dimensions results in super-resolution 3D images of the nanoscopic world.
Dedicated tandem-dye pairs for highest localization accuracy
N-STORM uses dedicated secondary antibodies that conjugate fluorescent dye pairs containing an "activator" (relatively short wavelength excitation) and a "reporter" (relatively long wavelength excitation), which enables various color combinations, facilitating true multi-channel super resolution. N-STORM can also be carried out using conventional single-dye conjugated antibodies for continuous activation imaging.
• A dye for N-STORM consists of a shorter-wavelength dye for activation and a longer-wavelength dye for image capturing. Creation of three-color super-resolution images is possible with multiple dye-pairs.
Objectives for super-resolution microscopes
The SR (super resolution) objectives have been designed to provide superb optical performance with Nikon’s super-resolution microscopes.
The adjustment and inspection of lenses using wavefront aberration measurement have been applied to yield optical performances with the lowest possible asymmetric aberration.
CFI SR Apochromat TIRF 100x oil
• *CFI Apochromat TIRF 100× oil and CFI Plan Apochromat VC 100x oil can also be used.
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