Achieving 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 Ti2-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 is fully improved as N-STORM 4.0 that allows high-speed STORM imaging of intracellular dynamics and imaging of large field of view.
Tenfold improvement of lateral resolution up to 20nm
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.
Human cervical cancer cells (HeLa S3) labeled with Alexa Fluor® 647 (NUP153) and ATTO 488 (TPR)
Photos courtesy of: Dr. Michael W. Davidson, National High Magnetic Field Laboratory, Florida State University
Tenfold improvement of axial resolution up to 50 nm
In addition to lateral super-resolution, N-STORM utilizes proprietary methods to achieve a tenfold enhancement in axial resolution over conventional optical microscopes, and effectively provide 3D information at the nanoscale.
3D-Stack function allows multiple 3D STORM images in different Z positions to be captured and merged into one image to create thicker STORM image.
3D-Stack STORM image of tubulin of BSC-1 cell labeled with Alexa Fluor® 647 with image depth of 4µm.
Dynamic super resolution imaging at the nanoscale level
Image acquisition speed has been significantly improved, increasing from minutes to seconds* for a single shot, due to newly developed optics and illumination systems optimized for the sCMOS camera, which is capable of approximately 10 times faster image acquisition than before. Thanks to this improvement, it is now possible to acquire dynamics of living specimens at a resolution 10 times greater than that of conventional optical microscopes.
- *Using high-speed mode (20µm x 20µm imaging area)
Imaging speed: 350 fps
30 min time-lapse imaging with 1 min interval
Multi-color imaging capability
Multi-color super-resolution imaging can be carried out using either dedicated secondary antibodies that are conjugating tandem dye pairs 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 (Alexa Fluor® 405-Alexa Fluor® 647) and mitochondria (Cy3-Alexa Fluor® 647) in a mammalian cell.
Objective: CFI Plan Apochromat VC 100X Oil (NA 1.40)
High definition, high density images
The newly developed illumination magnifying lens, improved laser excitation efficiency, and increased image acquisition rate successfully enhance the density of molecules per unit area and provide much clearer images with high molecule counts.
Left: Improved image quality, Right: Before improvement
Scale bar: 5µm
Super-resolution image quality is significantly improved in the same imaging time.
Sample: Tubulin of BSC-1 cell labeled with Alexa Fluor® 647, acquisition time: 20 seconds
Large image acquisition area
Intermediate zoom lenses in the imaging system have been newly developed and optimized for a wide field of view.
The wide-view mode is achieved at 80 µm x 80 µm, which is an imaging area 4 times wider than before.
Left: 4 times wider imaging area, 80 µm x 80 µm (wide-view mode)
Right: Imaging area of conventional model, 40 µm x 40 µm
Sample: Mitochondria TOM20 conjugated with Alexa Fluor® 647
Objectives optimized for N-STORM imaging
The HP objective is compatible with the high power lasers required for the rapid blinking of fluorophores. Due to improved axial chromatic aberration correction, higher precision than ever before is possible in 3D multi-color fluorescence imaging.
The SR HP objective provides greater image quality in 3D STORM images by achieving superior super-resolution imaging performance.
In addition, the CFI SR HP Plan Apochromat Lambda S 100XC Sil employs silicone immersion oil (ne≒1.40) to reduce spherical aberrations caused by difference in the refraction indexes of immersions and living cell specimens (ne≒1.38), enabling accurate fluorescence observation from the surface to the deep areas of specimens.