Super-Resolution Microscope N-SIM

Structured illumination super-resolution microscope delivering twice the resolution of traditional diffraction limited microscopes.

Super-resolution microscope visualizing cellular structures and molecular activity at resolutions never before achieved by conventional light microscopy.
Nikon’s N-SIM microscopy system can produce twice the resolution of conventional optical microscopes. Using an innovative approach based on Structured Illumination Microscopy technology licensed from UCSF, the N-SIM enables detailed visualization of minute intracellular structures and their functions.

Twice the Resolution of Conventional Optical Microscopes

N-SIM doubles the resolution of conventional optical microscopes (~115nm in 3D SIM mode*) by combining “Structured Illumination Microscopy” technology licensed from UCSF with Nikon’s renowned CFI Apo TIRF 100x oil objective lens (N.A. 1.49).

* resolution will depend on laser wavelength and imaging mode.

Macrophages (J774 cells expressing mVenus-SNAP23) phagocytosing opsonized beads that were incubated with Alexa555 labeled secondary antibodies after fixation.

The beads without red signals are in phagosomes containing mVenus-SNAP23.

Photographed with the cooperation of: Drs. Chie Sakurai, Kiyotaka Hatsuzawa and Ikuo Wada, Fukushima Medical University School of Medicine.

Dedicated Super-resolution Objectives

The SR (super-resolution) objectives have been designed for new applications that break the diffraction barrier.

The most recent optical designs using wavefront aberration measurement have been applied to yield superb optical performances with the lowest asymmetric aberration.

Super-Resolution Objectives

Time Resolution of 0.6 sec/frame, the Fastest in the Industry

N-SIM provides the fastest imaging capability in the industry, with a time resolution of 0.6 sec/frame, effective for live-cell imaging.

N_SIM_mitochondria

Live-cell N-SIM imaging of mitochondria labeled with Mito-Tracker red.

Live-cell imaging with N-SIM reveals dynamics of mitochondria at twice the spatial resolution. Cristae in mitochondria are also clearly observed.
Mode: Slice 3D-SIM mode
Objective: CFI Apochromat TIRF 100x oil (NA 1.49)
Image capturing interval: approximately 1 sec. (movie)

Live Cell Super-Resolution Imaging

The combination of fast image capture and the availability of stage top incubation allows live cell imaging at twice the resolution of conventional microscopes.

Bacillus subtilis bacterium stained with membrane dye Nile Red (red), and expressing the cell division protein DivIVA fused to GFP (green).
N-SIM enables accurate localization of the protein during division.
Photos courtesy of: Drs Henrik Strahl and Leendert Hamoen, Centre for Bacterial Cell Biology, Newcastle University

Multiple SIM Modes

3D-SIM

The 3D-SIM illumination technique improves axial resolution to 269 nm and has the capability of optical sectioning of specimens, enabling the visualization of more detailed cell structures at higher spacial resolutions.

Luminal surface of the organ of Corti at postnatal day 1. (Mouse)
Green: F-actin, red: acetylated-tubulin
Photographed with the cooperation of: Drs. Kanoko Kominami, Hideru Togashi, and Yoshimi Takai, Division of
Molecular and Cellular Biology, Kobe University Graduate School of Medicine/Faculty of Medicine

TIRF-SIM/2D-SIM mode

This mode captures super-resolution 2D images at high speed with incredible contrast. TIRF-SIM mode takes advantage of Total Internal Reflection Fluorescence observation at double the resolution as compared to conventional TIRF microscopes, facilitating a greater understanding of molecular interactions at the cell surface.

Conventional TIRF image

TIRF-SIM image

Plasma membrane of B16 melanoma cell labeled with YFP
Objective: CFI Apochromat TIRF 100x oil (NA 1.49)
Photographed with the cooperation of: Dr. Yasushi Okada, Laboratory for Cell Polarity Regulation, Quantitative Biology Center, RIKEN

5 Laser Multi-color Super-resolution Imaging

The Nikon N-SIM system allows multi-channel imaging with up to 5 lasers enabling the study of dynamic interactions of multiple proteins of interest at the molecular level.

Human U2OS cell during mitosis metaphase
The cell is labeled green (kinetochore protein CENP-B), red (alpha-tubulin) and blue (DNA).
Photo courtesy of: Dr. Alexey Khodjakov, Wadsworth Center, Albany NY

Simultaneous Two-wavelength Super-resolution Imaging

By attaching two EMCCD cameras to the microscope with the optional Two Camera Imaging Adapter*, simultaneous two-wavelength super-resolution imaging with excitation of 488 nm and 561 nm is possible.

* Andor Technology Ltd.

Two Camera Imaging Adapter (for N-SIM)

* The actual product may differ slightly in design.

Super-Resolution	Lateral (XY): ~85-110nm (dependant on wavelength and optics)* Axial (Z): ~300nm (dependant on wavelength and optics)* 3D Axial Range: up to 20μm
Image acquisition time	Up to 0.6 sec/frame (TIRF-SIM/2D-SIM)Up to 1 sec/frame (Slice 3D-SIM) (needs more 1-2 sec. for calculation)
Imaging mode	TIRF-SIM2D-SIM Slice 3D-SIM Stack 3D-SIM
Multi-color imaging	Up to 5 colors
Compatible Laser	Standard: 488nm, 561nmOption: 405nm, 458nm, 514nm, 532nm, 640nm Laser combination: 405 nm/488 nm/514 nm/532 nm/561 nm, 405 nm/488 nm/514 nm/561 nm/640 nm, 458 nm/488 nm/514 nm/532 nm/561 nm, 458 nm/488 nm/514 nm/561 nm/640 nm
Compatible microscope	Motorized inverted microscope ECLIPSE Ti-EPerfect Focus System Motorized XY stage with encoders Piezo Z stage
Compatible objective	CFI SR Apochromat TIRF 100×oil (NA1.49)CFI Apochromat TIRF 100×oil (NA1.49) CFI SR Plan Apochromat IR 60×WI (NA1.27) CFI Plan Apochromat IR 60×WI (NA1.27)
Camera	EM CCD camera iXon3 DU-897E (Andor Technology Ltd.)
Software	NIS-Elements Ar/NIS-Elements C (for Confocal Microscope A1+/A1R+)Both require optional module software NIS-A N-SIM Analysis
Operating conditions	20°C to 28°C ( ± 0.5 oC)