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Super-Resolution Microscope N-STORM

Super-resolution microscope system which exceeds traditional diffraction limits by an order of magnitude and is capable of 3D image acquisition.

Super-Resolution microscope system offering ten times the resolution of conventional optical microscopes.
N-STORM is a super-resolution microscope system that combines “STochastic Optical Reconstruction Microscopy” technology (licensed from Harvard University) and Nikon’s Eclipse Ti research inverted microscope. The N-STORM super-resolution microscope provides dramatically enhanced resolution that is 10 times that of conventional optical microscopes.

Ten Times the Resolution of Conventional Optical Microscopes

N-STORM utilizes a high accuracy localization information for thousands of individual fluorophores present in a field of view to create breathtaking super-resolution images exhibiting spatial resolution of 20nm; ten times greater than conventional optical microscopes.

STORM-from-CO

3D Imaging with 50 nm Axial Resolution

In addition to lateral super-resolution, N-STORM utilizes proprietary methods to achieve a tenfold enhancement in axial resolution to 50nm, effectively providing 3D information at a nanoscopic scale

 

single-color3D

Single color 3D-STORM image of mitochondria in a BSC-1 cell labeled with Alexa405-Alexa647

Color encodes z-position information

Unique Multi-color Technology

Multi-color super-resolution imaging can be carried out using several dye strategies.

Unique tandem dye pairs (e.g. Alexa 405-Alexa 647, Cy2-Alexa 647, Cy3-Alexa 647) that combine “activator” and “reporter” probes allow highly accurate localization with no z shift.

Continuous Activation Imaging with a Single Dye

An alternative dye strategy is to use standard commercially available secondary antibodies for continuous activation imaging to easily gain critical insights into the localization of multiple proteins at the molecular level.

N-STORM_continuous_activation

Combination with Confocal Microscopy System A1R+

The N-STORM system is based around the Nikon Ti-E inverted microscope. This flexible platform allows combination with confocals such as the A1R+ for live-cell imaging, low magnification observation, photo-stimulation and 20nm-resolution imaging on one integrated system, under the control of the universal NIS-Elements platform.

 

 

N_STORM_A1R

 

Combination with Super-Resolution N-SIM

N_STORM_N-SIM

N-SIM and N-STORM can be combined on a single Nikon Ti-E inverted microscope to create the ultimate super-resolution imaging system. Using the N-SIM/N-STORM kit, switching between live cell super-resolution and 20nm-resolution imaging is possible without having to change the camera adapter.

 

Fit Overlapping Peaks Function

  • Identifies and fits more molecules per frame
  • Works for densely labeled samples
  • Increases reconstruction localization density
  • Can be used for 2D and 3D STORM data sets
  • Based on a published algorithm (Babcock et al., 2012. Optical Nanoscopy. 1:6)
FIT1

Original Data

FIT2

Detected Molecules with
Conventional Fitting

FIT3

Detected Molecules with
Fit Overlapping Peaks

351,560 mol.

FIT4

Without FOP

1,065,088 mol.

FIT5

With FOP

Objective for Super-resolution Imaging

The SR Apochromat TIRF 100x 1.49 N.A. objective lens is designed to provide the highest quality point spread function for N-STORM and N-SIM super-resolution imaging

 

SR_plan_apo_100

 

 

 

Super-Resolution Lateral (XY) ~20-30nm*
Axial (Z) ~50nm*
3D STORM Axial Range Per Single Data Set ±500nm (up to 20 localized planes)
Microscope Platform Ti-E TIRF with Perfect Focus System
Compatible Objectives  Plan Apo VC 100X 1.40 Oil
Apo TIRF 100X 1.49 Oil
Available Lasers, AOTF Modulated 405nm (20mW at the fiber tip)
457-514nm (70mW at the fiber tip)
561nm (70mW at the fiber tip)
647nm (125mW at the fiber tip)
Anti-Vibration Optical Table Required
Camera EM-CCD cameras iXon3 897, iXon Ultra 897 (Andor), and Evolve 512 Delta (Photometrics)**.  Higher frame-rates can be achieved with the iXon Ultra and Evolve Delta.
Fluorescence Probe Combination (Photo-switchable STORM probes)***

*Resolution depends on dye properties

**Camera compatibility depends on system configuration.

***Includes, but not limited to tandem-dye pairs

 

Quantitative analysis tools and correlative imaging applications for N-STORM

storm-note
DOWNLOAD: 348KB

Super-resolution techniques have had a significant impact on our understanding of biological processes at the molecular level. However, one of the challenges to their broad utilization has been our limited ability to quantitatively analyze super-resolution images of complex biological tissues. In this Application Note, we highlight recent work by Dudok et al. utilizing Nikon’s N-STORM system to develop new correlative imaging methods and quantitative analysis tools to study the mechanism of cannabinoid signaling in the brain.