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

Description:

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.
 

Specifications:

• Standalone or correlative confocal, SIM and STORM imaging
• Eclipse Ti fully motorised inverted microscope body
• Adaptive auto-focusing system (Nikon Perfect Focus System)
• High-precision motorised stage with encoder
• Tile scanning and image stitching to produce high-resolution overview image of large tissue sections
• Motorised condenser, objective and fluorescence filter turrets
• Transmitted light detector for bright-field and differential interference contrast (DIC) imaging
• Multi-wavelength LED fibre optic illumination system (CoolLED pE-300^white) with spectral coverage from ultraviolet to red region
• Environmental chamber with temperature and CO₂ concentration control (Tokai Hit INU series)
• Objectives (details):
• CFI Plan Apo 10×/0.45 Dry
• CFI Plan Apo 20×/0.75 Dry
• CFI Plan Apo 40×/0.95 Dry
• CFI Plan Apo 40×/1.25 Silicone Oil
• CFI Plan Apo 60×/1.40 Oil
• CFI SR Plan Apo 60×/1.27 Water (compatible with SIM)
• CFI SR Apo TIRF 100×/1.49 Oil (compatible with SIM)
• CFI HP Apo TIRF 100×/1.49 Oil (compatible with STORM)
• CFI SR HP Plan Apo 100×/1.35 Silicone Oil (compatible with SIM & STORM)
• Excitation lasers:
• 405 nm
• 488 nm
• 445 nm (confocal only)
• 561 nm
• 647 nm
• N-SIM module:
• XY resolution: 115 nm
• Z resolution: 250 nm
• Imaging modes: 2D-SIM, 3D-SIM, TIRF-SIM
• N-STORM module:
• XY resolution: 20 nm
• Z resolution: 50 nm
• Imaging modes: 2D-STORM, 3D-STORM
• A1 HD25 confocal module:
• Large (i.e., 25 mm) field-of-view
• Dual-axis galvano scanner
• Maximum image size: 4096 × 4096 pixels
• Scan modes: XY, XT, XZ, XY rotation and free line
• Hexagonal, continuously adjustable pinhole (size ranges from 12 to 256 µm)
• 4-channel detector: 2 GaAsP and 2 multialkali PMT modules
• Detection range: 400 to 750 nm

Location: Y708b

Hourly rate: $198 (internal users); $396 (collaborators); $990 (external users)

Discount for long-duration bookings:

• Applicable to consecutive bookings of 8 hours or longer
• A 50% discount will be applied to any segments that fall within non-office hours (i.e., from 6 pm to 9 am on weekdays, and the whole days on Saturdays, Sundays and public holidays)
• Price adjustments, if any, will be made after the completion of the imaging session
• The ULS reserves the right to interpret these terms

Consumables:

• Marienfeld no. 1.5H 18 mm round coverglass ($148; 100 pcs)
• MatTek 35 mm no. 1.5 glass bottom culture dish ($190; 10 pcs)
• Nunc Lab-Tek II no. 1.5 8-well chambered coverglass ($435; 8 pcs)
• ProLong Diamond antifade mountant ($372; 2 mL)
• ProLong Live antifade reagent ($333; 1 mL)
• EMS 16% paraformaldehyde ($116; 10 mL)

Staff-in-charge: Dr Michael Yuen and Dr Alan Leung

Training: please contact Dr Michael Yuen (email: michael.yf.yuen@polyu.edu.hk / Tel.: 3400 8788) or Dr Alan Leung (email: alan.sl.leung@polyu.edu.hk / Tel.: 3400 8984) for arrangement.