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Nikon Crest DeepSIM Spinning Disk

Introduction:

Integrated spinning-disk confocal (Crest X-Light V3) microscopy with dual-cam acquisition and DeepSIM structured illumination microscopy (SIM) on a Nikon Ti2 platform. This system provides high frame rate spinning disc confocal imaging for a variety of samples, a broad excitation range and SIM super-resolution imaging with a lateral resolution of ~100 nm. It is the imaging platform for our current MERFISH initiative.

Main features and applications:

  • Fast and gentle imaging with spinning-disk confocal of standard or dim samples

  • Dual-cam and hardware-triggered imaging for “double” acquisition speed of multicolor samples

  • Super-resolution SIM imaging (~100 nm XY) of sub-cellular structures

  • Simultaneous multi-color imaging across visible to near-infrared excitation

  • Imaging platform for Multiplexed Error-Robust Fluorescence in situ Hybridization (MERFISH) at DaMBIC

 

Specifications

Microscope body:
Nikon Eclipse Ti-2 inverted microscope with Perfect Focus System (PFS).

Compatible sample holders

  • Standard ~25 x 75 mm glass slides + chambered glass slides

  • Culture dishes w. diameter: ~30 – 40 mm

Scan mechanism, spinning disk:

Crest X-Light V3) with 2 disk options:

  • Standard 50 µm pinhole with 250 µm spacing. Perfect for cell monolayers or thin sections (2-20 µm). Confocality (axial resolution) ~40 % better than widefield.

  • Slit disc with pinhole size of 70 µm. Suitable for very dim samples as they allow more light through the pinhole at the expenses of confocality. Confocality (axial resolution) ~20-25 % better than widefield.

Objectives:

Nikon objectives from low to high magnification (2x – 60x), including high-NA oil/silicon immersion.

  • 2× / NA 0.06 Plan UW

  • 10× / NA 0.45 Plan Apo Lambda D

  • 20× / NA 0.80 Plan Apo Lambda D

  • 25× / NA 1.05 Plan Apo Lambda S Sil

  • 40× / NA 1.25 Plan Apo Lambda S Sil60× / NA 1.42 Plan Apo Lambda D Oil

 

Light Sources / filters:

  • Lumencor Celesta Quattro 7ch with Despeckler, lines 405 nm, 446 nm, 476 nm, 518 nm, 545 nm, 637 nm and 748 nm.

  • Various dual- and pentaband excitation and emission filter options are available in the spinning disk unit.

 
Modules:

  • CrestOptics X-Light V3 spinning-disk (25 mm FOV, disk up to 15,000 RPM)

  • DeepSIM module (SIM FOV examples: 66 × 66 µm @ 100×; 333 × 333 µm @ 20×)

 
Camera(s):

2x Teledyne Photometrics Kinetix back-illuminated sCMOS camera with large fields of view and 95% quantum efficiency.

  • FOV: 20.8 x 20.8 mm

  • Speed: 500 fps

  • Pixel size: 6.5 µm

  • QE: 95% (at ~570 nm)

  • Read noise: 0.7 e-

 
Stage & Environment:

  • Motorized stage and objective for automated multi-point imaging

  • Nikon Perfect Focus System (PFS) for hardware-based autofocus

  • Prior piezo Z-stage for fast/precise imaging in Z

 

Software: NIS-Elements 6.10 with ND acquisition and JOBs.

Last Updated 22.10.2025