ICCE Equipment - Nikon N-SIM S

© UDE/Bettina Engel-Albustin

Superresolution SystemsNikon N-SIM S

Room: T03 R01 D66

Main Applications

  • multi-color 2D/3D-SIM
  • mulit-color TIRF-SIM
  • TIRF
  • photostimulation (FRAP, photoconversion, optogenetic approaches)
  • fast multi-dimensional live cell + fixed specimen imaging
  • multi-position acquisition
  • simultaneous 2-color imaging (dual cam)

 

Funding

Nikon N-SIM S microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 496847469.

 

Technical Specifications

Microscope and Accessories

  • inverted Nikon Ti2-E
  • Perfect Focus System PFS4 (hardware autofocus system)
  • motorized xy-stage
  • N-SIM S illuminator
  • Nikon Ti LAPP system with modules:
    • epi-fluorescence module
    • motorized TIRF-E SIM module
    • DMD module (photostimulation device)
  • TokaiHit stage top incubation system with control device (temperature & CO2 control for live-cell imaging)
  • TokaiHit drug delivery system KSX-Type2

Cameras and DualCam

  • 2 x: Hamamatsu Orca FusionBT bck-thinned sCMOS camera, 2304 x 2304 pixels imaging array, 6.5 x 6.5 µm pixel size, 16-bit digitization, up to 31.6 fps at full frame/fast scan
  • Hamamatsu W-VIEW GEMINI-2C

Light sources and lasers

LED transmitted light module

Lumencor Spectra III for epi and DMD

Line Excitation Power
DAPI 390/22 nm 500 mW
CFP 440/20 nm 500 mW
GFP/FITC 475/28 nm 500 mW
YFP 510/25 nm 400 mW
Cy3 555/28 nm 500 mW
mCherry 575/25 nm 500 mW
Cy5 635/22 nm 500 mW
Cy7 747/11 nm 500 mW

Lumencor ZIVA for 2D- and 3D-SIM (solid state lasers)

Line Excitation Power
DAPI 405 nm 20 mW
CFP 446 nm 100 mW
GFP/FITC 476 nm 100 mW
YFP 518 nm 100 mW
Cy3 545 nm 100 mW
Cy5 637 nm 100 mW

Nikon LUDH for TIRF and TIRF-SIM (solid state lasers)

Line Excitation Power (fiber 1/ fiber 2)
DAPI 405 nm 40 mW / 50 mW
CFP 445 nm 40 mW / 50 mW
GFP/FITC 488 nm 80 mW / 100mW
YFP 515 nm 40 mW / 50 mW
Cy3 561 nm 70 mW / 70 mW
Cy5 640 nm 112 mW / 140 mW

Fluorescence filters & dichroics

 

name Fluorochromes excitation [nm] dichroic [nm] emission [nm]
MXR00249 DAPI/FITC/TRITC/Cy5/Cy7

352-404

461-487

543-566

626-644

721-749

414-450

499-530

580-611

661-701

768-849

420-460

510-531

589-623

677-711

768-849
MXR00241 CAP/YFP/mcherry

426-450

497-519

567-588

464-486

532-554

603-800

464-586

532-554

603-800
YFP YFP 490-510 525+ 520+
Continuous STORM  

387-417

483-494

557-570

636-661

420-481

497-553

575-628

667-792

422-478

502-549

587-625

674-786
AHF-TIRF    

405-445

510-520

555-570

455-500

525-550

575-673

Objectives

name

magnification

 NA immersion
CFI Plan Apochromat Lambda D 20x 20x 0.8 dry
CFI SR Plan Apochromat 60x IR WI 60x 1.27 water
CFI Apochromat TIRF 60x 60x 1.49 oil
CFI SR HP Apochromat TIRF 100xAC 100x 1.49 oil

 

Software

Nikon Imaging software NIS Elements AR (Advanced Research). NIS-A Bundle JOBS 2.0.