Funding
Nikon N-SIM S microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 496847469.
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 |
Nikon Imaging software NIS Elements AR (Advanced Research). NIS-A Bundle JOBS 2.0.
