Nikon Eclipse Ti2 - Widefield, TIRF, Spinning disk confocal
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This instrument is part of the NORBRAIN nanoscopy facility and is organized under MIC. The microscope was upgraded Sept 2025 with a CrestOptics V3 Spinning disk confocal module.
The Nikon Eclipse Ti2 is a versatile microscope and on this system the applications can be divided into two cathegories:
(I) Common widefield/brightfield microscopy applications and spinning disk confocal. Supported by Endy & Hege (MIC). Please find a user-guide below under Documents.
- Widefield epifluorescent microscopy - 4 color multiplexing
- Phase-contrast, brightfield microscopy w/color camera
- Timelapse (fast, long term, multi-positions)
- Stitching abilities -> large images
- Total internal reflection microscopy (TIRF) - 4 color multiplexing
- Spinning disk confocal - 4 color multiplexing
(II) Single molecule localization microscopy (SMLM) applications (see useful Nature review below under Documents). Supported by Hongyu Zhang (Bramham-lab).
- Single partical tracking (spt-PALM)
- PALM (Photo Activated Localization Microscopy)
- DNA-Paint
Technical highlights:
Cameras: Monochrome Prime BSI Express Scientific CMOS and color DS-Fi3 CMOS.
Light sources:
Widefield fluorescence imaging: CoolLED pE-300 Ultra MB
CrestOptics X-Light V3 spinning disk confocal: Lumencor Celesta Quattro, 4 channels; 405 nm, 477 nm, 546 nm and 640 nm
For TIRF and activation/bleaching experiments: LUN-F laser unit with 405 nm (70 nW), 488 nm (70 mW), 561 nm(60 mW) and 640 nm (50 mW) lasers.
Live cell incubator: OkoLab stage top heating incubator with CO2 control.
Objectives:
Magnification | Objective and correction options | NA | WD (mm) | Contrast method and application |
10 x Air | Plan Fluor | 0,30 | 15,2 | Phase contrast (Ph1) |
20 x Air | Plan Fluor (coverslip correction) | 0,45 | 7,4 | Phase contrast (Ph1) |
20 x Air | Plan Apochromat lambda | 0,75 | 1,0 | Fluorescence |
40 x Air | Plan Apochromat lambda (coverslip correction) | 0,95 | 0,21 | Phase contrast (Ph2) / fluorescence |
60 x Oil | CFI Apo TIRF (coverslip and temperature correction) | 1,49 | 0,12 | TIRF / fluorescence |
100 x Oil | CFI Plan Apochromat lambda | 1,45 | 0,13 | Fluorescence |
