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Molecular Imaging Center (MIC)
Widefield system with SMLM

Nikon Eclipse Ti2

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This instrument is part of the NORBRAIN nanoscopy facility and is organized under MIC. 

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. 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

 

(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

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

1,20

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