UMIF

UKE Microscopy Imaging Facility (DFG Research Infrastructure Portal: RI_00489)

Visitron SD-TIRF

Visitron SD-TIRF

 

Location: room 02.053, second floor

Laser lines: 405nm/445nm/488nm/515nm/561nm/640nm

Software: VisiView

Live cell Imaging: Add-on environmental chamber with temperature, humidity and CO2 control

 

 

Microscope Nikon Eclipse TiE
Objectives

10x CFI Plan Fluor DL Phase

NA: 0.30

WD (mm): 15.2

Pixel size in image (µm): 0.800 (w/ 2x intermediate magnification)


20x CFI Plan Fluor DLL Phase

NA: 0.51

WD (mm): 2.1

Pixel size in image (µm): 0.400 (w/ 2x intermediate magnification)


40x CFI Plan Fluor Oil

NA: 1.30

WD (mm): 0.2

Pixel size in image (µm): 0.200 (w/ 2x intermediate magnification)


60x Apo TIRF (corr.) Oil

NA: 1.49

WD (mm): 0.13 (CS: 0.10-0.22 @ 23 or 37 °C)

Pixel size in image (µm): 0.133 (w/ 2x intermediate magnification)


100x CFI Plan Apo Lambda

NA: 1.45

WD (mm): 0.13

Pixel size in image (µm): 0.080 (w/ 2x intermediate magnification)

Cameras

2x evolve-EM 512 (back-illuminated EM-CCD, 16µm pixel-size)

Gain: 7 e-/ADU

Laser lines (nm)

Solid-state: 405 / 445 / 488 / 515 / 561 / 640

Spinning Disk unit Yokogawa CSU W-1 in dual-camera configuration
Emission filters spinning disk (for laser-related fluorescence)

Camera 1:

  • DAPI, ET460 (W50)
  • CFP ET470 (W24)
  • GFP, ET525 (W50)

Camera 2:

  • YFP ET535 (W30)
  • mCherry, ET609 (W54)
  • CY5 ET700 (W75)
Dichroic in spinning disk unit

405/488/561/640 (used with the respective channels)
445/515/640 (used for 445, 515 and 640 channels)

Dichroic for dual camera mode

561LP
514LP

Fluorescence filters in microscope stand

Filtersystem (em.-color, dye): excitation | beamsplitter | emission

  • DAPI (blue; DAPI):
    BP 325-375 | DM 400 | BP 435-485

  • GFP (green; AF488, GFP):
    BP 450-490 | DM 495 | BP 500-550

  • TxRed (red; AF568, mCherry):
    BP 540-580 | DM 585 | BP 593-668

Dichroic mirrors in microscope stand for TIRF and FRAP

TIRF-FRAP1:

  • 405/488/561/640 rpc BS (Chroma; 2mm)

TIRF-FRAP2:

  • 442/514 rpc BS (Chroma; 2mm)
UV-lamp SOLA-SM Light Engines, white light LED, 380-680nm
Transmitted light lamp precisExcite, High-Power 525nm LED
Software VisiView
Incubation unit okolab bold line
FRAP-TIRF unit Roper iLAS2 for FRAP&TIRF with all laser lines
Piezo focus drive Ludl NanoPrecision PiezoZ, 350µm travel range
Motorized XY stage

Ludl BioPrecision2

Auto-focus Nikon PerfectFocus system
Actively damped optical table  Newport

Applications

SpinningDisk, TIRF, Live imaging, FRET, Widefield, FRAP, Photoactivation/-conversion
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