UMIF

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

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Visitron SD-TIRF

Visitron SD-TIRF

 

Location: Campus Forschung (N27), 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): 1.100


20x CFI Plan Fluor DLL Phase

NA: 0.51

WD (mm): 2.1

Pixel size in image (µm): 0.550


40x CFI Plan Fluor Oil

NA: 1.30

WD (mm): 0.2

Pixel size in image (µm): 0.275


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): SpinningDisk: 0.183; SORA disk: 0.065


100x CFI Plan Apo Lambda

NA: 1.45

WD (mm): 0.13

Pixel size in image (µm): spinningDisk: 0.110; SORA disk: 0.039

Cameras

2x Photometrics Prime 95B (back-illuminated sCMOS, 11µm pixel-size, 1200x1200 pixels)

Laser lines (nm)

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

Spinning Disk unit

Yokogawa CSU W-1 SoRa in dual-camera configuration

1) Confocal disk (50µm pinholes)

2) SoRa disk for super-resolution

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-VIS illumination 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 BioPrecision3 LM

Auto-focus Nikon PerfectFocus system
Actively damped optical table Newport

Applications

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