UMIF

UKE Microscopy Imaging Facility

Olympus TIRFM

Olympus TIRFM

 

Location: room 01.068, first floor

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

Software: Cell Sense

Live cell imaging: Environmental chamber with temperature, humidity and CO2 control

 

 

Microscope Olympus IX-81
Objectives

10x U Plan Semi Apo PH1

NA: 0.30

WD (mm): 10

Pixel size in image (µm): 1.161


20x UPlan FL N

NA: 0.50

WD (mm): 2.1

Pixel size in image (µm): 0.580


40x UPlan S Apo

NA: 0.90

WD (mm): 0.18 (CS: 0.11-0.23 @ 23 or 37 °C)

Pixel size in image (µm): 0.290


60x Plan Apo N TIRF Oil

NA: 1.45

WD (mm): 0.1 (CS: 0.13-0.19 @ 37 °C)

Pixel size in image (µm): 0.194


100x UPlan Apo HR TIRF Oil

NA: 1.50

WD (mm): 0.11 (CS: 0.13-0.19 @ 37 °C)

Pixel size in image (µm): 0.116


150x U Apo N TIRF Oil

NA: 1.45

WD (mm): 0.07 (CS: 0.13-0.19)

Pixel size in image (µm): 0.078

DIC prisms

Condenser:

  • DIC20, DIC40, DIC60

Slider:

  • IX2/U-DICTS

Cameras

Hamamatsu Flash 4.0 V2

Gain: 0.5 e-/ADU

Laser lines (nm)

Solid-state: 405 / 488 / 561 / 640

Fluorescence filters fluo-turret:

Filterset no. (em.-color; dye): excitation | beamsplitter | emission

  • 488 525.40 (green; GFP, AF 488):
    BS 488 | emission: 525/40
  • 561 630.75 (red; RFP, AF568):
    BS 561 | emission: 630/75
  • TIRF (blue/green/red/far-red):
    emission: 420-480/500-550/570-620/LP655
  • GFP/RFP [OBS-U-M2GFPDSRED/HC] (green/red; AF488, AF568)
  • DaFiTri [U M3DAFITR] (blue; DAPI, AF488, AF569)
Fluorescence excitation filters UV-lamp

Filterset name (ex.-color; dye): excitation (nm)

  • DAPI (UV; DAPI, filipin): 350
  • FITC (blue; AF 488): 485
  • TxRed (green; AF568): 585
  • GFP (blue; GFP): 470
  • RFP (green; RFP): 556

UV-Lamp

75W xenon lamp

Halogen lamp

100W 12V, standard housing

Acquisition software

OlympusCell Sense V1.14

TIRF light path 

Motorized, for 405/488/561/640nm excitable fluorophores

Incubation unit

Evotec closed large chamber for temp./CO2/humidity
Hardware autofocus

ZDC2

Actively damped optical table  Newport

Applications

TIRF, Live imaging, Widefield
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