Main Applications:

TIRF, 4-color SMT, FPALM, STORM

Person in Charge:

Rainer Kurre

Location:

Building 38, Room E15b, S2-laboratory
Bild "Light Microscopy:BP_TIRF_4_Line.jpg"

Olympus TIRF 4-Line STORM

Filter

Microscope filter cassette
  • Pos. 1: Pentacolor TIRF (blue, green, yellow/orange, red, dark red)
    • Beamsplitter: TIRF pentaband polychroic mirror, 405 nm/488 nm/561 nm/640 nm/730 nm (zt405/488/561/640/730rpc, Semrock)
    • Emitter: pentabandpass, 440 nm/521 nm/607 nm/694 nm/809 nm (BrightLine HC 440/521/607/694/809)
  • Pos. 2: Cy3 + Cy5
    • Beamsplitter: polychroic mirror, 532 nm/642 nm (zt532/642rpc)
    • Emitter: longpass at 532 nm
  • Pos. 3: 3-Line TIRF plus FRAP
    • Beamsplitter: polychroic mirror, 488 nm/561 nm/633 nm (zt488/561633,405tpc)
  • Pos. 4: 532 TIRF (NileRed)
    • Beamsplitter: dichroic mirror, 532 nm (zt532rdc flat)
    • Emitter: longpass at 532 nm
  • Pos. 8: IX2-MDICT (analyzer for DIC imaging)

Filter cubes for FRAP module
  • Cube 1: 20/80 FRAP/TIRF
    • Beamsplitter: dichroic mirror, 20%/80% (IX3-FRAP20-80BS)
  • Cube 2: FRAP with 405 nm
    • Beamsplitter: dichroic mirror, 405 nm (H 405 LPXR superflat)
    • Emitter: QuadLine Rejectionband (zet405/488/561/640m)
  • Cube 3: Photonmanipulation with 445 nm
    • Beamsplitter: dichroic mirror, 442 nm (HC BS R442 1 lambda PV flat)
    • Emitter: longpass filter (458 LP Edge Basic)


Fast Emission Filter Wheel (Sutter Lambda 10-3)
  • Pos. 1: blue channel (DAPI, BFP): bandpass 423 - 468 nm (BrightLine HC 445/45)
  • Pos. 2: green channel (GFP): bandpass 502 - 538 nm (BrightLine HC 525/35)
  • Pos. 3: orange channel (TMR, mCherry): bandpass 575 - 625 nm (600/50 ET)
  • Pos. 4: red channel (Cy5, Atto 655): bandpass 660 - 710 nm (685/50 ET)
  • Pos. 5: far red channel (Cy7, DY-752): bandpass 768 - 849 nm (BrightLine HC 809/81)
  • Pos. 6: bypass for image splitter
  • Pos. 7: dual Bandpass (GFP & mCherry): bandpass 500-550 nm, 615-700 nm(HC 527/645)
  • Pos. 8: notch filter at 647 nm for Cy3 + Cy5 filter cube (zet647NF)
  • Pos. 9-10: empty position

MultiSplit filter cubes
Cube 1a:
  • Dichroic beamsplitter at 600 nm (NileRed) (H 600 LPXR superflat, 2mm)
Cube 1b:
  • Dichroic beamsplitter at 640 nm (Cy2/Cy3/Cy5/Cy7) (H 643 LPXR superflat, 2mm)

Cube 2:
  • Dichroic beamsplitter at 730 nm (T 725 LPXR, 2mm)
  • Emission filter
    • far red channel (Cy5):
      • bandpass, 660 -710 nm (ET 685/50) in combination with Cube 1a
      • bandpass, 600-700 nm (BrightLine HC 650/100) in combination with Cube 1b
    • NIR channel (Cy7): bandpass, 769 - 850 nm (Brightline HC 809/81)

Cube 3a:
  • Dichroic beamsplitter at 488 nm (Cy2/Cy3) (H 488 LPXR superflat, 2mm)
  • Emission filter
    • Green channel (GFP, Cy2): bandpass,  502 - 538 nm (BrightLine HC 520/35)
    • Orange channel (TMR, mCherry): bandpass 582 - 619 nm, (BrightLine HC 600/37)

Cube 3b:
  • Dichroic beamsplitter at 532 nm (NileRed) (zt532rdc, 2mm)
  • Emission filter
    • Green (GFP): bandpass, 500 - 520 nm (BrightLine HC 510/20)
    • Orange (TMR): bandpass, 502 - 538 nm (575/50 ET)