ICCE Equipment - Nikon Ti2 TIRF DualCam

Widefield SystemsNikon Ti2 TIRF DualCam

Room: S05 R01 H24

Main Applications

  • inverted widefield fluorescence microscope
  • 4-color TIRF
  • FRAP
  • simultaneous dual color imaging
  • fast multi-dimensional live cell + fixed specimen imaging
  • multi-position acquisition

 

Funding

Nikon TIRF DualCam microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 361032973.

 

Technical Specifications

Microscope and Accessories

  • inverted Nikon Ti2-E
  • Perfect Focus System PFS4 (hardware autofocus system)
  • motorized xy-stage
  • Nikon Ti LAPP system with modules:
    • epi-fluorescence
    • H-TIRF for fully automated TIRF adjustments
    • FRAP for photobleaching and photoactivation/conversion experiments
  • okolab bold line cage incubator with control device (temperature & CO2 control for live-cell imaging)

Cameras and TuCam

  • 2 x: Andor Technology iXon Life DU-888 back-illuminated EMCCD camera, 1024 x1024 pixels imaging array, 13 x 13 µm pixel size, 13.3 x 13.3 mm sensor format, 16-bit digitization, 56 fps at full frame
  • Andor TuCam Dual Camera Imaging adapter, (GFP/mCherry: Brightline HC 525/50, Brightline HC 609/54, Beamsplitter R561)

Light sources

  • Lumencor Sola SE II solid state white light illumination
  • LED transmitted light module

Lasers

Omicron Light Hub-4 compact laser beam combiner with AOTF laser modulator and fast fiber switch with two fiber outputs (TIRF & FRAP)

  • Diode Laser cw LuxX 405 nm (120 mW)
  • Diode Laser cw LuxX 488 nm (250 mW)
  • DPSS Laser cw Jive 561 nm (300 mW)
  • Diode Laser cw LuxX 638 nm (200 mW)

Fluorescence filters & dichroics

Filter cubes for LED epifluorescence

name Fluorochromes excitation [nm] dichroic [nm] emission [nm]
HC LED 400 DAPI/Alexa405/EBFP 406/15 425 450/28
HC LED 465 EGFP/Alexa488 460/60 495 525.5/23.5
HC LED 565-585 RFP/mCherry/Alexa594 562/40 593 624/40
LED 615-635 ET Cy5/Alexa647 620/60 660      700/75

Quadband dichroic and emission filters for TIRF

name Fluorochromes excitation [nm] dichroic [nm] emission [nm]
NSTORM QUAD multiple laser lines

420-481

497-553

575-628

667-792

446/32.5

523.5/42

603/44

730/112
TIRF emission filter #1 EGFP/Alexa488     525/50
TIRF emission filter #2 RFP/mCherry/Alexa594     600/50
TIRF emission filter #3 Cy5/Alexa647     705/72

Objectives

name

magnification

 NA immersion
CFI Plan Apo λ 10x/0.45 10x 0.45 dry
CFI Plan Apo λ 20x/0.75 20x 0.75 dry
CFI Plan Apo λ 40x/0.95 CC 0,25-0,16 40x 0.95 dry
CFI Apochromat TIRF 60x/1.49 60x 1.49 oil
CFI Apochromat TIRF 100x/1.49 100x 1.49 oil

 

Software

Nikon Imaging software NIS Elements AR (Advanced Research)