Nikon iLAS2 TIRF/FRAP

Location: I23.U1.015 (Sunstone building)

System description and specifications

Nikon Ti2E stand equipped with iLas2 (GATACA) 360° / Ring / Azimuthal TIRF module. This module is using a pair of fast galvanometers to be able to spin the laser in the back focal plane of the objective several times during a single exposure time. That way artifacts such as fringes or rings caused by laser interference patterns are averaged out.

  • 360° TIRF imaging
  • FRAP (with all laser lines)
  • Photoactivation / -conversion (with all laser lines)
  • Micro-patterning
  • Widefield imaging
  • laser angle sweep

Microscope and sample conditions

  • Temperature controlled chamber
  • CO2 supply

 

 

Light sources

  • CoolLED pE300 White MB for widefield imaging and visual inspection of the sample
  • Halogen lamp for visual inspection of the sample
  • Omicron LightHUB Ultra laser system TIRF imaging and FRAP: 405nm, 488nm, 561nm, 640nm

Cameras

  • 2 x Andor iXon Life 888 Back Illuminated EMCCD camera 1024 x 1024 active pixels, pixel size 13 μm x 13 μm, sensor area: 13.3 mm x 13.3 mm; 18.8 mm diagonal
  • Front illuminated CMOS monochromatic camera for back focal plane imaging, 2048 x 1536 pixels, pixel size 3.45 µm

Objectives

  • 20X / NA 0.75 / WD 1.00 mm Lambda CFI P –Apo (MRD00205)
  • 60x/ NA 1.49/ WD 0.13  TIRF oil CFI Apochromat, with correction collar for coverglass thickness (0.13-0.20 mm), chromatic correction 435-1064nm (MRD01691)
  • 100x/ NA1.49/ WD 0.12 oil TIRF oil CFI Apochromat, with correction collar for coverglass thickness (0.13–0.20 mm), chromatic correction 435–1064nm (MRD01991)

Dual-Cam filter option

Cairn TwinCam camera splitter

  1. TwinCam Cube dichroic mirror red/farred 643 nm
  2. TwinCam Cube dichroic mirror green/red,farred: 560 nm
  3. TwinCam Cube dichroic mirror blue/green: 488 nm

Bandpassfilters for TwinCam

450/50 ET Bandpass

525/50 ET Bandpass

595/50 H Bandpass

635 Longpass-Filter (Transmission 660 – 1200 nm)

700/75 H Bandpass

Filtercubes for Epifluorescence

  1. LED-FITC-A Filter Cube EX 474/27, EM 525/45
  2. LED-TRITC-A Filter Cube EX 554/23, EM 609/54
  3. LED-Cy5-A Filter Cube EX 628/40, BA 692/40

Software

NIS elements Ar 5.42.04

Computer: HP Z4 workstation

  • Intel Xeon 3.9GHz 8Cores
  • RAM:128GB DDR4
  • 512GB OS Drive
  • 2x 2TB Fast Storage
  • 8TB Data Storage
  • NVIDIA Quadro RTX5000 (16GB)

System manual and additional material is available on SeaFile (login required).

contacts

  1. Doreen Milius
  2. Yann Cesbron
  3. Nasser Darwish Miranda