Nikon iLAS2-02 TIRF/FRAP

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.

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Description

Location: I04.U1.019 (Bertalanffy Foundation Building)

Coming Soon…

System description and specifications

  • 360° TIRF imaging
  • FRAP (with all laser lines)
  • Photo-activation / conversion (with all laser lines)
  • Micro-patterning
  • Widefield imaging
  • Motorized stage
  • Perfect focus system
  • Laser angle sweep

Microscope and sample conditions

  • Temperature controlled chamber
  • CO2 supply

Light sources

  • CoolLED pE-300 white MB LEDs for widefield fluorescence imaging and visual inspection of samples
  • LED light source for transmitted light visual inspection of samples (+ GIF filter)
  • Omicron LightHUB Ultra laser system for TIRF imaging & FRAP: 405 nm (120 mW), 488 nm (100 mW), 561 nm (100 mW), 640 nm (100 mW)

Cameras

  • 2 x Photometrics Kinetix 22 mm sCMOS camera 2400 x 2400 active pixels, pixel size 6.5 μm x 6.5 μm, sensor area: 15.6 mm x 15.6 mm; 22.1 mm diagonal
  • Digital Sight 100 front illuminated CMOS colour camera for back focal plane imaging, 4864 x 3648 pixels, pixel size 2.4 µm

Objectives

  • 20x / NA 0.8 / WD 0.80 mm CFI Plan Apochromat lambda D (MRD70270)
  • 60x / NA 1.49 / WD 0.13 mm oil CFI Apochromat TIRF, with correction collar for coverglass thickness (0.13-0.20 mm), chromatic correction 435-1064 nm (MRD01691)
  • 100x / NA 1.49 / WD 0.12 mm oil CFI Apochromat TIRF, with correction collar for coverglass thickness (0.130.20 mm), chromatic correction 4351064 nm (MRD01991)

Dual-Cam filter option

CAIRN TwinCam camera splitter (0.76x magnification)

  1. TwinCam cube dichroic mirror red/far red 643 nm
  2. TwinCam cube dichroic mirror green/red,far red: 560 nm
  3. TwinCam cube dichroic mirror blue/green: 488 nm (on request)

Bandpassfilters for TwinCam

  • 450/50 ET bandpass
  • 525/50 ET bandpass
  • 595/50 ET bandpass
  • 635 longpass filter (transmission 660 – 1200 nm)
  • 700/75 ET bandpass

Filtercubes for Epifluorescence

  1. LED-DAPI-B filter cube Ex 392/23, Em 447/60
  2. LED-FITC-A filter cube Ex 474/27, Em 525/45
  3. LED-TRITC-A filter cube Ex 554/23, Em 609/54
  4. LED-Cy5-A filter cube Ex 635/18, Em 680/42

Software

NIS-Elements AR 6.20.03

Computer: HP Z4 G5 workstation

  • Intel Xeon W3 4.3 GHz 8 Cores
  • RAM:128 GB DDR5
  • 512 GB OS Drive
  • 2x 2 TB SSD Fast Storage
  • 8 TB Data Storage
  • NVIDIA Quadro RTX4000 (20GB)

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

Additional information

Building

i06

Camera / Detector

sCMOS

Fluorescent Lightsource

Laser: 405±5 nm, Laser: 488±5 nm, Laser: 561±5 nm, Laser: 640±5 nm, LED: 355-365nm, LED: 430-450nm, LED: 545-555nm, LED: 640-650nm

System_Specification

Illumination: Fluorescence, Illumination: DIC, Incubation: heating chamber, Incubation: CO2/N2-gas mixer (hypoxia), Automated Stage, Z-piezo / galvo Stage, Focus Stabilization, Dual Camera (see detectors)

Technology / Application

360° / Ring / Azimuthal TIRF, FRAP, In-vivo imaging, Micro-patterning, Photoactivation / -conversion, SAIM, SR: SMLM (20nm)