Visitron iLAS2 TIRF/FRAP

Posted onAuthorGabriel Krens

Visitron iLAS2 TIRF/FRAP

Automated epifluorescence microscope, equipped with iLas 2 360° / Ring / Azimuthal TIRF module and (GATACA): 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 ring caused by laser interference patterns are averaged out.

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Description

Location: room I04.U1.018 (BFB)

  • 360° TIRF imaging
  • FRAP (with all laser lines)
  • Photoactivation / -conversion (with all laser lines)
  • Micro-patterning
  • Widefield imaging
  • DIC imaging (in combination with epifluorescence)

Microscope & sample conditions

  • Temperature controlled chamber
  • CO2 supply

Light sources

  • Laser lines: 405 nm, 488 nm (100 mW), 561 nm (100 mW), 640 nm (150 mW) – Laser Quad Band Filter for TIRF application: 405/488/561/640 nm
  • Lumencor Spectra-X Solid State/LED – light source: wavelength range 380-680 nm
  • Halogen lamp

Objectives

  • 10x / 0.3 air (WD 2.1 mm)
  • 20x / 0.4 air (WD 8 mm)
  • 20x / 0.8 air (WD 0.55 mm)
  • 63x / 1.46 oil (WD 0.1 mm) – TIRF objective
  • 100x /1.46 oil (WD 0.1 mm) – TIRF objective

Cameras

2 x Photometrics Evolve 512 EMCCD (512 x 512 pixels, 16 x 16 μm2)

Dual-Cam Filter Options

  • Cairn TwinCam camera splitter (2 dichroic filter cube options) :
  1. cube: LP565
  2. cube: LP635
  • Band-pass filters
  1. 525/50
  2. 595/50
  3. 630/75
  4. 670/50
  5. 690/50

Software

Visiview 4.4

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

Additional information

Building

i04
Fluorescent Lightsource

Laser: 470-670nm WLL, Laser: 405±5 nm, Laser: 488±5 nm, Laser: 640±5 nm
Camera / Detector

EMCCD
System_Specification

Illumination: Fluorescence, Illumination: Bightfield, Illumination: DIC, Incubation: heating chamber, Incubation: CO2-gas mixer, Automated Stage, Focus Stabilization, Dual Camera (see detectors)
Technology / Application

360° / Ring / Azimuthal TIRF, FRAP, FRET, Time-lapse recording, Multi-position, In-vivo imaging, Micro-patterning, Photoactivation / -conversion

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