TIRF Nikon Ti – Roper iLas FRAP/Photoablation system

Andrea Marques and Stine Morthorst are studying the exocytosis of Rab6 vesicles

Primary tasks:
TIRF microscopy, Single Molecule Imaging, FRAP, Photo-Activation, Photo-Ablation (532 nm)

Microscope: Nikon Eclipse Ti with Perfect Focus System

Objective: Apo TIRF 100x / 1.49, oil, WD=0.09-0.16 (Nikon, Japan)

Beamsplitter: Optosplit III (allows two or three color simultaneous imaging)

Camera:

Evolve 512 EMCCD (512×512 pixels) (Teledyne Photometrics, USA)
Prime BSI sCMOS camera (2048×2048 pixels) (Teledyne Photometrics, USA)

Lasers:

Vortran Stradus 405 nm (100 mW) (Vortran, USA)
Vortran Stradus 488 nm (150 mW) (Vortran, USA)
Cobolt Jive 561 nm (100 mW) (Cobolt, Sweden)
Vortran Stradus 642 nm (110 mW) (Vortran, USA)
Teem Photonics 532 nm Q-switched pulsed laser (for photoablation) (Teem Photonics, France)

FRAP: ILas system (Roper Scientific France/ PICT-IBiSA, Institut Curie; now Gataca Systems, France)

Filters:

ET-DAPI (49000), ET-CFP (49001), ET-GFP (49002), ET-YFP (49003), ET-DSRed (49005), ET-Cy5 (49006), ET-mCherry (49008), ET-CFP/YFP (59017), ET-GFPmCherry (59022) – all Chroma

Stage: ASI motorized stage MS-2000-XY System for Inverted Microscope Nikon Te/Ti 2000

Filter Wheels: Sutter LB10-3 (excitation filter wheel with smart shutter; emission filter wheel; smart shutter for transmission light)

Heating: Tokai Hit STXG-PLAMX-SETZ21L Stage Top Incubator

Software: MetaMorph 7.10

Resolution: 82 nm in 1 pixel (Evolve camera) / 64 nm in 1 pixel (Prime BSI camera)

Installation: 2011

Location: Z-526
Contact Person: Ilya Grigoriev