TIRF 1 Nikon-Roper ILAS

Joanna Lipka is studying AMPA-receptor in hippocampal neurons using FRAP assay

Primary tasks:
TIRF microscopy, Single Molecule Imaging, FRAP, Photo-Activation, Multi-position acquisition,
Electrical Stimulation, Perfusion Experiments, Phase Contrast Time Lapse Microscopy

Microscope: Nikon Eclipse TE2000 with Perfect Focus System

Objectives:
Apo TIRF 100x/1.49 oil, Plan Apo VC 100x/1.4 oil, Plan Fluor 40x/1.30 oil, Plan Apo 40x/1.0 oil,
Plan Apo 20x/0.75 dry

Beamsplitter: DV 2 (two-channel, simultaneous-imaging system)

Camera:
Photometrics Evolve 512 EMCCD
Photometrics CoolSNAP HQ2 CCD

Lasers:
Vortran 405 nm (100 mW)
Cobolt Calypso 491 nm (100 mW)
Cobolt Jive 561 nm (100 mW)

FRAP: ILas0 system (Roper Scientific France/Institut Curie)

Filters:
ET-DAPI (49000), ET-CFP (49001), ET-GFP (49002), ET-YFP (49003), ET-DSRed (49005), ETCy5
(49006), ET-mCherry (49008), ET-CFP/YFP (59017), ET-GFPmCherry (59022) – all Chroma

Stage: Prior

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

Heating/CO2: Tokai Hit INUG2E-ZILCS Stage Top Incubator

Software: MetaMorph 7.7

Other equipment:
Narishige NT-88-V3 micromanipulator, Master-8 (A.M.P.I.), High-current stimulus isolator (World
Precision Instruments), Pressurized flow system with computer-controlled valves (Harvard
Apparatus)

Location: Z-515
Contact Person: Ilya Grigoriev