SLN - Super resolution Light Microscopy and nanoscopy facility

Leica TCS SP8 STED 3X Microscope objectives:  10x, 63x and 100x Excitation wavelengths:  white light laser: 470-670 nm  Semiconductor: 405 nm  8 simultaneous excitation laser lines STED lasers:  592nm, 660nm, 775nm Detectors:  2 Backwards HyD’s  1 Forward PMT  2 Backwards PMT’s Scanning mirrors:  Galvanometric  Resonant Leica TCS SP5 STED CW Microscope objectives:  10x, 20x, 40x, 63x and 100x Excitation wavelengths:  Argon laser: 458, 476, 488, 496, 514 nm  HeNe laser: 543, 633 nm  Ti:Sapphire laser: 750-850nm, pulses duration: 120fs, repetition rate 76MHz STED lasers:  592nm, CW Detectors:  2 Backwards HyD’s  1 Forward PMT  3 Backwards PMT’s  2 Backwards APD’s Scanning mirrors:  Galvanometric  Resonant Abberior Infinity Microscope objectives  60x, WD: 0.15 mm, NA=1.42  10x, NA 0.4 air  40x, WD: 0,18 mm, NA=0.95, variable cover glass correction ring  60x silicone oil, WD: 0.3 mm, NA=1.3, variable cover glass correction ring  40X, WD: 0.15 mm, NA=1.3 oil  Universal Plan Extended Apochromat oil immersion objective with 40x magnification  60x, WD: 0.28 mm, NA = 1.2 water Excitation wavelengths:  Laser Module 405 nm (cw)  Laser Module 485 nm (pulsed, picosecond)  Laser Module 518 nm, (Pulsed, picosecond)  Laser Module 561 nm (pulsed, picosecond)  Laser Module 640 nm (pulsed) STED lasers:  775nm (high-power) and at 595nm Detectors:  Spectral detection modules detector   2 single-photon-counting APD modules   2 single-photon-counting optimized for FLIM detection   APD Array detector Scanning mirrors:  QUAD Beam Scanner: 4 galvo scanners and 4 galvo servo controllers Nikon Confocal C1-Si Microscope objectives:  for confocal/multiphoton: 2x, 10x, 20x, 40x, 60x and 100x  specialized for multiphoton (water inmmersion): Olympus 25x , 1.05 NA and Nikon 25x, 1.10 NA Excitation wavelengths:  Argon laser: 457, 477, 488, 514nm  Diode lasers 405, 561, 635nm  Ti:sapphire 750-850nm  Femtosecond pulsed fiber laser 1550nm  OPO: 1650nm-1750nm Detectors:  1 Forward multiphoton PMT, Large area IR Photodiode, Vis. photodiode  2 Backwards multiphoton PMT’s  2 Backwards confocal PMT’s and 32 spectral detectors Scanning mirrors:  Galvanometric Renishaw InVia Raman micro-spectrometer Microscope objectives:  5x, 20x long working distance, 20x, 50x and 100x Excitation wavelengths:  785nm 300mW  532nm 100mW Light Sheet Fluorescence Microscopes (4 different systems) Microscope objectives:  for excitation: 10X, 0.3 NA and 10X, 0.45 NA  for collection (water inmmersion): 20X, 0.5 NA and 25X, 1.05 NA Excitation wavelengths:  Diode lasers 405nm, 488nm, 638nm  DPSS laser 532nm, 651nm  Ti:Sapphire 750-850nm  Femtosecond fiber laser 1030nm Detectors:  Hamamatsu Orca R2  Hamamatsu Orca Flash 4 Nikon N-STORM microscope Microscope objectives:  10x, 40x, 60x and 100x Excitation wavelengths:  405, 488, 561, 647 nm Detectors  Camera: EMCCD IXON 897

The SLN Facility at ICFO provides: Stimulated emission depletion microscopy: Our STED systems allows 3D super-resolution capabilities in up to 3 optical bands (up to 3 depletion lasers). The single molecule detector (SMD HyD) provides a high detection efficiency. Resonant scanners provide fast scanning capabilities up to videorate at 512×512 pixels. The system includes a white light laser source as well as a 405 nm semiconductor laser. Techniques: STED 3D and 3 colors, Confocal, FLIM, Full-hyper spectral (6D) laser scanning confocal, tau-STED, RESCUE, DYMIN, MINFIELD, easy3D Multimodal microscopy: Our multimodal workstation is built on top of a commercial Nikon Confocal C1-Si. This system contains an additional scanning head and a double filter turret from where a MIRA 900F Ti:Sapphire fs laser is coupled. This allows the simultaneous implementation of multiphoton imaging techniques such as TPEF, SHG/PSHG, THG, as well as the possibility for performing laser surgery or optical tweezers.Techniques: Confocal, hyperspectral confocal, TPEF, 3PEF, SHG, PSHG, THG, Femtosecod nanosurgery,optical tweezers Raman micro-spectrometer: Our Raman microscope includes two excitation lasers: 785nm and 514nm. The system is fully automatized and allows switching the excitation light and adjust the laser power by using the software Wire. The system includes the capability of auto-alignment and line streaming.Techniques: Confocal Raman spectroscopy, Surface Enhanced Raman Spectroscopy Light sheet microscopy: Our LSFM systems are custom-made microscopes designed to explore large microscopic 3D samples like organoids or small embryos. They are highly versatile and can be used for adressing diverse biological questions thanks to its modular fashion. They can be set to work with Gaussian and Bessel beams in the linear and non-linear regime. Volumetric images up to 50 vols/s can be achieved.Techniques: TPEF, Digital Scanned Light Sheet Microscopy, Confocal Line Scan, fast volumetric imaging, optical clearing, high throughput, ultramicroscopy. Stochastic optical reconstruction microscopy: Our STORM system is based in a TIRF configuration with 4 colors laser bench. The focusing position stability is given by the Perfect Focus System and it is fully motorized. Multicolor Single Molecule Localization acquisitions can be obtained by using the high sensitivity of the EMCCD and 3D Localisation is obtained by introducing the cylindrical lens.Techniques: STORM, PALM, SPT, dSTORM.

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