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Leica Stellaris 8 DIVE FALCON (Multiphoton/FLIM):

The system enables fluorescence excitation with a dual line laser in the infrared range, multiplexing using spectral information and video-rate FLIM. Optional beam routing to achieve a balance between resolution and penetration depth. The InSight X3 Laser covers a broad excitation range from 680 nm to 1300 nm for optimal penetration, imaging of far red probes, three-photon excitation, and third harmonic generation imaging. The single beam can be combined with the fixed 1045 nm dual beam option. This supports the diverse needs of multimodal imaging. The two synchronized output beams enable easy simultaneous imaging of various fluorescence proteins, genetically encoded calcium indicators, second and third harmonic generation. In 12 kHz resonant scanning mode also fast physiological process can be imaged. STELLARIS 8 DIVE enables the flexible spectral imaging of 4 simultaneous channels. The system also offers a sequential mode, so more than 4 signals can be captured sequentially. The FALCON (FAst Lifetime CONtrast) offers functional imaging and is the first truly integrated solution for Fluorescence Lifetime Imaging (FLIM) and is at least 10x faster than previous solutions. It is fast enough to follow highly dynamic cellular events with lifetime-based biosensors and detect molecular interactions with FRET. The navigator tool provides orientation within the specimen, allows fast overviews, and enables detailed multi-position experiments.

Applications:

Laser:  Spectra Physics Insight X3 Dual

Microscope stand:

Leica DM6 CFS (motorized upright microscope)

Detection:

Confocal detectors: 2 HyD SP gated, 1 HyD S SP gated) (FLIM capable) (380-800 nm)

4TUNE DIVE (4-channel): 4X non-descanned 4Tune HyD RLD detectors (FLIM capable) with free tunable detection range (380-800 nm)

2 transmission detectors: PMT with filter block

FLIM:

SP8 FALCON (FAst Lifetime CONtrast) with LAS X Phasor FLIM Online

Multi-beam mode:

Broad tuning range (680–1300 nm) for maximum flexibility.

Dual output at 1045 nm with optional pre-compensation for simultaneous two color excitation. Dual laser beam routing with VBE.

Laser Scanning:

TCS SP8 Scan head (resonant/non-resonant)

Microscope Stage:

Motorized Scientifica MMBP X/Y stage, Okolab laser security box (can be heated).

Microscope Table:

Anti-vibration table (active dampening) Newport

Computer:

Expert CUDA Workstation, RAID High Power HPZ6G4 Workstation

Software:

LAS X, Navigator, AviaMotion

Others:

EL 6000 (LQ-HXP 120 LEJ) fluorescent light source, three fluorescent filter cubes (DAPI/GFP/RHOD).

Trigger box for the transformation of external signals (ECG and respiration) into trigger pulses for in vivo imaging