Leica 4TuneDualIRDIVE STELLARIS 8 FALCON [Grant EQC2021-007527-P funded by MCIN/AEI /10.13039/501100011033 and by European Union NextGenerationEU/PRTR]

Applications

This upright microscope, first in Europe, is equipped with a double-line multiphoton laser and 4 spectral photon counting NDD detectors allowing dynamic selection of emission windows from 410 to 750 with 10 nm steps [Leica 4Tune Dive]. It is combined with a confocal white laser, 405 nm diode and photon counting Hybrid detectors [Leica Falcon] and completed with an incubator chamber for CO2, N2 and O2 control and a variety of sample adaptors and stages. Therefore, this system can be used for multiparametric imaging including 1P, 2P, SHG, THG and 1P- and 2P-FLIM on cells, tissues, cleared organs and intravital models in 3D, to detect and characterize simultaneously in the same sample complexes structures and endogenous components (fibers, collagen, lipids) and multiple fluorescent and auto-fluorescent biomolecules. 

NOTICE: Due to the variety of applications, please contact the Unit staff with enough advance for an appropriate set-up of the system.

SoftwareLAS X, Lightning (deconvolution), Navigator (tiles), mathematical FLIM fitting up to 5 components and PhasorFLIM modules are also included.
Objectives
  • Revolving nosepiece for 25mm diameter objectives
    • HC PL APO 10x/0.40 CS2. Dry (Under request)                             
    • HC PL APO 20x/0.75 Multi-immersion CS2           
    • HC FLUOTAR L 25x/0.95 Water immersion (dipping) VISIR
    • HC APO L 40x/0.80 Water immersion (dipping) UVI         
    • HC PL APO 40x/1.30 Oil immersion
    • HC PL APO L63x/0.90 Water immersion (dipping) UVI CS2         
    • PL APO 63x/1,40 Oil immersion CS2         
  • Lens holder for 32mm diameter objectives
    • HC APO L 20x/1.00 Water immersion (dipping)
Incubator

With a chamber enclosing the microscope, digital control of temperature, CO2, N2 and O2 allowing long term in-vivo experiments.
CO2 range from 0 to 20% and humidity control.
For the use of gases such as O2, N2, carbogen... Consult the unit staff.

Stages
  • Stage adapted for large-sized live and fixed samples motorized XY and fixed Z to keep the sample stable.
  • Motorized Z focus using wide-range objective nosepiece focus at least up to 13mm.
    • Manual control of the height (Z), range of 50mm.
    • Fully integrated into the Navigator software, allowing execution of multi-position and regular and irregular shaped mosaics
  • Support for in-vitro applications with threaded holes to hold the samples.
  • Galvanometric stage for fast and precise movement in Z. Movement range of 1500μm; adjustable in <0.5nm steps; reproducibility <40nm.

 

Sample Setup

Multiwell holders (size P6, P12, etc. ....) P35 plates and adapter for slide size sample holders.
Notice that some holders are only available for some objectives (consult the Unit staff).

Illumination Source
  • Pulsed white light laser (440 - 790 nm)
  • 405nm UV laser
  • Pulsed infrared laser with two independent excitation lines
    • tunable line (tunable excitation range between 680 - 1300nm)
    • fixed line (Excitation range 1045nm)
  • LED-based fluorescence light source (390 - 680nm)
Detectors/Cameras
  • CONFOCAL DETECTION SYSTEM 
    • Three hybrid photon counting  detectors (HyD S). Detection range 410 - 850nm 
    • Two hybrid photon counting  detectors (HyD X, recommended for FLIM). Detection range 410 - 750nm 
  • MULTIPHOTON DETECTION SYSTEM 
    • 4 NDD spectral photon counting  detectors (recommended for FLIM). Detection range 410 - 750nm
  • TRANSMITTED LIGHT DETECTION SYSTEM 
    • 2 TPMTs
ConfigurationUpright.
FiltersTriple or individual filters (UV, green, red) on request.
External Accesshttps://www.cnic.es/en/investigacion/2/1187/interes