Room: S05 T04 B97

Main Applications

• confocal (point) laser scanning microscopy, optical sectioning
• FLIM
• F(C)CS, FLCS
• laser microirradiation (e.g. DNA damage) via UVA laser ablation system (355 nm)
• Photoactivation / Photoconversion
• Confocal super-resolution with Leica Lightning (Detection and Image Information Extraction/ Deconvolution for “confocal super-resolution” down to 120 nm)
• FRAP
• FRET (FLIM-FRET, acceptor photobleaching & sensitized emission)
• λ² Excitation-Emission scans
• multi-dimensional live cell + fixed specimen imaging
• multi-position, tile scan / Mosaic
• automated imaging & screening (Leica HCS A, Leica Navigator)

Funding

Leica TCS SP8X FALCON microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 397277702.

Technical Specifications

Microscope and Accessories

• inverted Leica DMi8 CS
• Leica Adaptive Focus Control (AFC)
• motorized xy-stage
• SuperZ Galvo
• SuperZ stage accessories: Microtiterplate insert, adapter for petri dishes (36 mm dishes) and universal sample holder
• external fluorescence lamp LQ-HXP 120 LEJ
• LED lamp for transmitted light and equipment for confocal DIC
• controlled environmental conditions for live cell imaging (temperature, CO₂)

Lasers

• continuous wave lasers
  • Diode Laser (50 mW): 405 nm DMOD
  • Argon Laser (65 mW): 458 nm, 476 nm, 488 nm, 496 nm, 514 nm
• pulsed lasers
  • WLL E laser + pulsepicker: 470 nm to 670 nm, avg. power 1 mW, integrated pulsepicker: 80 MHz, 40 MHz, 20 MHz, 10 MHz
• Laser ablation system (UVA laser)
  • UGA-42 Caliburn 355/42 (Rapp OptoElectronic), 355 nm

Scanners

• non-resonant (conventional/variable) Scanner: 1 – 1800 Hz (7 fps @ 512 x 512 px, 84 fps @ 512 x 16 px), line frequency up to 3600 lines/second (bidirectional), max. scan format/image resolution 8192 x 8192 px (64 Mpixel), scanfield 22 mm (SFZ)
• resonant Scanner (ultra-fast, reduced photodamage): 8000 Hz (28 fps @ 512 x 512 px, 290 fps @ 512 x 16 px), line frequency up to 16000 lines/second (bidirectional), max. scan format/image resolution 1248 x 1248 px (> 1 Mpixel), scanfield 13 mm (SFZ)

Detector System

• 5-channel SP-Detector: 2 HyDs SMD, 1 HyD, 2 PMTs
• 1 PMT for transmitted light (PMT trans)

Beam splitting between excitation and emission light is controlled via AOBS.

Objectives

Filter sets for LQ HXP-120

• Filterset DAPI (Excitation BP 350/50, Dichroic 400, Emission BP 460/50)
• Filterset FITC (Excitation BP 480/40, Dichroic 505, Emission BP 527/30)
• Filterset RHOD (Excitation BP 546/10, Dichroic 560, Emission BP 585/40)

Software

Main CUDA workstation equipped with software modules:

• LAS X Core
• LAS X FCS (acquisition and analysis of FCS, F(C)CS, and FLCS data)
• LAS X Lightning Expert (detection and Image Information Extraction/ Deconvolution for “confocal super-resolution” down to 120 nm)
• LAS X Navigator (fast overviews, automated image acquisition)
• HCS-A Matrix Screener (for automated microscopy, high content screening)
• LAS X Live Data Mode (Interactive data recording allowing job-sequencing and online evaluation)
• LAS X MicroLab (FRET, FRAP, FLIP)
  • FRAP wizard, FRAP XT wizard
  • FRET wizard (acceptor photobleaching & sensitized emission)
• LAS X 3D Visualization
• LAS X Dye Finder (Multi-color restoration, channel unmixing)
• LAS X Environmental control (full control of your experimental conditions, log of the environmental data and monitoring during experiment)
• Rapp OptoElectronic SysCon2 (control and setup of UVA laser ablation system and connection to LAS X via LocalCam plugin)

Offline workstation equipped with software modules:

• LAS X Basic License
• LAS X FLIM offline analysis
• LAS X FCS offline analysis
• LAS X 3D Visualization