Leica SP8 STED

Brief description:

This confocal Leica TCS SP8 microscope consists of  an inverted stand equipped with a STED 3X unit (STimulated Emission Depletion) and a setup for time resolved fluorescence microscopy including a FLIM (Fluorescnece LIfetime Microscopy) and a  FCS/FCCS unit (Fluorescence /Cross/ Correlation Spectroscopy).

Microscope:

Leica DMI6000B-CS with blue/green/red epifluorescence illumination for visual observation through the eyepiece; Condensor (Air 0.55NA)

Illumination:

• Laser unit for confocal acquisition (AOBS system): 458, 477, 488, 496, 514 nm (Argon laser), 405 nm, 561 nm, 633 nm
• White Light Laser: tunable 470–670 nm
  
• STED Lasers: 592 nm, 660 nm, 775 nm
  
• FLIM Laser: 440 nm

Objectives:

• 10x 0.4NA HC PLAN APO CS
  
• 20x 0.75 NA, Oil/W/Glyc, HC PLAN APO CS2
  
• 63x 1.4NA Oil  HC PL APO CS2
• 63x 1.2NA W HC PL APO CS2
• 100x 1.4NA Oil  HC PL APO CS2

Detectors:

• 1 HyD
• 2 HyD SMD (Single Molecule Detection)
  
• 2 PMT
• 1 PMT Trans

FLIM/FCS/FCCS unit from Picoquant:

• PicoHarp 300 (TCSPC Module (Time Correlated Single Photon Counting system))
  
• Pulsed Diode Laser (PDL 800-B) 440 nm
  
• 4 Channel Detector Router (PHR 800)
  
• Software: SymPho Time 64 1.6

Software:

• Leica LAS X SP8 Version 1.0 (offline Leica LAS AF version 3.3 at Image processing station 2 in HPI F10.1)
  
• additional licences for STED, Matrix (with CAM) for HCS applications, Life data mode, Dye separation, 3D rendering
  
• Huygens Deconvolution
• FLIM/FCCS Software: SymPho Time 64 1.6 (Picoquant)

Key features:

• Stage: Motorized Märzhäusser stage with Leica Galvo stage
  
• Scanner: Tandem Scanner with Galvanometric scanning unit and Resonant scanning unit (8 kHz)
  
• Incubation: LIS box type temperatur incubation (Cube) with CO₂ (Fitting Ibidi, LabTeks and 35 mm dishes, Mikrotiteplates)

Fluorescence settings:

• For internal detection: Leica patented flexible filterfree detection within whole VIS range, spectral detection possible
  
• Fluorescence filter cubes for DAPI, FITC, TRITC for visual inspection