Leica SP8 MP

They can be used for simultaneous two channel MP acquisition at optimal excitation wavelengths for different fluorophores. Both systems are equipped with sensitive Leica Hybrid detectors (HyD SP GaAsP, 2x more sensitive than conventional PMTs). Find more details about the hybrid detectors external pageherecall_made.

MP Illumination (Multiphoton lasers):

• Mai Tai XF (Spectra-Physics), tunable from 720–950 nm, additional DeepSee (prechirp) unit, intensity regulated vial EOM, ROI scan possibility, details can be found external pageherecall_made
  
• InSight DeepSee (Spectra-Physics), @ this setup tunable from 950–1300 nm, average power, intensity regulated via attenuator wheel, details can be found external pageherecall_made

Inverted system

• Multiphoton and full confocal acquisition
• Fast acquisitions with 8 kHz resonant scanner
• Fast Z-scan aquisition with Z galvo stage (galvo flow option)
• Hardware autofocus option (Adaptive Focus Control) with confocal acquisitions
• Tile scan and multi position experiments with motorized stage
  
• Spectral detection with Lamda scans (emission and excitation)
• Automated water immersion supply unit for 40x Water objective

Microscope:

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

Illumination:

• Laser (CW) unit for confocal acquisition (AOBS system): 405, 458, 477, 488, 496, 514, 561, 633 nm
• MP lasers (see above) for MP acquisitions
  

Objectives:

• 10x 0.3NA PH1 HC PL FLUOTAR
• 20x 0.7NA PH2 HC PLAN APO
  
• 20x 0.7NA, Oil/W/Glyc, HC PLAN APO
• 25x 0.95NA L Water HCX IRAPO
• 40x 1.1NA Water HC PL IRAPO CORR
• 63x 1.4NA Oil CS2 HC PL APO

Detectors:

• Internal detection: 1 conventional PMT Detector and 2  Leica Hybrid detectors
  
• External detection in backward direction (reflection): 2 channel NDD detector unit with 2 Leica Hybrid detectors and Filtercube / Shortpass filter ST 680 nm in front (ST815 for far red detection pending)
  
• External detection in forward direction (transmission): 2 channel NDD detector unit with 2 conventional PMTs and filtercube (transmission channel with 1 conventional PMT on demand)
  

Software:

• Leica LASX SP8 (offline Leica LASX on VM and image processing station 1, 2 and 5)
  
• additional licences for Matrix (with CAM), live data mode, dye separation
  

Key features:

• Stage: Motorized Märzhäusser stage
  
• Scanner: Tandem Scanner with Galvanometric scanning unit and Resonant scanning unit (8 kHz)
  
• Incubation: LIS box type temperatur incubation (Cube)

Fluorescence settings:

• For internal detection: flexible filterfree detection within whole VIS range, spectral detection possible
• Possible filter combinations for external detection:
  1. FITC (525/50)/TRITC (585/40)
  2. FITC/TexasRed (617/70)
  3. DAPI/FITC, CFP/YFP
     
  4. on demand: DAPI or CFP in combination with TRITC or TexasRed)
  5. far red detection                     
     

Upright system

• Mainly Multiphoton acquisition
• Limited confocal acquisition possible with a solid state 488 nm laser and conventional PMT detection
• Spectral detection with Lamda scans (emission and excitation)
  

Microscope:

Leica DMI6000 CFS with red/green epifluorescence illumination for visual observation through eyepiece

Illumination:

• solid state 488 nm laser for confocal acquisition
• MP lasers (see above) for MP acquisitions

Objectives:

• 25x 0.95NA L Water HCX IRAPO 
  

Detectors:

• 1 internal conventional PMT Detector for confocal (and MP) acquisition
  
• 2 channel external NDD detectors unit in reflection direction: 2 Leica Hybrid  detectors with Filtercube and Shortpass filter in front
  

Software:

• Leica LASX AF SP8

Key features:

• Stage: Manual stage or custom made setup for animal fixation
• Scanner: Galvanometric scanning unit
• Incubation: LIS box type temperatur incubation (Cube) also functioning as a laser safety housing

Fluorescence settings:

• For internal detection: flexible filterfree detection within whole VIS range, spectral detection possible
  
• Possible filter combinations for external detection:
  1. FITC (525/50)/TRITC (585/40)
  2. FITC/TexasRed (617/70)
  3. DAPI/FITC, CFP/YFP
     
  4. on demand: DAPI or CFP in combination with TRITC or TexasRed)
  5. far red detection