The Leica TCS SP8 with single molecule detection (SMD) and multiphoton laser (MP) is our new state of the art confocal microscope system.

With the Leica SP8, you can do routine imaging, high-speed live cell imaging, single photon detection and high content screening. It is equipped with several visible lasers and an infrared pulsed laser for multiphoton (MP) fluorescence and second harmonic signal generation (SHG). The hardware and software from PicoQuant, needed for fluorescence lifetime imaging (FLIM)and fluorescence correlation spectroscopy (FCS), is fully integrated with the Leica TCS SP8.

The microscope

Leica DMI6000 inverted and automated microscope with motorized XY-stage, touch control panel and adaptive focus control (AFC). Observation modes in addition to confocal imaging are brightfield, differential interference contrast (DIC) and epifluorescence

Focus drives

• Motorized objective nosepiece (9 mm travel range in Z)
• SuperZ galvanometer stage (1500 um travel range in Z) for higher speed and accuracy.

Scanners

• Tandem scanner (switchable between galvo and resonant 8kHz mode), up to 29 frames /sec at 512 x 512 pixels image format can be achieved.

Objectives

• The 20x immersion objective is for water, glycerine and immersion oil
• The 63x water objective have a motorized correction collar and can be used with the micro dispenser.

Incubation system and accessories

1. Large incubator covering the entire microscope, for warm air incubation and CO2-control.
2. Incubator-kit for Super Z Galvo stage, including different inserts and mask sets for the stage.
3. Micro dispenser for automated supply of water immersion during long term experiments.
4. Hardware-based focus stabilizer.

Condenser top

• NA 0.55 (S28) air, working distance 0.28 mm
• NA 0.90 (S1) air, working distance 1 mm
• NA 1.40 (S1) oil, working distance 1 mm

Lamps

Halogen: 12V - 100W.

Mercury shirt arc lamp HXP 120W 

Lasers

• The MP laser is the TiSp-laser Chameleon Vision-S from Coherent
• An acousto-optical tunable filter (AOTF) controls the intensity of the visible excitation laser light.
• An electro-optical modulator (EOM) is used to control the intensity of the MP laser.
• An acousto-optical beam splitter (AOBS) is used to split excitation light and emission light instead of filter blocks.

Scanners

Conventional scanner: 10-1400 Hz (2-3 frames/sec at 512 x 512 pixels image format)

Resonant scanner: 8000 Hz (25 frames/sec at 512 x 512 pixels image format)

Detectors

Internal detectors in reflection mode (with pinhole) with freely tunable emission bands in the range 400 – 800 nm.

• 2 x HyD (GaAsP detectors)
• 2 x PMT (standard photomultiplier tube)

External detectors with customizable filter cubes

• 2 x HyD (GaAsP detectors, without pinhole) in reflection mode. Used for MP imaging and SHG
• 2 x PMT (without pinhole) for photon detection in transmission mode.
• 2 x SPAD (single photon avalanche diodes from PicoQuant, with pinhole) in reflection mode for SMD: FLIM and FCS. Filter cube: FITC/TRITC

Light for epi-fluorescence

• External EL 6000 (HXP 120 W): fast shutter, fiber-optics and bulb lifetime 2000h
• Halogen lamp: 12V – 100W

Filter cubes for epi-fluorescence

Software on the SP8 computer

Leica LASX352 with software modules

1. FRAP Wizard
2. FRAP XT Wizard
3. FRET AB Wizard
4. FRET SE Wizard
5. FLIM Wizard
6. FCS Wizard
7. Matrix Screener
8. Live Data Mode
9. Advanced Mark & Find

Software on the SMD computer

SymPhoTime 64 bit version from PicoQuant.