Leica TCS SP8 MP Multiphoton/Confocal Microscope

Description:

Multiphoton microscopy (MPM) is an advanced fluorescence imaging technique for generating high-resolution optical sections from thick biological samples. While laser scanning confocal microscopy (LSCM) utilises a pinhole to remove out-of-focus lights from the image, MPM restricts the excitation volume to the laser focus using a pulsed long-wavelength laser. As the focus of the laser is the only location where photons are dense enough to excite the fluorophores, optical sectioning can be achieved without a pinhole. On the other hand, the use of excitation laser light in the infrared range is particularly suitable for live fluorescence imaging, for it would only cause minimal photocytotoxicity to live tissues and cells, as well as photobleaching to fluorophores when compared to the use of visible lasers in LSCM. Furthermore, infrared laser light can penetrate animal tissues with much less light scattering, making deep tissue imaging possible. The laser with continuously tuneable wavelengths also allows researchers to examine a wider range of fluorescent probes with diverse excitation spectra.
 

Specifications:

• DMi8 fully motorised inverted microscope body
• TCS SP8 confocal module
• Motorised XY-stage and focus drive
• SuperZ galvometric stage for rapid and high-precision Z-stacking
• Hardware active autofocus (Leica Adaptive Focus Control)
• Objectives (details):
• HC PL Apo 10×/0.40 CS2 Dry
• HC PL Apo 20×/0.75 CS2 Dry
• HC PL Apo 40×/0.85 Corr CS Dry
• HCX PL Apo 100×/0.95 Dry
• HC PL Apo 63×/1.20 Corr CS2 Water
• HC PL Apo 63×/1.40 CS2 Oil
• Solid-state laser lines for single-photon imaging:
• 405 nm
• 488 nm
• 514 nm
• 552 nm
• 638 nm
• Pulsed laser for multiphoton imaging:
• Mai Tai HP Ti:Sapphire compact laser system
• Tuneable from 690 to 1040 nm
• Pulse width: ~100 fs
• Average power: ~0.5 W at 690 nm; ~1.35 W at 710 nm; ~2.5 W at 800 nm; ~1.35 W at 920 nm; ~0.3 W at 1040 nm
• Internal detectors:
• One PMT detector
• Two HyD GaAsP detectors
• Imaging modes for HyD: standard, photon counting and Bright R
• Spectral detection from 380 to 800 nm
• External detectors:
• Two non-descanned HyD detectors (NDDs)
• HyD RLD 2-channel filter sets for FITC (525/50) and TxR (617/73)
• Scanners:
• FOV scanner (maximum image size: 8192×8192 pixels)
• Resonant scanner (maximum imaging speed: 28 fps at 512×512 pixels)
• Mercury metal-halide fibre optic illumination system with DAPI, FITC, Rhod filter cubes
• LAS X software with modules for FRET and FRAP
• Stage-top live-cell imaging chamber with temperature and CO₂ concentration control (Tokai Hit INU series)
• Perfusion system to be used in combination with the live-cell imaging chamber

Location: Y708c

Hourly rate: $183 (internal users); $366 (collaborators); $915 (external users)

Discount for long-duration bookings:

• Applicable to consecutive bookings of 8 hours or longer
• A 50% discount will be applied to any segments that fall within non-office hours (i.e., from 6 pm to 9 am on weekdays, and the whole days on Saturdays, Sundays and public holidays)
• Price adjustments, if any, will be made after the completion of the imaging session
• The ULS reserves the right to interpret these terms

Consumables:

• Marienfeld no. 1.5H 18 mm round coverglass ($148; 100 pcs)
• MatTek 35 mm no. 1.5 glass bottom culture dish ($190; 10 pcs)
• Nunc Lab-Tek II no. 1.5 8-well chambered coverglass ($435; 8 pcs)
• ProLong Diamond antifade mountant ($372; 2 mL)
• ProLong Live antifade reagent ($333; 1 mL)
• EMS 16% paraformaldehyde ($116; 10 mL)

Staff-in-charge: Dr Michael Yuen and Dr Alan Leung

Training: please contact Dr Michael Yuen (email: michael.yf.yuen@polyu.edu.hk / Tel.: 3400 8788) or Dr Alan Leung (email: alan.sl.leung@polyu.edu.hk / Tel.: 3400 8984) for arrangement.