The Leica SP8 is a high-end confocal system that is suitable for most standard confocal experiments, and additionally for superresolution microscopy. Superresolution is achieved through Stimulated Emission Depletion (STED), switching on powerful depletion lasers during acquisition to suppress fluorescence outside the point of interest. Optimally, this can boost resolution to ca. 60 nm.
For excitation, a pulsed White Light Laser (WLL) is available, which allows the selection of arbitrary excitation wavelengths between 470-670 nm. In addition, a UV diode, and an Argon laser for most common fluorophores are installed. At the detection site, PMTs and Hybrid (HyD) detectors are mounted. The HyDs make use of adjustable "time gating" which, coupled to the pulse rate of the WLL, allows filtering of signal contaminations originating from eg. reflections or autofluorescence.
Image acquisition in non-STED mode can be improved by using "HyVolution". This option allows deconvolution of images during the experiment, effectively increasing resolution ca. 2-fold on-the-fly. Acquired images can also be post-processed using the Huygens deconvolution software installed on the microscope PC.
An on-stage temperature incubator is available.
- 2 x HyD (time gated)
- 3 x PMT
- 1 x transmitted light
- WLL: 470-670 nm (pulsed 80 MHz)
- Diode: 405 nm
- Argon: 458, 476, 488, 496 and 514 nm
- STED-depletion: 592, 660, 775 nm.
- 20x multi / NA 0.75
- 40x water / NA 1.10
- 93x glycerol / NA 1.30 (STED suitable)
- 100x oil / NA 1.40 (STED suitable)