Leica TCS SP8 STED and MP

The Leica SP8 spectral confocal microscope is equipped with three high sensitivity, low-noise hybrid detectors capable of photon counting and two conventional PMT detectors. Sample excitation can be accomplished by multiple lasers including a pulsed white light laser tunable from 470 to 670 nm that allows selection of the optimal excitation wavelength; the LightGate panchromatic electronic barrier filter allows gating of the detectors to avoid collecting signal during the laser pulse so that only pure emission signals are collected. The system also has conventional fixed laser excitation lines at 405, 458, 476, 488, 496, and 514 nm. For two-photon imaging, including FLIM-FRET, the system has a MaiTai DeepSee Ti-Sapphire ultrafast laser (tunable between 680–1060 nm). The SP8 is equipped with both a conventional point scanner for work requiring the highest sensitivity and a resonant scanner for live cell imaging (28 fps at 512x512). An environmental chamber for temperature and CO2 control during prolonged live-cell imaging experiments is available. Complex experimental designs, 3D image tiling and 3D rendering and measurements are possible with the LAS AF Leica software.

The Leica confocal microscope has been upgraded with a 3D STED super-resolution system. This system can achieve a resolution of 50 nm or less. Three depletion lasers (592, 660 and 775 nm) enable multicolor detection of a wide range of fluorophores. Use of the Huygens deconvolution software programs allows for further improved resolution.

Download a PDF describing the basics of STED technology and sample preparation.

The following objectives are available for this system:

Mag.Objective DescriptionNAImmersion MediumWorking distance (mm)
10xHC PL APO CS0.40Air2.20
20xHC PL APO CS0.70Air0.59
20xHC PL APO IMM CORR0.75Water, glycerol, or oil0.68
25xHCX IRAPO L0.95Water2.40
40xHC PL APO CS21.10Water0.65
40xHC PL APO CS21.30Oil0.24
63xHC PL APO CS21.40Oil0.14
63xHC PL APO CORR CS21.30Glycerol0.30
63xHC PL APO CORR CS21.20Water0.30
100xHCX PL APO CORR CS1.44Oil0.10
100xHC PL APO STED WHITE1.40Oil0.13