LEICA SP8 STED3X SMD/FCS Confocal Microscope

LEICA SP8 STED3X SMD/FCS Confocal Microscope
Objectives

10x/NA 0.40 air
20x/NA 0.75 multi-immersion (air, oil, glycerol)
40x/NA 1.10 water
63x/NA 1.20 water
63x/NA 1.40 oil
100x/NA 1.40 oil

Laser lines

Diode CW – 405nm
Diode Pulsed – 440nm
Argon – 458, 476, 488 and 514nm laser lines
White Light Laser Pulsed tunable from 470nm to 690nm

Depletion Laser lines

592nm CW
660nm CW
775nm Pulsed

Detectors

2 standard (GaAs) PMTs
2 high sensitivity (HyD) PMTs
1 PMT for transmitted light

Additional techniques

The system provides a STED 3X module for STimulation Emission Depletion super-resolution with 3 depletion lines. The module can improve both the lateral and axial resolutions beyond the Abbe’s limit of standard fluorescence microscopy.

Thanks to the tunable excitation, the system can reconstruct both the emission and excitation profile in a so-called Λλ mode. This allows a characterization of autofluorescence properties of a sample and the spectral unmixing of dyes with strong overlap.

In addition to standard galvanometric scanners, a resonant scanner coupled with a piezoelectric z-stage allows very fast recording (30fps and higher) of bright stainings in live cells. An Autofocus module guarantees the stability of the focus position.

The system also provides Single Molecule Detection (SMD) capabilities, with two Hybrid photon counting detectors and dedicated acquisition hardware. This module has a very broad range of applications, from FLIM/FRET experiments to microenvironment quantification of pH and ion concentrations in live cells, to the binding and conformational properties of molecules in FCS modalities.