The confocal STED microscope is mainly used for high-resolution (100 nm) imaging of specific parts of the cells, such as the surface membrane, labelled proteins, etc. The white excitation laser and the three depletion lasers allow the use of a wider range of fluorescent labels and multiplexed staining; the 405 nm laser adds the possibility of using a fourth label for the nucleus and also photoconversion of specific samples. This microscope can also be used for standard confocal microscopy. The range of fluorescent specimens that can be used is even wider than with STED. The quality of the images obtained is greatly improved by gating. The pulsed laser and hybrid detectors remove a large part of the non-specific light (reflection, scattering).
The tandem scanner allows the measurement of local 3D (xyt) and 4D (xyzt) calcium signals using calcium indicators. Other speed-intensive techniques can also be performed. For example, spectral separation in 3D samples can be performed and the excitation-emission spectra of the sample displayed.
Leica TCM SP8 STED 3X Microscope STED Configuration:
- DMi8 inverted microscope
- AOBS
- Standard objectives 63x water and oil, special objective STED WHITE 100x
- Tandem scanner 8 kHz
- 2x HyD, 2x PMT, 1x TLD setectors
- Gated STED , 3D STED
- SuperZ galvo for fast 3D imaging
- Laser 405 nm
- Pulsed white laser WLL2 – 470 – 670 nm
- Laser depletion for STED: 592 nm, 660 nm, pulsed laser 775 nm
Contact: Mgr. Zuzana Ševčíková Tomášková, PhD.
Support: ITMS 26230120006