Leica Sp8 Lightning Single Photon
The Biology Department Leica SP8 (Spectral Detector) Lightning LSCM was installed in December of 2018. It is built upon a Leica DMi8 fully motorized inverted microscope and offers a resonant scanner in addition to the conventional galvanometer scanner (x2y Tandem Scanner); a piezo driven stage z drive; two ultra-sensitive HyD detectors plus two conventional PMT detectors; and a transmitted light detector. Channel separation for dyes is achieved via a fully-tunable prism-based spectral detector to provide unparalleled control of wavelength transmission. The piezo drive provides ultrafast Z scan acquisition, and the resonant scanner can scan at 8000 Hz for even the fastest of live cell imaging requirements. The instrument has 5 lasers running at 405 to 638 nm, along with 4 objectives ranging from 10X to 63X. Three modular stage inserts accommodate slides, petri dishes, and multiwell plates. Image acquisition and processing is run by Leica LAS X software and includes modules for Tilescanning on the fly (Navigator), FRET, FRAP, 3D viewing and analysis, and Leica’s proprietary Adaptive Deconvolution package (Lightning). The software is extremely user-friendly, walking users through complicated experimental designs quickly and easily.
Leica DMi8 inverted microscope with Broadband EL6000 mercury halide lightsource and 3 filter sets for DAPI, FITC and Rhodamine. White transmitted light source with Brightfield, Polarized and DIC optics. The stage is fully motorized with standard and Super Z galvo Z drives. There are three piezo-driven adaptor plates for microscope slides, petri dishes, and multiwell plates, as well as a standard Z-drive adaptor plate for slides and multiwell plates.
Objective | Magnification | Numerical Aperture |
HC PL APO CS2 |
10X dry |
0.40 |
HC PL APO CS2 |
20X multi immersion |
0.75 |
HC PL APO CS2 |
40X oil immersion |
1.30 |
HC PL APO CS2 |
63X oil immersion |
1.40 |
Lasers | Power | Power at Objective |
405 nm DMOD compact | < 60 mW | < 5 mW |
488 nm DPSS | < 12 mW | < 3 mW |
514 nm DPSS | < 12 mW | < 3 mW |
552 nm DPSS | < 12 mW | < 3 mW |
638 nm DPSS | < 12 mW | < 3 mW |
SP8 LIAchroics Compact RGB x2y Tandem scanner: 8000 Hz resonant scanner + 10-1800 Hz galvanometric scanner.
Two PMT confocal detectors + Two HyD GaAsP-Detectors for gated imaging. One brightfield transmitted light detector.
LAS X software with modules:
Module | Uses |
Navigator |
GUI for point scanning and Tiled images |
Dye Finder |
Dye setup wizard for multicolor imaging |
3D visualization |
3D viewer, processor and analysis wizard for Z stacks |
MicroLab |
Wizards for FRAP, FLIP and FRET |
FRAP Zoomer |
FRAP beam control for enhanced photobleaching |
Lightning |
Adaptive Deconvolution software |
Applications:
FRAP, FRET, photoactivation, 3D rendering and analysis, Timelapse, Deconvolution, colocalization, point scanning, Tiling, Z plane acquisition.
Output:
.lei, .tiff, .jpg, .avi, .mov, .mpeg4