Leica SP8 Confocal, STED, FLIM System

CONFOCAL/STED/FLIM Imaging System, BL-2

Leica SP8 confocal microscope for live cell imaging with high-speed, multi-spectral, super-sensitivity, super-resolution, and fluorescence lifetime capability. This instrument is in a Biosafety Level 2 (BSL-2) imaging lab.  

Instructions: How to get access to the BL-2 lab

Training Policy: Leica SP8/STED/FLIM Training Policy

 

Available objectives include:

  • HC PL APO 10x/0.40 CS2
  • HC PL APO 20x/0.75 IMM CORR CS2 - multi-immersion objective (oil, glycerol, water)
  • HC PL APO 40x/1.10 W motCORR CS2 - water immersion, motorized aberration correction
  • HC PL APO 63x/1.30 GLYC CORR CS2 - Glycerol immersion for samples mounted in glycerol-based mounting media
  •  HC PL APO 100x/1.40 OIL for STED superresolution

 

FALCON - Fluorescence Lifetime Imaging module - follow fast molecular interactions via FLIM-FRET (Förster Resonance Energy Transfer).

AOBS and WLL (470 nm-670 nm) white light laser source of the Leica TCS SP8 X perfectly matches the wavelength of any fluorophore in the visible spectrum. Up to eight excitation lines can be used simultaneously. By tuning both excitation and detection, complete excitation emission spectra can be acquired.  A pulsed 440 nm laser is coupled with the system for FRET or FLIM-FRET, and 405 nm laser line is included with the system for UV imaging.

LightGate is a filter-free method to remove unwanted short-lifetime signal, such as tissue autofluorescence or unwanted reflection.

Lightning Deconvolution resolves structures down to 120 nm. GPU-based real-time processing.

Hybrid Detectors: HyD SMD  with superior sensitivity and single photon counting The system includes 3 hybrid detectors.

APD (Avalanche Photodiode Detector) for excellent sensitivity in red/far red region of the spectrum to handle new NIR probes.

STED

 STED 775 tunable and direct nanoscopy in x,y (2D) with resolution down to ~40 nm.  Designed for dyes like Abberior STAR 635 or Alexa Fluor 647. A second color is possible with dyes excited around 590 nm.    

Resonant Scanning mode (Tandem Scanner) for high speed scanning and less phototoxicity to live cells. Scan at frequencies of up to 12 kHz, resulting in about 40 fps (512 x 512 px) or 428 fps (512 x 16 px). Acquire 4D stacks up to 50% faster than comparable systems by combining them with the SuperZ GalvoFlow.

Live Cell Imaging - with Adaptive Focus Control (AFC) to automatically maintain focus. Tokai Hit stage top incubator for long term live cell imaging maintains humidity, temperature and CO2.

Navigator Software ModuleCreate fast overviews of samples,  then set up high resolution image acquisition automatically using templates for slides, dishes and multiwell plates. Allows for high throughput imaging, image tiling and stitching.