Leica SP8 Confocal, STED, FLIM System
|$65/Hour||TAMU Users & Federal Government Agency Users|
|$55/Hour||Off-Peak & Automated Runs
(TAMU Users & Federal Government Users only)
5:00 p.m. - 8:00 a.m.
Gallery Image: Drosophila gut stem cells expressing chimeric epidermal growth factor receptor taken on the Leica SP8 confocal by Dr. Robert Fuentes and the sample prepared by Dr. Mohamed Mlih.
The Leica SP8 confocal microscope is for live cell imaging with high-speed, multi-spectral, super-sensitivity, super-resolution, and fluorescence lifetime capability.
- Live Cell Imaging with Adaptive Focus Control (AFC) to automatically maintain focus in time-lapse experiments.
- Navigator Software Module creates a fast overview of samples.
- Lightning GPU-based Deconvolution resolves structures down to 120nm.
- Resonant Scanning Mode for high speed scanning and less phototoxicity to live cells. Scan at speeds up to 40 fps (512 x 512px) or 428 fps (512 x 16px).
- Gating to remove unwanted short-lifetime signal such as tissue autofluorescence or unwanted reflection.
- FALCON-Fluorescence lifetime imaging module-follow fast molecular interactions via FLIM-FRET (Forster Resonance Energy Transfer). Phasor Analysis
- Superrsesolution with STED (Stimulated Emission Depletion) with XY resolution down to ~40 nm, tau-STED mode (utilizes FLIM for increased resolution)
Imaging Modes: Fluorescence spectral imaging, STED superresolution, FLIM, Brightfield, and DIC
- 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
- 405 nm CW laser, for dyes excited in UV and violet (e.g., DAPI, Hoechst dyes)
- 440 nm pulsed laser for fluorescence and FLIM, FLIM-FRET (e.g., CFP, CFP-YFP FRET)
- WLL (470nm-670nm), white light pulsed laser. Choose up to eight emission lines anywhere in the range. Allows acquisition of excitation and emission spectra.
- 775 nm pulsed depletion laser for STED superresolution. Allows x,y (2D) resolution down to ~40nm. This is designed for dyes like Abberior STAR 635 or Alexa Fluor 647. A second color is possible with dyes excited around 590nm.
- Two standard PMT detectors
- Three hybrid detectors HyD SMD with superior sensitivity and single photon counting.
- Two APD (Avalanche Photodiode Detector) for excellent sensitivity in the red/far red region of the spectrum to handle new NIR probes.
- Standard motorized stage, Galvo insert for fast focusing
- OkoLab enclosure and stage top gas chamber for long-term, live cell imaging maintains humidity, temperature and CO₂.
Software: LAS X software modules for multi-dimensional, STED and tau-STED, FCS, FRAP, FLIM imaging with Phasor analysis.
For imaging services, training or other questions
Instructions, FAQs and How-to documents:
- The instrument is in an access-controlled BSL-2 laboratory: How to get access to the BL-2 lab
- How is the training structured, how long does it take? Leica_SP8-STED-FLIM_Training_Policy.pdf
- TAMU MIC Leica SP8 User Guide – Startup/shutdown procedures, Instrument settings, Setting up multi-channel image acquisition, LIGHTNING deconvolution, ..
- Which fluorescent dyes, mounting media and slides/Imaging chambers should be used?
- LAS X Core confocal and FLIM viewer software: Download here – https://webshare.leica-microsystems.com/latest/core/confocal/
- Fluorescence Spectra Viewer: https://public.brain.mpg.de/shiny/apps/SpectraViewer/
- Leica guidelines to STED sample preparation: choosing fluorochromes, mounting media, … https://www.leica-microsystems.com/science-lab/the-guide-to-sted-sample-preparation/
- Leica LAS X tips and tutorials – Leica Microsystems YouTube channel – Instructional videos on many aspects of image acquisition in LAS X (Navigator, multi-well plate imaging, autofocus, flat field correction, File management, opening LAS X files in FIJI/ImageJ, ..).
Video: Adjusting the correction collar on the 20x multi-immersion objective (or the 63x glycerol immersion objective), using the endoscope camera installed on the system.