Enhanced imaging of fluorescently labeled specimens using a Leica TCS SP8 MP scanning confocal microscope
Lasers
- 405 nm for Dapi, Hoechst, CFP etc.
- 488 nm for FITC, Alexa Fluor 488, GFP etc.
- 514 nm for CFP
- 552 nm for rhodamine, Texas red, mRFP1, Alexa Fluor 546, 555, 568, 594 etc.
- 633 nm for Cy5 etc.
- Spectra-Physics Mai Tai DeepSee tunable ultrafast laser (680 - 1040 nm) for multi-photon excitation
Microscope
Leica TCS SP8 MP confocal microscope, with the following objectives
- HC PL Fluotar 10x 0.3 n.a.
- HC PL Apochromat 20x 0.75 n.a.
- IRAPO 25x 0.95 n.a. dipping lens for live cell and in-vivo imaging
- HC PL Apochromat 40x 1.30 n.a. oil
- IRAPO 40x 1.10 n.a. Corr water immersion
- HC PL Apochromat 63x 1.4 n.a. oil
System Resources
- Imaging of most standard fluorophores, in either single-photon or multi-photon modes.
- Simultaneous fluorescent and DIC imaging of most samples
- Heated, motorized stage for live cell imaging.
- Real time measurements of cell migration, ion flux and metabolic profiling (NADH, FP)
- Spectral imaging with up to 81 narrow wavelength bands (5 nm) over 380-785 nm for spectra-based experiments such as:
- Separation of fluorophores with closely-spaced emission peaks by linear un-mixing
- Generation of fluorophore-specific spectra
- Background subtraction
- Intensity based measurements of protein-protein interactions in live and fixed samples using FRET techniques
- Fluorescence correlation spectroscopy (FCS).
- Study of molecular dynamics in living cells
- Generation of diffusion coefficients
- Time-correlated single photo counting (TCSPC) using a Becker & Hickl spectroscopy system.
- Lifetime-based measurements of protein-protein interactions in fixed and living samples using FRET/FLIM techniques
- Software for FRET, FRAP and FLIM applications, ratiometric imaging, and complex time series experiments.
Additional Equipment
- Total Internal Reflection Fluorscence (TIRF) imaging on an inverted Olympus microscope (488 nm excitation).
- Single molecule bleaching and observations
- High axial resolution renderings of the features and events on the plasma membrane in living cells
We strive to provide dynamic imaging services and training for researchers and students including; experimental design, image acquisition, analysis (including statistics and figure generation) and potential installation of specialized equipment to meet each investigators research needs.
