Technical Specifications

CGRB and EHSC Confocal Microscope Facility

Zeiss LSM510 META with Axiovert 200 motorized microscope with version 3.2 LSM software.

1. Inverted Microscope and Objectives
   1. Array of objectives for different needs (e.g., long working distance, dry and oil immersion)
   2. DIC optics

   4x/0.1 dry	100 mm
   10x/0.3 dry	130 mm
   25x/0.8ImmKorr(oil, water or glycerin)	130 mm
   40x/0.75 dry	500 mm
   40x/1.3Oil DIC	200 mm
   40x/0.80 Water	3.2 mm
   63x/1.4 Oil DIC	180 mm
   63x/0.95 water	2.0 mm

2. Laser Module
   1. Four lasers (Diode 405, Argon, HeNe543, HeNe633), seven laser lines (405 nm, 458 nm, 477 nm, 488 nm, 514 nm, 543 nm, 633 nm) for a variety of fluorescent probes
   2. Near UV laser (405 nm) for photo-activatable GFP
3. Features (Scanning and Electronics and Software)
   1. META
      • META module (32 PMTs with optical grating) detects the full spectra of multiple fluorophores simultaneously.
      • META controlled by Digital Signal Processors (DSP) for fast, flexible synchronization of the scanners.
      • Ability to establish spectral emission characteristics for distinct fluorophores into different channels.
      • Linear unmixing to separate highly overlapping spectra (even GFP and FITC) (e.g., for extraction of autofluorescence).
   2. Two regular PMT detectors
   3. Live cell incubation chamber (physiological chamber and perfusion chamber) with objective heater).
   4. Physiology software.
   • Precise laser controls for Fluorescence Recovery After Photobleaching (FRAP)
   • Fluorescent Resonant Energy Transfer (FRET) capability (e.g., for the CFP YFP pair) for studying protein-protein interaction in live cells.
   5. Motorized X-Y scanning stage for control of large and multiple specimens.
   6. Stitch Art software: 'Stitching' software for analysis of large (multiple image) specimens (to be obtained in the next 6 months.
   7. Multi-time software: for use in automating repetitive tasks such as repeating Z-stacks over time, scanning the same set of locations over time.
   8. Z-stack and l-stack with spectral distributions of fluorescent emissions.
   9. Simple 3D software with capacity to do 3D reconstruction and rotation.
   10. User-friendly interface - several simple 'walk-through' protocols for beginners, expert users have extensive options for changing parameters and writing macros.
   
   
   

   Support from EHSC/Tamara Fraley/Jeff Greenwood's lab 737-3198 and CGRB/Anne-Marie Girard/CGRB Core Labs 737-3413