Integrated Light Microscopy Core

2-photon Microscopy

Deep tissue imaging
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Hands-on Microscopes

To access a microscope, click the New User Training button above and work through our training checklist. Trained users have 24/7 access to the facility and are given permission to schedule their own microscope sessions through our online scheduling software.
Leica SP5 2-photon Laser Scanning Confocal

Overview: This laser scanning confocal system features software selectable conventional or high-speed resonance scanner galvanometer system with three (chilled, high sensitivity) internal PMT detectors (spectral) and two external (NDD) PMT detectors. Six visible laser lines and a tunable NIR pulsed laser (Spectra Physics Mai Tai broadband 710-990nm) provide excitation. The system features turnkey operation and full software control in addition to the features listed below.

Location: KCBD 1250B

Training Contact: Lorraine Horwitz

Fluorophores this microscope can image:

  • Blue (ex: DAPI, Alexa 405)
  • Cyan (ex: CFP)
  • Green (ex: GFP, Alexa 488)
  • Yellow (ex: mCitrine, YFP)
  • Orange (ex: Cy3, Alexa 543)
  • Near Red (ex: Texas Red, Alexa 594)
  • Far Red (ex: Cy5, Alexa 647)
  • Near IR (possible but not ideal, ex: Alexa 700)
  • 2-photon imaging of Blue, Cyan, Green, Yellow, Orange

Excitation Light Source(s):

  • near UV laser at 405nm
  • 5 line Argon laser at 458, 476, 488, 496 and 514nm
  • HeNe laser at 561nm
  • HeNe laser at 594nm
  • HeNe laser at 633nm
  • MaiTai laser tunable from 710 - 990nm

Emission Detection:

  • Custom emission detection of any wavelength range between 410nm and 800nm
  • 3 chilled photon multiplier tubes (PMTs)
  • 2 hybrid GaAsP/PMT detectors (HyDs)
  • 2 non-descanned detectors (NDDs)
  • Transmitted light detector
  • 8-, 12-, or 16-bit grayscale output

Objectives (with additional optical zoom possible):

  • 20x / NA 0.8 dry
  • 40x / 0.8 water
  • 63x / 0.9 water
  • 63x / 1.4 oil
  • 100x / 1.46 oil
  • 25x / NA 0.95 water (must be installed)
  • Objectives listed under the SP5 STED description (must be installed)

Sample Chamber:

  • Custom fit full incubation jacket (clear) for increased thermal stability during live cell experiments
  • Inverted platform for imaging of slides or dishes
  • LSM objective inverter device (allows system to be used as if it were an upright microscope platform)
  • Automated XY stage for tiling / multipoint scanning
  • "SuperZ" galvo focusing stage, 1.5 mm range. Demonstrated capability of three cell volumes per second (12 x 1-mm optical slices each)
  • Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

  • FRET, deconvolution, and 3D software wizards
  • Automated notch filters
  • Spectra Physics Mai Tai broadband Ti:sapphire laser with 10W pump (tunable from 710 to 990 nm) (up to 2W output at 810 nm)
  • Dual scanning galvanometer mirrors (max 16,000 Hz scan rate) 25 images/sec at 512x512; strip scans to 333 fps (4 channels). Standard scanner with beam park, bleaching, imaging to 8k x 8k (64 megapixels) per channel.
  • AOTF control of visible laser lines, EOM and ND filter attenuation of NIR
  • Excitation and emission scanning capability with linear unmixing
  • Automatically optimized confocal pinhole apertures
  • Transmitted detector with LWD and oil lenses and filters suited for SHG imaging