Stellaris Laser Scanning Confocal​​​​

Description

This is a super-resolution laser-scanning confocal outfitted with 3 STED depletion lasers. It is capable of standard laser-scanning confocal microscopy, STED, tau-STED, and FLIM. With the White-Light Laser (WLL) it can be used to image a wide variety of fluorophores and is capable of separating fluorophores spectrally close.

Location

Weill Neurosciences Building
Room 451J

Contact

Caroline Mrejen, PhD
​[email protected]

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Hardware

  • Leica DMi8 Inverted Microscope
  • Leica Adaptive Focus Control (AFC)
  • Scanning stage SCAN plus inverted motorized stage
  • Z-Piezo stage with 500um travel (WieneckeSinske WSBPiezo)
  • Water Immersion micro dispenser
  • 8kHz Tandem Scanner STELLARIS 8 (galvo and resonant)
  • HyD detectors
    • HyD S (pos 1 and pos 3)
    • HyD X (pos 2 and pos 4)
    • HyD R (pos 5)

Objectives

Installed objectives
  • HC PL APO 86x/1.20 W motCORR STED W (Calibrated for Pos 2, occupies 2 positions)
  • HC PL APO 10x/0.4 NA CS2
  • HC PL APO 20x/0.75 NA CS2
  • HC PL APO 40x/1.30 Oil CS2
  • HC PL APO 100x/1.40 OIL STED W (Calibrated for Pos 6)
Objectives stored off the microscope
  • HC PL APO 63x/1.40 OIL CS2 (Move to SD)
  • HC FLUOTAR L 25x/0.95 W VISIR
  • HC PL APO 93x/1.30 GLYC motC STED W (Calibrated for Pos 4, occupies 2 positions)
  • HC PL APO 40x/0.95 CORR

Light Source

  • Leica LED 3 (390-680nm) with 3mm LLG

Excitation Filters

  • Widefield
    • LED 405 filter cube
    • CFP/YFP filter cube
    • GFP filter cube
    • Rhod LP filter cube
    • DA/FI/TX filter cube

Lasers

  • 405nm
  • White Light Laser  (pulsed, 440 to 790 nm)
  • 592 nm STED depletion laser
  • 660 nm STED depletion laser
  • 775 nm STED depletion laser

Laser Notch Filters

Fluorifier Disc 1
  • Empty
  • NF 448/514/561
  • NF 488/561/730
  • NF 448/514
  • NF 488/561/638
Fluorifier Disc 2
  • Empty
  • NF 685
  • NF 638
  • NF 488/638
  • NF 775
  • NF 660

Detector

Detector HyD S HyD X HyD R
Dead Times <1.3 ns <1.6 ns <1.6 n
Peak PDE > 58% @ 500nm > 46% @ 500nm > 26% @ 635 nm
Photon count rates for STED 300 Mcps 300 Mcps 300 Mcps
Tau STED using power counting mode   100 Mcps 100 Mcps