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STED Microscopy

STED Super-Resolution Microscope

Since 2017  MPFI has utilized the Abberior Instruments Expert-Line 3D-STED, enabling users to record super-resolved fluorescent images with comparative ease. By design, the AIA STED can operate as a fully functional confocal microscope, and as such can image any typical fluorophore in the visible and near-IR spectrum. Furthermore, by activating the depletion (STED) beams, the resolution of the fluorescent images can be increased by up to an order of magnitude in x,y or z. In general, any good confocal sample can be imaged on the AIA STED nanoscope, although some fluorophores vastly outperform others in terms of brightness, photostability and achievable resolution.

 

Services

  • Fixated or live sample imaging (at room temperature or heated using a dedicated sample chamber and objective heater)
  • Labeling with fluorescent proteins, organic dyes and other fluorescent markers possible
  • Deep imaging inside thick tissue samples (<100µm works best)
  • Subdiffraction spatial resolution between 30nm–50nm (2D) and 90nm–110nm (3D)
  • Two STED lasers for imaging in the blue/green (595nm STED) and the orange/far-red spectral range (775nm STED)
  • Simultaneous, multi-channel recording of up to 4 channels by pulse-, pixel-, line-, or frame-interleaved acquisition
  • Multi-color imaging of up to 4 colors with STED resolution (or confocal, or a mixture of both)
  • Image-sectioning (for 3D-stacks) and time-lapse imaging
  • Photon-counting detection modes (FLIM, etc.)
  • RESCue mode imaging for reduced photobleaching and -toxicity

 

Equipment

Abberior Instruments Expert-Line 3D-STED
Inverted stage; 4 excitation lasers @405nm, 485nm, 561nm & 640nm; 2 pulsed STED lasers @595nm & 775nm; easy3D STED depletion module; beam-scanner; motorized pinhole; four detection channels (APDs) between 425nm – 725nm; time-gated detection; single-photon counting; RESCue; objectives: 10x/0.25 air, 40x/1.3 oil, 63x/1.2 water, 100x/1.4 oil)

The AIA STED is fully equipped to image:

  • Both fixated and live samples
  • Cultured cells and tissue samples (thickness <100µm works best)
  • At room temperature or heated using a dedicated chamber and objective heater
  • Fluorescent proteins, organic dyes and other fluorescent markers

 

Contact

Nicolai Urban
Microscopy Specialist
(561) 972-9175
nicolai.urban@mpfi.org