General

This system is

• An AMBIO-satellite at Charité Campus Benjamin Franklin department of clinical physiology/nutritional medicine

  Address:

  Hindenburgdamm 30, 12203 Berlin (Steglitz),

  main building, 5th floor, elevator 15, room 5505A

• Maintainance, training and assistance is done by Prof. Dr. Dorothee Günzel and PD Dr. Jörg Piontek
• Please contact us regarding labelling--especially if trying to push resolution limits (ambio@charite.de; dorothee.guenzel@charite.de; joerg.piontek@charite.de)

The Abberior Facility Line STED microscope is a high-end confocal and super resolution imaging platform:

• One STED laser (775 nm) and four excitation lasers (405, 485, 561, 640 nm)
• Patented adaptive illumination methods enable imaging with reduced photobleaching and phototoxicity
• Multi-channel, multi-position, time series, multi-ROI & tiling/stitching
• FRAP, FLIP, photoactivation, colocalization

  Olympus IX83 inverted basis:
• Fully motorized stage for accurate repositioning (IX3-SSU ultrasonic XY-scanning stage for IX83)
• Z-drift compensation (Olympus IX3-ZDC-12 z-drift compensation unit for steady confocal/STED imaging)

  Imaging Modes

  STED plus confocal:

• Two main STED (stimulated emission depletion) -channels: StarRed/Atto647N/SiR (with excitation at 640 nm) & StarOrange/Alexa Fluor 594 (with excitation at 561 nm)
• An optional third less efficient STED channel, for instance for STAR 460L (long Stokes-shift dye) with excitation at 485 nm
• Plus up to two confocal channels: DAPI & Alexa Fluor 488/GFP (or similar fluorophores)
• 2D-STED: Resolution XY > 40 nm, Z > 500 nm including Z-stacks for 3D-imaging
• 3D-STED: Resolution down to XYZ > 100 nm including Z-stacks for 3D-imaging

Confocal only:

• Resolution down to XY > 220 nm, Z > 500 nm
• 4 channels with fluorophores similar to DAPI, Alexa Fluor 488/GFP, Alexa Fluor 555/Alexa Fluor 594 & Alexa Fluor 647/Cy5

Widefield EPI fluorescence:

• 4 channels with fluorophores similar to DAPI, Alexa Fluor 488/GFP, Alexa Fluor 555, Alexa Fluor 594, Alexa Fluor 647/Cy5

Excitation Lasers

• 405 nm (cw)
• 485 nm (pulsed)
• 561 nm (pulsed)
• 640 nm (pulsed)

STED-Laser

• 775 nm (high-power variant, pulsed)

Detectors

• Three ultra-sensitive single-photon-counting APDs with tunable detection bands between 400 and 800 nm
• Matrix detector: MATRIX array detector with more than 20 avalanche photodiodes in single photon counting mode
• Four-color widefield module for epifluorescence with camera port on Olympus (405/470/590/635)

Objectives

Special

• Adaptive illumination via RESCue, DyMIN, and MINFIELD

• RAYSHAPE dynamic aberration correction
• EASY3D STED
• Confocal & STED autofocus
• Spectral RAINBOW detection
• Autoalignment
• Software Base Package Imspector for Facility Line