Imaging Facility

Super resolution (STED)

Achucarro's Leica TCS STED CW SP8 super-resolution confocal microscope

Our Imaging Unit is equipped with a Leica TCS STED CW SP8 super-resolution microscope. This confocal microscope allows exploring details of samples (in our case, brain tissue), below the limits of diffraction barrier with a purely optical approach, even on living samples in real time. Its technology allows super-resolution optical sections, which can penetrate the samples, and allow the observation of dynamic cellular processes at the nanoscale. It is equipped with a white laser for a more flexible and precise excitation and a super-resolution system (STED extended with Gated STED) that solves structural details in the range of 50-80 nm in samples with double fluorescent labeling.

Main technical specifications

  • Inverted Leica DMI 6000B microscope.
  • Objectives: 10x Air, 20x Air, 40x Oil, 63x Oil, 100x Oil (g-STED).
  • Galvanometric Z-stage and motorized XY-stage for multiposition imaging (mosaic, time lapse).
  • White Light Laser (470-670 nm tunable excitation with 1nm interval).
  • Argon Laser  (458, 476, 488, 496, 514 excitation lines).
  • Ultraviolet Laser (405 nm excitation line, AOTF Flexible)
  • STED CW unit (592 depletion laser for super-resolution and 100x objective)
  • Galvanometric Scanner (up to 1400 Hz) and Resonant Scanner (8000 Hz) for fast scanning.
  • High-sensitive Hybrid detectors (x2) and  standard Photomultiplier tubes (PMT) (x2): 4 channel simultaneous acquisition with freely adjustable spectral detection.
  • Environmental control chamber (temperature, CO2, humidity) for in vivo imaging.

Policy of Use and fees of Leica TCS STED CW SP8 microscope.

ApoTome2™

Achucarro's Zeiss ApoTome2

We have a Carl Zeiss ApoTome2 equipment for creating optical sections of fluorescent samples (free of scattered light). The structured illumination of this equipment projects a grid structure into the focal plane of the specimen, and then moves it into three positions using a scanning mechanism.

Main technical specifications

  • Automatic grid selection for optimum contrast with each objective (2.5 – Dry 10 - Dry 20 – Dry 40 - Dry 40 – Oil 65 - Oil)
  • Freely selectable grid, depending on the specimen
  • Combination of multiple contrast techniques: Fluorescence channels (DAPI, GFP, DsRed), brightfield, darkfield, DIC, polarized light.
  • Live Cell Imaging and time lapse (with CO2 and temperature control system)
  • Up to three images per second (depending on the exposure time). Doubling of frame rate with "burst mode" (in Live Cell Imaging)

Ultramicrotome (for electron microscopy)

PT-PC PowerTome Ultramicrotome

Our Imaging Facility is equipped with a PC controlled ultramicrotome by RMC Boeckeler, with an integrated touch screen control monitor, that provides good training capabilities. This equipment can take ultra-thin sections to be then studied in the Electronic Microscopy Laboratory, within the Advanced Research Facilities (SGIker) of the University, in the campus.

Main technical specifications

  • Auto thin sectioning from 1nm and Auto thick sectioning to 15µm
  • Cutting speed range from 0.1 – 100mm/sec
  • Specimen auto feed 200µm
  • High precision manually operated knife stage with 30mm lateral and 12mm forward motion
  • Upper knife stage with 360° rotation and knife clearance angle adjustment from -2° to +15°.
  • Accepts glass knives up to 12mm wide, triangular tungsten carbide knives,...
  • Cutting zone set by its control system that can be adjusted at any time.
  • Numeric & digital displays of number of sections cut, total specimen feed advanced, section thickness, cutting speed and specimen feed remaining
  • Stemi 2000 stereomicroscope with 7.7:1 zoom range, 6.5X to 50X magnification with 10X wide field (23mm) eyepieces