National Facility for Light Imaging

Wide-field microscopy is a widely used technique which allows for imaging the sample using many different contrast methods, such as fluorescence, bright- and dark-field, differential interferometric contrast (DIC) or phase contrast.

Available wide-field microscopes:

• Leica Thunder Imager Live Cell
  Motorized, inverted wide-field microscope with digital clearing. Epifluorecence illuminator with 8 single LEDs (395 nm, 438 nm, 475 nm, 551 nm, 555 nm, 575 nm, 635 nm and 730 nm) and sCMOS camera.
• Zeiss Axioscope5
  Manual, upright wide-field microscope with LED epifluorescence illuminator and monochrome sCMOS camera, condenser for phase and darkfield imaging.
• Zeiss Axiozoom V16 with apotome
  Binocular microscope with motorized stage and focus for large FOV acquisition, back-illuminated sCMOS camera and Apotome module.

Confocal microscopy allows for rejecting out-of-focus light, improving optical sectioning and enabling three-dimensional reconstruction.

Available confocal microscopes:

• Zeiss LSM980
  Point-scanning confocal microscope with Airyscan2 detector, 5 laser lines (405 nm, 488 nm, 561 nm, 594 nm, and 639 nm), sample finder, epifluorescence excitation and sCMOS camera.
• Zeiss LSM980-NLO
  Point-scanning confocal microscope with multiphoton excitation and Airyscan2 detector, 5 laser lines (405 nm, 488 nm, 561 nm, 594 nm, and 639 nm), multiphoton excitation with a tunable Ti:Sa laser (700 nm – 1300 nm) and a fixed (1040 nm) laser line, 6 non-descanned detectors (4 PMT and 2 BiG), epifluorescence excitation and sCMOS camera.
• Nikon Ti2 with CREST v3 spinning disk scan head
  Spinning disk confocal microscope with 7 excitation laser lines (405 nm,  446 nm, 477 nm, 520 nm, 547 nm, 638 nm, and 749 nm), 4 high-power laser lines for FRAP and TIRF (405 nm, 488 nm, 520 nm, and 640 nm), 4 high-speed, large FOV back-illuminated sCMOS cameras.
• Leica Stellaris 8 for live-cell imaging
  Point-scanning confocal microscope with white light laser (up to 8 simultaneous laser lines between 440 nm and 790nm), 405 nm laser, AOBS tunable dichroic, dual scanner (conventional and resonant) and integrated FLIM module.

Super-resolution microscopy allows for reaching a resolution beyond the diffraction limit.

Available super-resolution microscopes:

• Zeiss Elyra7
  Lattice structured illumination and localization microscope with 4 laser lines (405 nm, 488 nm, 561 nm, and 638 nm), dual sCMOS cameras, TIRF, PALM, and STORM modules
• Abberrior FacilityLine
  STED microscope with 4 laser sources (405 nm, 485 nm, 561 nm, and 640 nm), depletion laser (755 nm), adaptive optics system, MATRIX detectors and FLIM module
• Leica Stellaris 8 for super-resolution imaging
  Confocal microscope with STED module and tandem scanners (conventional and resonant), white laser (up to 8 simultaneous laser lines between 440 nm and 790nm), 405 nm laser, AOBS tunable dichroic and integrated FLIM module.

Lightsheet microscopy minimizes phototoxicity and photobleaching by selectively exposing only the area of the sample being imaged.

Available lightsheet microscopes:

• Zeiss Lattice Lightsheet 7
  Lattice lightsheet with 3 lasers (488 nm, 561 nm, and 640 nm) and dual cameras.

Slide scanners allows for automated imaging of many slides

• Zeiss Axio Scan.Z1 automated slide scanner
  Automated slide scanner with color and monochrome cameras, LED epifluorescence illuminator with 7 LEDs (385 nm, 430 nm, 475 nm, 555 nm, 590 nm, 630 nm, and 735 nm), 6 objectives (2.5x/0.12, 5x/025, 10x/0.45, 20x/0.45, 20x/0.8, and 40x/0.95).

Full-service sorting of rare populations from heterogeneous samples, cell cloning (single cell deposition into multi-well plates), particle enrichment, and high purity bulk sorts.

• High-recovery and indexed single-cell sorting for sequencing.
• Cell sorting of many cell types including immune cell and hematopoietic stem cell subsets, mesenchymal stem cells, viable cytokine producing cells and general cell sorting approaches for cell lines and transfected cells.

Sorter technical features:

• BD FACSDiscover S8
  This instrument is the latest technological advance, representing a breakthrough solution to combine capabilities of cell separation with multiparametric spectral parameter-based analysis and imaging.  Image based cell selection combined with fluorescently labelled tags for determination of tag local. Sorting of up to 6 populations simultaneously with recovery in numerous devices including 384 well microplates. The sorter is equipped with 5 lasers (349 nm, 405 nm, 488 nm, 561 nm, and 637 nm).
• MoFlo Astrios EQ
  The sorter features dual forward scatter for enhanced detection of small particles such as MV’s and capable of very high-speed bulk sorting. Sorting of up to 6 populations simultaneously with recovery in numerous devices including 1536 well microplates. The sorter is equipped with 6 lasers (355 nm, 405 nm, 488 nm, 561 nm, 592 nm, and 640 nm).

Analysis of cell populations in suspension using conventional, spectral and imaging analyzers.

Analyzer technical features:

• Cytek Aurora Spectral Analyzer
  The analyzer is designed for high-complexity applications, enabling the use of a wide array of new fluorochrome combinations and capable of resolving challenging dye combinations as well as extracting autofluorescence to enhance resolution of dim markers.  The analyzer is equipped with 5 lasers (355 nm, 405 nm, 488 nm, 561 nm, and 640 nm), spectral detection of 64 channels and is equipped with automated sample loader.
• ImageStream Mark II Imaging Flow Cytometer
  This instrument combines the speed, sensitivity, and phenotyping capabilities of flow cytometry with the detailed imagery and functional insights of microscopy. The analyzer is equipped with 6 lasers (375 nm, 405 nm, 488 nm, 561 nm, 592 nm, and 642 nm) and a 785 nm side scatter laser, a 12-channel spectral detector, and multi-channel brightfield.
• CytoFLEX LX Flow Cytometer
  High speed and multi-parametric analysis for immunophenotyping and micro-vesicle analysis with 6 lasers and 21 fluorescence APD detectors. Laser wavelengths include 355, 405, 488, 561, 640, 808 nm. Sample acquisition may be performed in single tubes or in 96-well plates (flat, V or U bottom). Deep-Well plates may also be used for samples requiring lager volumes. In addition to the standard Side Scatter parameter on the 488 nm laser line, an added side-scatter parameter is configured for the 405 nm laser line.

Autonomous Flow Cytometry Analysis (after completion of a training session) or Operator Assisted Flow Cytometry Analysis.

MEA arrays measure the electrical activity of large cellular networks in real time without labelling.

• 3BRAIN AG BioCAM DupleX
  MEA array system for in vitro functional imaging, compatible with MEA chips with 4096 electrodes. The system works with adherent cells, tissue and organoids slices and allows for recording extracellular voltage oscillations.

Using a fluorescent ion sensor, it is possible to analyze the electrical activity of a cell by measuring the intracellular ion oscillations using a conventional microscope. This service will be performed using microscopes of the IU1 Imaging.