The Advanced Imaging Core Facility (AICF) provides access to a wide collection of frontline light microscopes allowing the users to spam from basic observations with transmitted light to the more advanced fluorescence live imaging techniques. The Facility staff offer specialized technical and scientific support throughout every step of imaging experiments, ranging from experimental design and image acquisitions to image processing and data analysis.
Applications and services
- The applications and services offered by the Facility are as follows:
- Super-resolution microscopy: PALM, dSTORM, SIM and TIRF-SIM
- Confocal microscopy: laser scanning confocal and spinning disc systems
- Fluorescence wide-field microscopy: up-right and inverted microscopes
- Wide-field optical sectioning with structured illumination (Zeiss Apotome 2 module)
- Time-lapse and in-vivo fluorescence imaging (wide-field and confocal) within incubation systems that permit control of environmental conditions over a long period
- Large-area imaging (mosaics, tiles, or panorama modules) on big samples (such as mouse brain sections or histological preparations)
- Other fluorescence microscopy techniques available: FRAP, FRET, smFRET, Single-Particle Tracking, HILO and TIRF
- Transmitted light contrast methods: Bright-Field, Phase Contrast, Differential Interference Contrast and Dark-Field
- Laser capture micro-dissection of tissues at the single-cell level (both on fixed samples and living cells in cultures)
- Histological sample preparations (automatic tissue processor, embedding center, cutting instruments for tissue sectioning and dedicated fluorescence stereomicroscopes)
- Image processing, macro scripting and data analysis using ImageJ/Fiji, SVI Huygens Core and Essential (for deconvolution analysis and distilling PSF), Leica LasAF/LasX, Zeiss ZEN, and Nikon NIS Elements
The Facility staff will be pleased to offer:
- Technical support in experimental design, microscope purchase, and configuration
- Trainings for the autonomous use of the microscopes
- Shadowing for non-autonomous users
- Image acquisition services
- Image processing, macro scripting and data analysis services
Facility Equipment and Software for data analysis
Wide-field systems
- Leica DMi8 fully motorized inverted microscope equipped with a Lumencor Sola white LED and an Andor Zyla 4 monochromatic sCMOS camera.
- Zeiss Axio Imager M2 fully motorized upright microscope, equipped with the X-CITE 120 metal halide fluorescence lamp and with a monochromatic and a color camera (Zeiss Axiocam 305 mono and Zeiss Axiocam 705 color).
- Zeiss Axio Observer Z1 fully motorized inverted microscope equipped with two light sources (Colibri 2 multicolor LEDs and HXP 120 metal halide fluorescence lamp), ApoTome 2 module for structured illumination and Pecon incubator and control modules for live-cell observations.
Confocal systems
- Leica TCS SP8 laser scanning confocal microscope built upon a Leica DMi8 fully motorized inverted microscope, equipped with: Lumencor Sola white LED, multi-lines lasers, one ultra-sensitive HyD detector plus three conventional PMT detectors and a transmitted light detector. Channel separation is achieved via a fully tunable prism-based spectral detector. Leica LAS X software includes modules for FRET, FRAP and Leica’s proprietary Adaptive Deconvolution package (Lightning).
- CREST V3 X-light Spinning Disc system built upon a Nikon Eclipse Ti2 fully motorized inverted microscope, equipped with: multi-line CELESTA lasers, two digital cameras (an Andor Zyla 4.2 monochromatic sCMOS camera and an Andor iXon 888 monochromatic EMCCD camera) and an Okolab incubation system for live-cell observations. Nikon AR software includes Nikon’s proprietary deconvolution module (NIS.ai).
- Nikon AX laser scanning confocal system built upon a Nikon Eclipse Ti2 fully motorized inverted microscope, equipped with: multi-line lasers, Nikon D-LEDI multi-line LEDs and Hamamatsu ORCA Flash 4.0 sCMOS monochromatic sensor camera and two ultra-sensitive GaAsP detectors plus two conventional PMT detectors and a transmitted light detector. In addition to wide-field and confocal acquisitions, this system allows also for 2D and 3D SIM (Structured Illumination Microscopy) and 2D TIRF (Total Internal Reflection Fluorescence) SIM due to the presence of the Nikon N-SIM S module. Nikon AR software includes Nikon’s proprietary deconvolution module (NIS.ai).
Super-resolution systems
- Nikon N-SIM S super-resolution system built upon a Nikon Eclipse Ti2 fully motorized inverted microscope, equipped with: multi-line lasers, Nikon D-LEDI multi-line LEDs and Hamamatsu ORCA Flash 4.0 sCMOS monochromatic sensor camera. In addition to wide-field and SIM acquisitions (2D, 3D SIM and 2D TIRF SIM) this system also allows for confocal acquisitions due to the presence of the Nikon AX module. Nikon AR software includes Nikon’s proprietary deconvolution module (NIS.ai).
- ONI Nanoimager fully motorized inverted system equipped with multi-line lasers and LEDs as light sources and with the Hamamatsu ORCA Flash 4.0 sCMOS monochromatic sensor camera. Available techniques: wide-field, TIRF (Total Internal Reflection Fluorescence), HILO (Highly Inclined and Laminated Optical sheet), dSTORM (Stochastic Optical Reconstruction Microscopy), PALM (Photoactivated Localization Microscopy), Single-particle tracking (SPT), Single-molecule FRET (smFRET), DMD-based SIM and confocal microscopy.
Laser-capture micro-dissection system
- Leica LMD6500 laser micro-dissection system built upon a Leica DM6000 B inverted microscope.
Stereomicroscopes
- Leica MZ 10F, 16F and Leica S9i fully manual fluorescence stereomicroscopes.
Histological instruments
- Leica CM 1850 UV cryostat.
- Thermo Scientific HM525 NX cryostat.
- Leica HistoCore Biocut microtome.
- Leica VT 1200 vibratome.
- Histo-Line HistoPro200 Tissue Processor.
- Histo-Line TEC 2900 Tissue Embedding Center.