The Advanced Imaging and Microscopy Service (SIMA) is a service affiliated with the Institute of Biomedical Technologies (ITB) and the University of La Laguna (ULL). Its objective is to provide ULL research groups, public and private organizations, and local and national companies with the equipment and technical support necessary to perform advanced microscopy experiments (Epifluorescence, Confocal, Phase Contrast and Normarski, FRAT, FRET, etc.), both on fixed samples and live cells. Furthermore, it aims to facilitate the analysis and quantification of microscopy images generated by users.
- Conventional fluorescence microscopy.
- Super-resolution microscopy (STED and dSTORM)
- Detection of immunocytochemical markings using fluorochrome-labeled antibodies in both cell cultures and histological sections.
- Hybridization in situ with fluorescent probes (FISH)
- Obtaining microsamples from histological sections using laser microdissection.
- Cell analysis in vivo and real time using fluorescent markers and/or fusion proteins and derivatives, under controlled temperature and atmosphere conditions.
- Colocalization studies, internalization and intracellular trafficking.
- Physiological analysis of Ca2+ response.
- Study of protein interactions using the FRET technique (Fluorescence Resonance Energy Transfer)
- Study of protein transport using the FRAP technique (Fluorescence Recovery After Photobleaching)
- Determination of fluorescence spectra (Lambda Scan)
- Image processing and quantification with specialized programs: LAS X, Huygens Deconvolution, ImageJ/ Fiji.
SIMA also performs other tasks such as:
- Maintenance and coordination of the use of the various microscopes belonging to the Service.
- Technical support in the use of microscopes and image processing software for untrained personnel.
- Mandatory training for self-employed users.
- Advice and technical support to users.
- Performing experimental procedures.
- Sample preparation.
OLYMPUS FV1000 CONFOCAL MICROSCOPE:
- Olympus IX81 inverted microscope, with motorized XY stage.
- 3 PMTs (photomultiplier detectors)
- 1 transmitted light detector.
- Mercury lamp for fluorescence.
- Incubation chamber with temperature and CO2 control for live cells.
- Objectives: 10X/0.3 NA, 20X/0.75 NA, 40X/1.30 NA and 60X/1.35 NA.
- Fluorescence filters for DAPI, FITC and TRITC.
- Laser lines: 405 nm laser diode, 458/488/514 nm multi-line Argon laser, 543 nm green Helium Neon laser, and 633 nm red Helium Neon laser.
- 4096×4096 scanning system.
- Sweep speed of 4fps at 512×512.
LEICA DM 4000B DIRECT EPIFLUORESCENCE MICROSCOPE:
- 5X/0.15 NA, 10X/0.30 NA, 20X/0.5 NA, 40X/0.75 NA and 100X/1.30 NA objectives for oil immersion.
- Software for image analysis Q-Win and Q-fluoro.
LEICA TCS SP8 CONFOCAL MICROSCOPE:
- DM6000 inverted microscope, with motorized XY stage and galvanometric Z stage.
- 4 detectors: 2 PMT photomultiplier tubes and 2 HyD hybrids (PMT+GaAsP)
- 1 transmitted light detector.
- Metal halide lamp for fluorescence.
- Incubation chamber with temperature and CO2 control for live cells.
- Objectives: 10X/0.40 NA, 20X/0.70 NA, 40X/0.85 NA, 40X/1.30 NA, and 63X/1.40 NA.
- Fluorescence filters for DAPI, FITC and TRITC.
- Laser lines: 405 nm laser diode, 458/488/514 nm multi-line Argon laser, 561 nm laser diode, and 633 nm red Helium Neon laser.
- 8192×8192 high-resolution scanning system; (x) 4fps scanning speed at 512×512.
- LAS AF software with modules for FRET and FRAP colocation.
STIMULATED EMISSION DEPLETION SUPER-RESOLUTION MICROSCOPY SYSTEM (STED) COUPLED TO THE TCS SP8 MICROSCOPE:
- 2 high-power STED lasers (>1.5 W) with wavelengths of 592 nm and 660 nm, for depletion.
- Optical coupling using ultra-precise optical fiber with "Votex phase filter" and optical polarization for resolution improvement, zero PSF optimization.
- Rack integrating STED laser, electronic control and optical modulation (AOTF)
- Integration into the LAS X confocal software.
- Implementation of deconvolution software with algorithms specifically dedicated to STED images.
- Software package for deconvolution, visualization and analysis of STED and confocal images.
- Target optimized for STED applications, PLAN APO 100x/1.40 STED White oil.
- STED 3D Module.
- Active anti-vibration table.
- Active optical table.
TOTAL INTERNAL REFLECTION MICROSCOPY (TIRF) UNIT COUPLED TO ELECTROPHYSIOLOGY AND SPECTROSCOPY.
- Complete electrophysiology system (patch-clamp. Axon/Molecular Devices) coupled to a TiU microscope equipped with a TIRF module, DMD system, SpectraX LED illumination (Lumencor) with filter sets for all wavelengths available in motorized filter turrets.
- Imaging system composed of an Andor IXON EMCCD camera, coupled to an ACTON spectrograph with diffraction gratings and mirror.
- NiDAQ interface, MicroManager software and NIS-Elements (NIKON)
- System coupled to two possible perfusion devices, one ultra-fast (multichannel systems) and the other conventional (ALA systems)
LEICA AS LMD LASER MICRODISECTION SYSTEM.
- Complete standard equipment for performing laser capture microdissection to isolate regions or cells of interest.
| SERVICES OFFERED | PRICES (€) | ||
| ITB | ULL | GENERAL | |
| Observation, analysis and image | |||
| Leica SP8 | 15 | 20 | 25 |
| Olympus FV 1000 | 10 | 15 | 20 |
| Conventional/Fluorescence Microscopy | 7 | 12 | 15 |
| Data processing | – | – | – |
| Image processing and analysis/hour | 12 | 20 | 35 |
| Other features | – | – | – |
| Technical support/hour | 11 | 16 | 22 |