Scientific-Technical Facilities
Advanced Optical Microscopy
Facility Description
Advanced Light Microscopy Facility provides its services to both members of the institute itself and external users, whether they belong to public organizations or private entities. This service includes the start-up of the equipment as well as technical assistance.
Fluorescence or confocal microscopy can be used to analyse very diverse types of samples. In our facility, it is generally used to analyse in vivo cultures of cyanobacteria or microalgae, vegetal tissues (leaves, roots and stems) or entire seedlings as well as cut sections of vegetal and animal tissues, and sometimes metal surfaces.
The most common types of studies in this regard are as follows:
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- In vivo protein subcellular localisation detected via fusion with fluorescent proteins (e.g., GFP, YFP, CFP).
- Cell expression studies employing promoter fusion with reporter genes of fluorescent proteins.
- Serial sections for 2D and 3D pseudo-reconstruction.
- BIFC studies.
- Detection of inmunocytochemical markers.
- Colocalisation analysis.
- Time-lapse experiments.
- FRET and FRAP analysis.
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Equipment
Advanced Light Microscopy Facility has a Leica DM6000B fluorescence microscope and an Olympus FLOUVIEW FV3000 Laser Scanning Confocal Microscope (LSCM) with the following specifications:
- Leica DM6000B Fluorescence Microscope:
- Software: Leica LAS AX with deconvolution module.
- Ilumination: Halogen lamp 12V-100W and Mercury Short Arc LampHPX®120 W.
- Fluorescence filter systems: A (DAPI-LP), +A4 (DAPI), +L5 (FITC), +TX2 (TEXA-RED), Y7 (CY7), CFP, YFP.
- Objectives Lenses: HCX PL FLUOTAR 10x/0.30, HCX PL S-APO 40x/0.75, HCX PL APO 63x/1.30 Oil, HCX PL APO 100x/1.40-0.70 Oil.
- Digital Camera: Hamamatsu ORCA-ER (Mod. C4742-80-12AG).
- Olympus FLOUVIEW FV3000 Laser Scanning Confocal Microscope (LSCM):
- Microscope: Olympus IX83 fluorescence microscope with motorized stage in XYZ (Macro to Micro image).
- Software: FV31S-SW acquisition software with Super-Resolution module (TruSigthTM) reaching lateral resolution up to 120 nm and cellSens analysis software with FRET and FRAP analysis and 3D deconvolution modules.
- Epifluorescence: pE-300white ilumination. Fluorescence filter systems: U-FUW (DAPI), U-FBN (FITC) y U-FGWA (Rhodamine).
- Diode lasers: 405 nm (50mW), 445 nm (75 mW), 488 nm (20mW), 514 nm (40 mW), 561 nm (20 mW), 640 nm (40 mW).
- Hybrid scanner unit: galvanometer scanner (16 fps @512×512) and resonant scanner (438 fps 512×32 y 30 fps @512×512).
- Detectors: 4 fluorescence PMT (2 GaAsP-PMT y 2 multi-alkali-PMT) y 1 transmitted light PMT.
- Objective Lenses: PLANPON 1.25X/0.04 DRY, X-Line 4x/0.1 3, X-Line 10x/0.4 DRY.A-Line 30x/1.05 Silicone, X-line 40x/1.4 Oil, A-Line 60x/1.5 Oil.
Operating Standards
Reservations for use of the Advanced Light Microscopy Facility must be booked at least 7 days in advance by calling 954489611 or by sending an e-mail to .
Both an application form and an authorization form must be completed and delivered to the personnel in charge of the service being requested.
The services provided and the time (hours) they are in use are billed according to the attached rates.
Personnel assign requested hours according to equipment availability and user waiting lists.
If you require more hours than you originally booked, a new allocation must be solicited.
Cancellations of booked services must be made 48 hours in advance.
The user is responsible for sample preparations. Samples should be prepared and mounted in advance. If it is necessary to mount a sample immediately before its analysis, the user must supply all necessary materials.
Technical assistance is provided in order to analyse the specimen under the best possible conditions.
Before using the confocal microscope, it is recommended that the user analyse specimens in his/her laboratory in order to estimate the level of experimental success that can be achieved.
The results of analyses performed on IBVF equipment are provided in the way agreed upon with the technician.
The results are also temporarily stored on the hard-disks of the equipment and are systematically deleted on a weekly basis.
Rates
|
Marginal Cost |
IIQ-ICMS |
CSIC-US |
AGE-OPIs-UNIV. |
COMPANIES |
Mic. Olympus FV3000 Confocal (with technician) |
17.05 |
18.52 |
20 |
97.13 |
106.38 |
Mic. Fluorescence (with technician) |
0.64 |
0.69 |
0.75 |
47.62 |
79.37 |
Mic. Fluorescence (without technician) |
0.64 |
0.69 |
0.75 |
0.94 |
1.57 |
Prices in €/hour.
Prices valid until march 2025
Request for Facility