Services

List of Services:


1. Conventional Transmission Microscopy and Epi-fluorescence Microscopy

  • Image Acquisition using transmitted light techniques: bright field, phase contrast, and DIC..

  • Fluorescence Images Acquisition: From fixed specimens with one or several fluorochromes (single or multiple labeling, respectively).

2. Multispectral laser confocal microscopy.

  • Multiple labeling: acquisition of high-resolution, sharpness and contrast images from fixed specimens, presenting multiple fluorescent markers without overlap of signals through tailor-made detection of fluorochromes emission spectra.




  • Co-localization: It refers to the overlap analysis of the distribution of signals in fluorescent markers specimens, and represents the colocalization degree between fluorescent molecules using biparametric histograms (diagram showing different fluorescence intensities of labeled species of interest ), mask (image of colocalized pixels in both fluorescent labelings),and colocalization coefficient (quantification of the degree of colocalized fluorescent signals).


  • Z-Stack: Obtention of optical sectioning in the Z-axis of the specimen, and images overlapping, then producing maximum intensity projections (MIP) of planes that allow visualization of 3D specimen reconstruction.


  • Tile Scan: acquisition of a mosaic image of the specimen by capture of multiple images in the XY axes.

3. Time Lapse Microscopy Imaging.

  • “In vivo” assays: acquisition of imaging sequences of live specimens with over time transmitted light and/or fluorescence techniques in controlled Co2 and temperature conditions. Also, application of multi-positions parameters and/or optical sectioning, then generating XYT or ZYZT images.

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4. Photo-activation.

  • Photo-activation Assays: Capture of images before photoactivation of live specimens that express the molecule of interest fused to a photoactivatable fluorescent protein, selection and activation of a region of interest (ROI) in the specimen with the corresponding laser line, and acquisition of an images’ sequence over time to analyze the dynamic distribution of the fluorescent molecule of interest.

5. FRAP: Fluorescence Recovery After Photobleaching.

  • Fluorescence Recovery After Photobleaching Assays: Obtaining images before photobleaching of live specimens that express the molecule of interest fused to a fluorescent protein, selection and photobleaching of a region of interest (ROI) in the specimen with the corresponding full power laser line, and obtention of an image sequence over time to analyze mobilization of molecules of interest by fluorescence recovery in the ROI.

  • Data quantification and analysis: normalization of results, recovery fluorescence graphics, calculation of the mobile/immobile fraction and half-time fluorescence recovery.

6. FLIP: Fluorescence Loss in Photobleaching.

  • Fluorescence Loss in Photobleaching Assays: capture of images before photobleaching of living specimens that express the molecule of interest fused to a fluorescence protein, selection and continuous photobleaching of a ROI in the specimen with the corresponding full power laser line, obtention of an images’ sequence over time to analyze the dynamic distribution of the molecule of interest by the loss or overall reduction of fluorescent molecules in the specimen.

  • Data quantification and analysis: normalization of results, obtention of graphics about fluorescence loss of the molecule of interest.

7. FRET: Fluorescence Resonance Energy Transfer.

  • Fluorescence Resonance Energy Transfer: analysis of intramolecular or intermolecular interactions, conformational changes or proteolysis process of the molecule of interest fused to a FRET pair.

  • Technical advice in the sample preparation: selection of the most adequate FRET donor/acceptor pairs between fluorescent proteins or secondary antibodies, analysis of the co-localization between donor and acceptors, study of the partial overlap of the emission spectrum of the donor with the excitation spectrum of the acceptor, and measurement of the donor fluorescence lifetime.

  • Selection of the FRET detection method:
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    -FRET Acceptor Photobleaching method: acquisition of images before donor and acceptor photobleaching of the FRET sample, excited with laser lines; selection of a ROI in the acceptor image; continuous acceptor photobleaching and the later capture of images of the donor and acceptor excited with laser lines.

    -Sensitized Emission Method: capture of reference images of donor and acceptor from control samples with the corresponding wavelength laser lines; images acquisition of the donor and acceptor in the FRET sample with donor excitation laser line, and finally, image acquisition of the acceptor alone in the FRET sample with the acceptor excitation laser line.

  • Data quantification and analysis: normalization of results, obtention of graphics of the donor and acceptor fluorescence intensity, pixel by pixel calculation codified in a pseudocolored image relative to the percentage of FRET efficiency.

8. Laser Microdissection and Catapulting.

  • Fixed cells: UV laser catapulting of cells marked with immunohistochemical or immunofluorescent techniques.

  • Tissue Sections: UV laser microdissection and catapulting in ROI from cryo-forzen tissue sections or FFPE, marked with immunohistochemical or immunofluorescent techniques to be used for later extraction assays of DNA (PCR, mutation analysis, SPs…), RNA (qRT-PCR, expression analysis, microarrays…) or protein (immunoblotting, 2D gels, MALDI-TOF…).

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9. Technical Advice on the experiment design.

The Unit provides technical advice and assessment on the design of experiments and the correct preparation of the samples.

10. Technical support on the images processing and analysis.

We provide technical support on the processing and analysis of images by using different software programs such as Zen, AxioVision, PALMRobo, MetaMorph, NIS-Elements, Image J, and Adobe Photoshop, as well as on the correct interpretation and presentation of the data of the images obtained.

11. Supply of consumable material for “in vivo” assays of confocal laser microscopy and laser capture microdissection.