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Confocal Microscopy Unit

The Institute of the Hospital Universitario 12 de Octubre is supported by the Confocal Microscopy Unit of said hospital. This Unit is located in the “Research Center” building of the hospital. The Managers of this Confocal Microscopy Unit are Dr. Fernando Bartolomé and Dr. Gema Ruiz Hurtado.

The responsibilities of the Unit’s Managers are detailed below:

  • Defining the general rules applicable to the use of the Confocal Microscopy to achieve a considerable improvement in its performance and use.
  • Supervising the necessary annual external maintenance activities carried out on the equipment to ensure its correct performance, as well as all communications made with the technical services, when necessary.
  • Counseling the different research groups in the development and commissioning of experiments and protocols for which the equipment must be used.
  • Specific training of new users (initial training).

 

The Confocal Microscopy Unit allows the researchers of the i+12 Institute to implement the following techniques and methodologies:

Confocal Microscopy

    1. In vivo physiological assays.
      1. Quantification of intracellular calcium (Fluo-3AM, Fluo-4AM, MagFluo-4, etc.).
      2. Determination of the plasma (di4ANEPS) and mitochondrial membrane (TMRM, TMRE) potential.
      3. Determinations of oxidative stress (DHE, GSH, Mitosox, Lipid peroxidation, etc.).
      4. FAD/Redox.
      5. Protein colocalization and interaction (FRET).
    2. Immunofluorescence and probe detection (apoptosis and cell proliferation, cellular and subcellular localization, diffusion assays, etc.).
    3. Subcellular localization with fluorescent proteins.
    4. Tridimensional studies (Z-Stack).
    5. Counterstaining: labeling of the Golgi complex (HPA lecithin, protein Grasp68), the endoplasmic reticulum (Ryanodine, Calnexin), the mitochondria (Mitotracker Red/Green), the actin microfilaments (falotoxin) and nuclear labeling (IP, Hoechst, DAPI, etc.).

 

 

To apply these techniques and methodologies, the Confocal Microscopy Unit of the i+12 Institute features the following high-tech equipment:

  • LSM510 META ConfoCor 3 Spectral Confocal Microscope (ZEISS).

 

Gema Ruiz Hurtado. gemaruiz@h12o.es; Telephone No.: 91 390 80 01

Fernando Bartolomé Robledo. fbartolome.imas12@h12o.es; Telephone No.: 91 390 87 65