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Scientific Programmes

Diego Megías

Scientific Programmes

Biotechnology Programme

Confocal Microscopy Core Unit

Head of Unit:  Diego Megías
Research hightlights

The Confocal Microscopy Core Unit is equipped with 3 laser scanning confocal systems (Leica SP2 and SP5) that incorporate UV and multiphoton excitation, as well as a white light laser and a Hybrid Detector. The Unit is also equipped with 2 wide field systems, namely, a Deltavision 4D deconvolution station and a Leica DMRI6000 system, equipped with microinjection. All the microscopes are automated and equipped with incubators for live-cell imaging.

In addition, the Unit has applied high-throughput (HT) technologies to confocal microscopy using 2 different systems:

  • An Opera (Perkin Elmer) High-Throughput Content Screening (HTS) system that allows running HTS experiments on fixed and live cells in multi-well plates, and enables the monitoring of cell dynamics (translocation, cell division, etc.) by means of fluorescence markers.
  • A Matrix Screening Application integrated into the SP5 confocal systems, allowing high content feeding of the instrument, not only in multi-well plates but also in tissue sections.

These advances increase the level of information obtained from a sample and allow for the automated HTS of cell behaviour in response to di!erent treatments.

Figure 3D-reconstruction of a micro-well – 80 microns high and 80 microns wide – that contains a single living cell on a microarray composed of 4,000 poly-dimethyl-siloxane wells.

During 2012, the Confocal Microscopy Core Unit has continued to establish numerous scientific collaborations with CNIO researchers, covering several aspects of cancer studies such as the Intelligent Screening of single cell behaviour in cellular arrays in vivo and in tissues. In addition, the Unit is developing new methods for automated high resolution acquisition and analysis of cell invasion studies, and is assisting researchers in the measurement of protein activity reporters using Fluorescence Resonance Energy Transfer (FRET) technology. Moreover, the Unit is dedicating a significant e!ort towards the development and implementation of HTS technology at CNIO; for example, during this last year we helped to run screenings aimed at testing compounds that may eventually modify key aspects of tumourigenesis such as mitotic checkpoint regulation, integrity of nucleoli, DNA damage, cell survival and proliferation.