IMAGING ACQUISITION AND ANALYSIS

Fluorescence Resonance Energy Transfer (FRET)

Image Acquisition System

Inverted Zeiss Axiovert 200 microscope optimized both for image acquisition with transmitted light and fluorescence and for FRET measurements. The microscope holds standard filters (DAPI/FITC/TRITC), a quick wavelength exchange monochromator as excitation system (OptoScan, Cairn Research), and a high sensitivity and velocity camera (ORCA FLASH 4.0, Hamamatsu).

In order to perform FRET analysis, the microscope is also equipped with the double-emission W-View Gemini system and a CFP/YFP filter set. These features allow two different wavelength images with a single camera. The software of the microscope is in charge of image acquisition and processing (HCImage, Hamamatsu).

Some of the applications of this system are:

  • Transmitted light and fluorescence image acquisition in histological and cytological samples.
  • Real time fluorescence image acquisition in cells through fluorescent probes in order to analyze a wide range of biological parameters (intracellular calcium, pH, oxidative stress, and so on).
  • Receptor-ligand, protein-protein or protein-membrane lipids molecular interaction measurements.
  • Dynamics, conformational changes and protein formation studies.
  • Molecular interaction distances and supramolecular dynamic analysis.
  • Intracellular signalling molecules and surface proteins organization and interaction.
  • Intracellular signalling molecules dynamics (i.e., AMPc/PKA).

This equipment has been acquired thanks to an intervention co-financiated by the European Union through the Programa Operativo del Fondo Europeo de Desarrollo Regional (FEDER) from the Comunitat Valenciana 2014-2020 (PROJECT: IDIFEDER2020/022).

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Miguel Hernández University, Elche
Institute of Research, Development and Innovation in Healthcare Biotechnology in Elche (IDiBE)
Torregaitán Building. Instrumentation Room 1.07 Confocal Microscopy II