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The Naples CNR Euro-BioImaging facility

Welcome to the Naples Euro-BioImaging facility located at the IBP in CNR Research area NA-1. The facility is a continuation of the imaging facility developed by Alberto Luini and colleagues at the Mario Negri Sud Institute for Biomedical Research in the 1990s that pioneered the development of the modern Correlative video-Light Electron Microscopy (CLEM).

It is the coordinating facility of the candidate Italian multi-modal multi-sited cellular/ biological Node of Euro-BioImaging, a pan European research infrastructure project (for more information see: http://www.eurobioimaging.eu/).  It is managed by the IBP and operated by the IBP and IMM, institutes of the National Research Council of Italy located at the Castellino campus of the CNR Research area NA-1. The Naples Euro-BioImaging facility (and all other Euro-BioImaging facilities in the participating countries) started their interim operation from May 2016. The Naples facility can be accessed through the Euro-BioImaging portal 

The facility offers service and access to imaging resources in the following areas:

 

Technologies
Light and electron microscopy, correlative microscopy, super-resolution microscopy, high throughput microscopy, Raman microscopy, mass spectrometry-based imaging, microfluidics, microinjection, laser microdissection and advanced image analysis. The main area of expertise remains correlative microscopy with developments aimed at integrating different types of light microscopy with electron or Raman or mass-spectrometry-based imaging. 

The major technologies that are available in the Institute comprise:

Routine light microscopy applications

  • Bright-field and phase-contrast microscopy, for evaluation of general morphology and enzyme-based labelling of cells and tissues;
  • Wide-field fluorescence microscopy, for analysis of fluorescent marker distributions in cells and tissues;
  • Confocal microscopy, to investigate co-localisation of proteins and their three-dimensional (3D) space distribution.

Advanced light microscopy applications

  • Live-cell imaging (time-lapse microscopy), for monitoring intracellular organelles and protein dynamics in time and in 3D space;
  • Fluorescence recovery after photobleaching (FRAP) for quantitative evaluation of protein mobility within organelles, rates of inter-organelle exchange, and protein binding kinetics;
  • Fluorescence resonance energy transfer (FRET), to evaluate the strength and intracellular topology of protein-protein interactions, and to monitor intracellular dynamics of small signalling molecules and protein activities;
  • Cell microinjection, for acute modulation (activation or inhibition) of protein activities.

Routine electron microscopy applications

  • Electron microscopy of thin plastic sections, for analysis of general cell and/or tissue ultrastructure;
  • Whole-mount electron microscopy preparation for investigation of ultrastructural changes in fractionated organelles or viral particles.

Advanced electron microscopy applications

  • Immuno-electron microscopy for ultrastructural evaluation of localisation (and co-localisation) of proteins of interest;
  • Correlative light-electron microscopy (CLEM), for electron microscopy analyses of structures observed in living cells before fixation, and of individual cells with various expression levels genes of interest, or particular intracellular distributions of a gene product.
  • Electron Tomography (ET) for 3D study of complex cellular structures, such as organelles, cytoskeleton, and chromosomes. It recovers the specimen’s 3D structure via computerized tomographic reconstruction from a set of 2D projections obtained with Transmission Electron Microscopy (TEM) at different tilt angles.

Data analysis

  • Development of new image analysis software tools;
  • Morphometric analysis of co-localisation, and intracellular protein distributions and dynamics;
  • Electron microscopy-based morphometric analyses of the shape and size of cellular organelles and extracellular structures, and evaluation of local concentrations of gene products after their expression in cells of interest.


Applications
The main areas of expertise are: cell biology, membrane trafficking, signalling, cancer biology and immunology.
 

Training
The Euro-BioImaging facility at IBP organizes programs aimed at training young researchers in the use of basic as well as advanced microscopic techniques. We organize a yearly course on the theory and practice of microscopy for the prospective regular users of the facility. In addition, requests for special training sessions are also supported by appropriate training programs.
 

We also organize workshops/meetings on imaging and microscopy. The most recent meeting organized by this facility was the EMBO workshop on Cellular imaging of Lipids, 2-6 June 2014, Vico Equense, Italy (http://events.embo.org/14-imaging-lipids/).

Facility