CNR - Institute of Neuroscience CNR
Institute of Neuroscience
 

Project

Optical and electron microscopy laboratory

Biological research involves the elucidation of the structure, localization and dynamics of every molecule involved in the investigated phenomenon. A strong contribution to this knowledge is given by the morphological analysis of organs and tissues and by the studies of the ultrastructure of cells and subcellular organelles, as well as by the analysis of the localization of proteins and nucleic acids in tissues, cells or specific sub-cellular compartments. The laboratory of optical and electron microscopy is dedicated to provide state-of-the-art imaging facilities and technical support to all investigators of the CNR Institute of Neuroscience and interested investigators on the main university campus.

Service facilities

For fluorecence studies the following microscopy systems are available: confocal laser scanning microscope Bio-Rad MRC1024 mounted on an upright microscope, equipped for the observation and acquisition of images in the inteferential contrast mode (DIC); Zeiss LSM 510 Meta mounted on an Axiovert 200 with CO2 and temperature control. A complete set-up for time-lapse in vivo video-microscopy based on Axiovert 200 microscope and CCD camera MicroMAX 512BFT (Princeton Instruments) with CO2 and temperature control. These two last systems have been set up for high resolution 3D fluorescent imaging and dynamic time-lapse of living cells in multiple fluorescence channels. Microinjection is also available.

We routinely employ several commercial softwares for colocalization studies, image analysis and processing (Metamorph, ImageJ) and deconvolution (Huygens SVI) as well as in house developed applications for particle distribution analyses and colocalization.

 

For electron microscopy, there are three ultra-microtomes equipped with cryochambers for the production of thin frozen sections; one transmission and one scanning electron microscopes are operating in the unit. Techniques of conventional transmission and scanning electron microscopy are routinely employed as well as immunocytochemistry. High resolution immunocytochemical techniques (pre-embedding. post-embedding, cryo-immunogold) are applyed to localize proteins at subcellular level.

Our experienced staff can carry out all the steps of sample preparation and analysis for a set fee. Alternatively, our facility is open to users who have had prior training in electron microscopy. Training can be provided by our staff on a one-on-one basis throughout the year.

Current research

 

In collaboration with the laboratories of the Unit, we are particularly involved in the study of the dynamics and structure of cellular organelles. The figures report the salient results obtained in these years. Figure 1 gives an example of a dynamic endoplasmic reticulum restructuration and Figure 2 reports the selective mitochondrial modifications that occur in motoneurons in an animal model of Amiotrophic Lateral Sclerosis.

Publications

  • Bianco F, Perrotta C, Novellino L, Francolini M, Riganti L, Menna E, Saglietti L, Schuchman EH, Furlan R, Clementi E, Matteoli M, Verderio C (2009) Acid sphingomyelinase activity triggers microparticle release from glial cells. EMBO J. 28:1043-54.
  • Massari S, Perego C, Padovano V, D'Amico A, Raimondi A, Francolini M, Pietrini G (2009) LIN7 mediates the recruitment of IRSp53 to tight junctions. Traffic 10:246-57.
  • Ronchi P, Colombo S, Francolini M, Borgese N (2008) Transmembrane domain-dependent partitioning of membrane proteins within the endoplasmic reticulum. J. Cell Biol. 181:105-18.
  • Borgese N, Francolini M, Snapp E (2006) Endoplasmic reticulum architecture: structures in flux. Curr. Opin. Cell Biol. 18:358-64.
  • Ceppi P, Colombo S, Francolini M, Raimondo F, Borgese N, Masserini M (2005) Two tail-anchored protein variants, differing in transmembrane domain length and intracellular sorting, interact differently with lipids. Proc. Natl. Acad. Sci. U.S.A. 102:16269-74.

Grants

Grant della Fondazione Italo Monzino, Milano, per la costituzione del Laboratorio di Microscopia ottica avanzata, 2007

 

PI photo

Maura Francolini

Contact information

email  E-mail

email  +39 02 5031 6976

Participating staff
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