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Electron Microscopy

Electron Microscopy

Unit info

The modern ultrastructural analysis is extremely important for the study of cancer biology. Electron microscopy (EM) has a very high resolution and is able to observe structures not visible under the light microscope. For example, the evaluation of the role of exosomes (or exovesicles) in tumorigenesis, the study of the function of oncogenes and their role in cellular organization and function. Only EM is able to find invisible under the light microscope morphological alterations induced by knock down or knock out of several oncogenes.

At IFOM we have the whole spectrum of methods suitable for the study of above problems. Our unit can perform not only routine transmission and scanning EM, but also all kinds of immune EM, correlative light electron microscopy (CLEM), three-dimensional EM (3DEM), quantitative stereological evaluation of data and so on. For instance, we can label cells using cryo-sections or using pre-embedding methods with nano-gold labelling and subsequent gold enhancements and then estimate the quantitative parameters of the cell examined. We have also developed several methods such as the video CLEM ideal for quickly moving organelles, the discretized rotator for stereological analysis and several new methodology of sample preparation for 3DEM, and so on.

In future we plan to improve methods of CLEM and 3DEM, to develop new quantitative analysis approaches based on absolute stereological estimators. Within this context, several new methods of sample preparation will be proposed in order to meet the specific demands of the researchers in IFOM. The final goal of our effort is to define a new set of guidelines for the use of modern electron microscopy in the specific field of oncology.

Alexandre Mironov

Alexandre Mironov

Alexandre Mironov joined IFOM in 2010. He graduated in Medicine and specialized in Churgery and Pathological Morphology at Ivanovo State Medical Institute (IMSI). After he did his PhD, also at IMSI, in Histology and Embryology, studying mechanisms of kidney preservation. Employed as a senior researcher at Ivanovo State Medical Academy for 5 years, he turned to the study of arteriosclerosis and defended his second thesis on the role of arterial hypertension in tissue biology in 1985. Between 1988 and 1991 Alexandre Miranov assumed positions as professor of Gross Anatomy, Chief of the Electron Microscopic Facility, and President of the Faculty of Therapeutic Science, also at the Academy. In 1994 Mironov moved to Italy and he joined IFOM after 17 years at Consorzio Mario Negri Sud.

 

Lab members

Beznusenko Galina
Micaela Quarto

Photogallery

Electron Microscopy Instruments

Selected Unit Publications

  1. Mironov AA, Beznoussenko GV. 2011.
    Molecular mechanisms responsible for formation of Golgi ribbon.
    Histol Histopathol. 2011 Jan;26(1):117-133.

  2. Kreft ME, Di Giandomenico D, Beznoussenko GV, Resnik N, Mironov AA, Jezernik K. 2010.
    Golgi apparatus fragmentation as a mechanism responsible for uniform delivery of uroplakins to the apical plasma membrane of uroepithelial cells.
    Biol Cell. 102(11):593-607. [PubMed]

  3. Micaroni M, Perinetti G, Berrie CP, Mironov AA. 2010.
    The SPCA1 Ca2+ pump and intracellular membrane trafficking.
    Traffic. 11(10):1315-1333.

  4. Micaroni M, Perinetti G, Di Giandomenico D, Bianchi K, Spaar A, Mironov AA. 2010.
    Synchronous intra-Golgi transport induces the release of Ca2+ from the Golgi apparatus.
    Experimental cell research. 316(13):2071-2086.

  5. Mironov AA [LabMeeting] and Beznoussenko GV. [LabMeeting] 2009.
    Correlative microscopy: a potent tool for the study of rare or unique cellular and tissue events.
    Journal of microscopy 235(3):308-321. [LabMeeting]

  6. San Pietro E, Capestrano M, Polishchuk EV, DiPentima A, Trucco A, Zizza P, Mariggiò S, Pulvirenti T, Sallese M, Tete S, Mironov AA, Leslie CC, Corda D, Luini A, and Polishchuk RS. 2009.
    Group IV phospholipase A(2)alpha controls the formation of inter-cisternal continuities involved in intra-Golgi transport.
    PLoS Biol. 2009 Sep;7(9):e1000194. [PubMed]

  7. Zhang L, Lee SY, Beznoussenko GV, Peters PJ, Yang JS, Gilbert HY, Brass AL, Elledge SJ, Isaacs SN, Moss B, Mironov A, Hsu VW. 2009.
    A role for the host coatomer and KDEL receptor in early vaccinia biogenesis.
    PNAS. 106(1):163-168.

  8. Pulvirenti T, Giannotta M, Capestrano M, Capitani M, Pisanu A, Polishchuk RS, Pietro ES, Beznoussenko GV, Mironov AA, Turacchio G, Hsu VW, Sallese M, and Luini A. 2008.
    A traffic-activated Golgi-based signalling circuit coordinates the secretory pathway.
    Nature cell biology. 10(8):912-922.

  9. Yang J.-S., Gad, H., Lee, K., Mironov A. Zhang, L., Beznoussenko, G.V., Valente, C., Turacchio G., Bonsra, A.N., Du, G., Bourgoin, S., Frohman, M. A., Luini, A., and Hsu V.W. 2008.
    A role for phosphatidic acid in COPI vesicle fission yields insights into Golgi maintenance.
    Nat Cell Biol. 10(10):1146-1153.

  10. Pulvirenti T, Giannotta M, Capestrano M, Capitani M, Pisanu A, Polishchuk RS, San Pietro E, Beznoussenko GV, Mironov AA, Turacchio G, Hsu VW, Sallese M, Luini A. 2008.
    A traffic-activated Golgi-based signalling circuit coordinates the secretory pathway.
    Nat Cell Biol. 10(8):912-922. [biomed experts]

  11. Luini A., Mironov A.A., Polishchuk E.V. and Polishchuk R.S. 2008.
    Morphogenesis of post-Golgi transport carriers.
    Histochem Cell Biol. 129(2):153-161.

  12. Beznoussenko GV, Dolgikh VV, Seliverstova EV, Semenov PB, Tokarev YS, Trucco A, Micaroni M, Di Giandomenico D, Auinger P, Senderskiy IV, Skarlato SO, Snigirevskaya ES, Komissarchik YY, Pavelka M, De Matteis MA, Luini A, Sokolova YY, Mironov AA. 2007.
    Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function.
    J Cell Sci. 120:1288-1298. [PubMed]

  13. Marra P, Salvatore L, Mironov A Jr, Di Campli A, Di Tullio G, Trucco A, Beznoussenko G, Mironov A*, De Matteis MA*. 2007.
    The Biogenesis of the Golgi Ribbon: The Roles of Membrane Input from the ER and of GM130.
    Mol Biol Cell. Feb 21.
    *corresponding authors

  14. Derganc, J., A. A. Mironov, and S. Svetina, 2006.
    Physical factors that affect the number and size of Golgi cisternae.
    Traffic. 7:85-96.

  15. Dolgikh V.V. Semenov P.B., Mironov A.A. Beznoussenko G.V. 2005.
    Immunocytochemical identification of the major exospore protein and three polar-tube proteins of the microsporidia Paranosema grylli.
    Protist. 156:77-87.

  16. Mironov A.A., Beznoussenko G.V., Luini A. and Polishchuk, R.S. 2006.
    Visualizing intracellular events in vivo by combined video fluorescence and 3-D electron microscopy.
    Methods in Enzymology 404:43-57.

  17. Mironov, A.A., Beznoussenko, G.V., Polishchuk, R.S. Trucco, A. 2005.
    Intra-Golgi transport. A way to a new paradigm?
    BBA - Molecular Cell Research, 1744: 340-350.

    18.