Open Lab of Advance Microscopy - OLMA@MBC

OLMA@MBC is an open access facility of the Molecular Biotechnology Center (MBC) of the University of Turin. The facility allows access to state-of-the-art equipment including  widefield and confocal microscopes for the structural and molecular analysis of live and fixed cells or tissues.

OLMA@MBC provides support and user training for these systems.

The microscopes are available to both members of the University of Turin and external users.

 

Services provided:

  • Experimental planning of light microscopy experiments.
  • Training of new users.
  • Technical assistance.
  • Multicolor images acquisition and 3D rendering on confocal microscope.
  • Support for image processing, rendering and analysis.
  • Support for specific imaging techniques.

The facility is open to scientific collaborations on specific projects.

 

Applications:

  • High sensitivity laser scanner confocal microscopy.
  • Multi-photon laser scanner microscopy.
  • Fast sequential acquisition of fluorophores with minimal bleed-through and cross-talk.
  • Environmental control (CO2 and temperature) for in-vivo imaging.
  • 3D z-stacks.
  • Mosaic reconstruction of large samples.
  • dynamic imaging of steady state processes with Photoactivation/ Photobleaching (FRAP, paGFP).
  • sensitised emission FRET assay.
  • quantitative multi-content image acquisition.
Equipment
Leica SP5 inverted confocal microscope with multiphoton laser
  • 7 excitation laser lines (405, 458, 477, 488, 514, 561, 633).
  • 2-photon Tunable Chameleon Ultra 2 laser (680-1080 nm).
  • conventional and resonant (8000 Hz) scanners.
  • 3 PMTs fluorescence detectors and 2 high sensitivity hybrid GaAsP detectors (HyDs).
  • Objectives: 20x / 0,5 dry, 20x / 0,75 imm., 40x / 1,25 oil, 63x / 1,4 oil, 63x / 1,3 glyc.
  • Environmental chamber for live cells.
  • Motorized xy-stage for imaging multiple areas per dish.
  • Leica LAS AF matrix for multiple acquisitions/ multicontent imaging.

 

Leica SP8 inverted confocal microscope
  • 7 excitation laser lines: 458, 477, 488, 496, 514, 561, 633 nm.
  • 2 PMTs fluorescence detectors and 2 high sensitivity hybrid GaAsP detectors (HyDs).
  • AOBS based system.
  • Objectives: 20x / 0,5 dry, 40x / 1,25 oil, 63x / 1,4 oil.
  • Auto Focus Control.
  • LAS_X Leica confocal software with 3D/4D reconstruction module.
Widefield microscopes

Zeiss Axio Observer:

  • Inverted microscope
  • DIC (Differential interference contrast)
  • Filter sets for wide variety of fluorophores
  • 5x dry to 100x oil objectives
  • Apotome
  • Slide and multi-well plate holder

Nikon Eclipse 80i-ViCO:

  • Upright microscope
  • DIC (Differential interference contrast)
  • Filter sets for wide variety of fluorophores
  • Objectives: 20x, 40x (oil), 60x (oil)
  • ViCO

Zeiss Axio Observer- Live cell station:

  • Inverted microscope
  • Environmental chamber for live cells
  • Phase contrast
  • Filter sets for wide variety of fluorophores
  • 5x dry to 100x oil objectives
  • Slide and multi-well plate holder
Publications

Below you can find some of the publications that have included data obtained within the OLMA@MBC.

 

2017

Avalle L, Incarnato D, Savino A, Gai M, Marino F, Pensa S, Barbieri I, Stadler MB, Provero P, Oliviero S, Poli V.

MicroRNAs-143 and -145 induce epithelial to mesenchymal transition and modulate the expression of junction proteins.

Cell Death Differ. 2017 Jun 23. doi: 10.1038/cdd.2017.103

 

Bianchi FT, Tocco C, Pallavicini G, Liu Y, Vernì F, Merigliano C, Bonaccorsi S, El-Assawy N, Priano L, Gai M, Berto GE, Chiotto AM, Sgrò F, Caramello A, Tasca L, Ala U, Neri F, Oliviero S, Mauro A, Geley S, Gatti M, Di Cunto F.

Citron Kinase Deficiency Leads to Chromosomal Instability and TP53-Sensitive Microcephaly.

Cell Rep. 2017 Feb 14;18(7):1674-1686. doi: 10.1016/j.celrep.2017.01.054.

 

Sciortino M, Camacho-Leal MDP, Orso F, Grassi E, Costamagna A, Provero P, Tam W, Turco E, Defilippi P, Taverna D, Cabodi S.

Dysregulation of Blimp1 transcriptional repressor unleashes p130Cas/ErbB2 breast cancer invasion.

Sci Rep. 2017 Apr 25;7(1):1145. doi: 10.1038/s41598-017-01332-z.

 

Panieri E, Millia C, Santoro MM.

Real-time quantification of subcellular H2O2 and glutathione redox potential in living cardiovascular tissues.

Free Radic Biol Med. 2017 Aug;109:189-200. doi: 10.1016/j.freeradbiomed.2017.02.022.

 

Gays D, Hess C, Camporeale A, Ala U, Provero P, Mosimann C, Santoro MM.

An exclusive cellular and molecular network governs intestinal smooth muscle cell differentiation in vertebrates.

Development. 2017 Feb 1;144(3):464-478. doi: 10.1242/dev.133926.

 

Audrito V, Managò A, La Vecchia S, Zamporlini F, Vitale N, Baroni G, Cignetto S, Serra S, Bologna C, Stingi A, Arruga F, Vaisitti T, Massi D, Mandalà M, Raffaelli N, Deaglio S.

The enzyme nicotinamide phosphoribosyltransferase (NAMPT) is up-regulated by BRAF-inhibitor-resistant melanoma cells, becoming an actionable therapeutic target.

JNCI. 2017 (in press).

 

Vaisitti T, Gaudino F, Ouk S, Moscvin M, Vitale N, Serra S, Arruga F, Zakrzewski JL, Liou HC, Allan JN, Furman RR, Deaglio S.

Targeting metabolism and survival in chronic lymphocytic leukemia and Richter syndrome cells by a novel NF-kB inhibitor.

Haematologica. 2017 Aug 31. doi: 10.3324/haematol.2017.173419.

 

Audrito V, Serra S, Stingi A, Orso F, Gaudino F, Bologna C, Neri F, Garaffo G, Nassini R, Baroni G, Rulli E, Massi D, Oliviero S, Piva R, Taverna D, Mandalà M, Deaglio S.

PD-L1 up-regulation in melanoma increases disease aggressiveness and is mediated through miR-17-5p.

Oncotarget. 2017 Feb 28;8(9):15894-15911. doi: 10.18632/oncotarget.15213.

 

Griggio V, Mandili G, Vitale C, Capello M, Macor P, Serra S, Castella B, Peola S, Foglietta M, Drandi D, Omedé P, Sblattero D, Cappello P, Chiarle R, Deaglio S, Boccadoro M, Novelli F, Massaia M, Coscia M.

Humoral immune responses toward tumor-derived antigens in previously untreated patients with chronic lymphocytic leukemia.

Oncotarget. 2017 Jan 10;8(2):3274-3288. doi: 10.18632/oncotarget.13712.

 

2016

Gai M, Bianchi FT, Vagnoni C, Vernì F, Bonaccorsi S, Pasquero S, Berto GE, Sgrò F, Chiotto AM, Annaratone L, Sapino A, Bergo A, Landsberger N, Bond J, Huttner WB, Di Cunto F.

ASPM and CITK regulate spindle orientation by affecting the dynamics of astral microtubules.

EMBO Rep. 2016 Oct;17(10):1396-1409.

 

Sgrò F, Bianchi FT, Falcone M, Pallavicini G, Gai M, Chiotto AM, Berto GE, Turco E, Chang YJ, Huttner WB, Di Cunto F.

Tissue-specific control of midbody microtubule stability by Citron kinase through modulation of TUBB3 phosphorylation.

Cell Death Differ. 2016 May;23(5):801-13. doi: 10.1038/cdd.2015.142

 

Torchiaro E, Lorenzato A, Olivero M, Valdembri D, Gagliardi PA, Gai M, Erriquez J, Serini G, Di Renzo MF.

Peritoneal and hematogenous metastases of ovarian cancer cells are both controlled by the p90RSK through a self-reinforcing cell autonomous mechanism.

Oncotarget. 2016 Jan 5;7(1):712-28. doi: 10.18632/oncotarget.6412

 

Franco I, Margaria JP, De Santis MC, Ranghino A, Monteyne D, Chiaravalli M, Pema M, Campa CC, Ratto E, Gulluni F, Perez-Morga D, Somlo S, Merlo GR, Boletta A, Hirsch E.

Phosphoinositide 3-Kinase-C2α Regulates Polycystin-2 Ciliary Entry and Protects against Kidney Cyst Formation.

J Am Soc Nephrol. 2016 Apr;27(4):1135-44. doi: 10.1681/ASN.2014100967.

 

Zamboni V, Armentano M, Sarò G, Ciraolo E, Ghigo A, Germena G, Umbach A, Valnegri P, Passafaro M, Carabelli V, Gavello D, Bianchi V, D'Adamo P, de Curtis I, El-Assawi N, Mauro A, Priano L, Ferri N, Hirsch E, Merlo GR.

Disruption of ArhGAP15 results in hyperactive Rac1, affects the architecture and function of hippocampal inhibitory neurons and causes cognitive deficits.

Sci Rep. 2016 Oct 7;6:34877. doi: 10.1038/srep34877.

 

Conte D, Garaffo G, Lo Iacono N, Mantero S, Piccolo S, Cordenonsi M, Perez-Morga D, Orecchia V, Poli V, Merlo GR.

The apical ectodermal ridge of the mouse model of ectrodactyly Dlx5;Dlx6-/- shows altered stratification and cell polarity, which are restored by exogenous Wnt5a ligand.

Hum Mol Genet. 2016 Feb 15;25(4):740-54. doi: 10.1093/hmg/ddv514.

 

Orso F, Quirico L, Virga F, Penna E, Dettori D, Cimino D, Coppo R, Grassi E, Elia AR, Brusa D, Deaglio S, Brizzi MF, Stadler MB, Provero P, Caselle M, Taverna D.

miR-214 and miR-148b Targeting Inhibits Dissemination of Melanoma and Breast Cancer.

Cancer Res. 2016 Sep 1;76(17):5151-62. doi: 10.1158/0008-5472.CAN-15-1322.

 

Serra S, Vaisitti T, Audrito V, Bologna C, Buonincontri R, Chen SS, Arruga F, Brusa D, Coscia M, Jaksic O, Inghirami G, Rossi D, Furman RR, Robson SC, Gaidano G, Chiorazzi N, Deaglio S.

Adenosine signaling mediates hypoxic responses in the chronic lymphocytic leukemia microenvironment

Blood Advances. 2016 1:47-61; doi: https://doi.org/10.1182/bloodadvances.

 

Fiorcari S, Maffei R, Audrito V, Martinelli S, Ten Hacken E, Zucchini P, Grisendi G, Potenza L, Luppi M, Burger JA, Deaglio S, Marasca R.

Ibrutinib modifies the function of monocyte/macrophage population in chronic lymphocytic leukemia.

Oncotarget. 2016 Oct 4;7(40):65968-65981. doi: 10.18632/oncotarget.11782.

 

Bologna C, Buonincontri R, Serra S, Vaisitti T, Audrito V, Brusa D, Pagnani A, Coscia M, D'Arena G, Mereu E, Piva R, Furman RR, Rossi D, Gaidano G, Terhorst C, Deaglio S.

SLAMF1 regulation of chemotaxis and autophagy determines CLL patient response.

J Clin Invest. 2016 Jan;126(1):181-94. doi: 10.1172/JCI83013.

 

Pagoto A, Stefania R, Garello F, Arena F, Digilio G, Aime S, Terreno E.

Paramagnetic Phospholipid-Based Micelles Targeting VCAM-1 Receptors for MRI Visualization of Inflammation.

Bioconjug Chem. 2016;27(8):1921-30. doi: 10.1021/acs.bioconjchem.6b00308.

 

Hu H,  Arena F,   Gianolio E,  Boffa C, Di Gregorio E,  Stefania R,  Orio L, Baroni S, Aime S.
Mesoporous silica nanoparticles functionalized with fluorescent and MRI reporters for the visualization of murine tumors overexpressing αvβ3 receptors.
Nanoscale. 2016;8(13):7094-104. doi: 10.1039/c5nr08878j.

Accardo A, Arena F, Gianolio E, Marasco D, Ringhieri P, Boffa C, Bardini P, Aime S, Morelli G.
Diolein Based Nanostructures as Targeted Theranostics.
J Biomed Nanotechnol. 2016;12(5):1076-88.

 

2015

Bergo A, Strollo M, Gai M, Barbiero I, Stefanelli G, Sertic S, Cobolli Gigli C, Di Cunto F, Kilstrup-Nielsen C, Landsberger N.

Methyl-CpG binding protein 2 (MeCP2) localizes at the centrosome and is required for proper mitotic spindle organization.

J Biol Chem. 2015 Feb 6;290(6):3223-37. doi: 10.1074/jbc.M114.608125.

 

Ullio C, Brunk UT, Urani C, Melchioretto P, Bonelli G, Baccino FM, Autelli R.

Autophagy of metallothioneins prevents TNF-induced oxidative stress and toxicity in hepatoma cells.

Autophagy. 2015;11(12):2184-98. doi: 10.1080/15548627.2015.1106662

 

Garaffo G, Conte D, Provero P, Tomaiuolo D, Luo Z, Pinciroli P, Peano C, D'Atri I, Gitton Y, Etzion T, Gothilf Y, Gays D, Santoro MM, Merlo GR.

The Dlx5 and Foxg1 transcription factors, linked via miRNA-9 and -200, are required for the development of the olfactory and GnRH system.

Mol Cell Neurosci. 2015 Sep;68:103-19. doi: 10.1016/j.mcn.2015.04.007.

 

Fiorcari S, Martinelli S, Bulgarelli J, Audrito V, Zucchini P, Colaci E, Potenza L, Narni F, Luppi M, Deaglio S, Marasca R, Maffei R.

Lenalidomide interferes with tumor-promoting properties of nurse-like cells in chronic lymphocytic leukemia.

Haematologica. 2015 Feb;100(2):253-62. doi: 10.3324/haematol.2014.113217.

 

Audrito V, Serra S, Brusa D, Mazzola F, Arruga F, Vaisitti T, Coscia M, Maffei R, Rossi D, Wang T, Inghirami G, Rizzi M, Gaidano G, Garcia JG, Wolberger C, Raffaelli N, Deaglio S.

Extracellular nicotinamide phosphoribosyltransferase (NAMPT) promotes M2 macrophage polarization in chronic lymphocytic leukemia.

Blood. 2015 Jan 1;125(1):111-23. doi: 10.1182/blood-2014-07-589069.

 

Piva R, Deaglio S, Famà R, Buonincontri R, Scarfò I, Bruscaggin A, Mereu E, Serra S, Spina V, Brusa D, Garaffo G, Monti S, Dal Bo M, Marasca R, Arcaini L, Neri A, Gattei V, Paulli M, Tiacci E, Bertoni F, Pileri SA, Foà R, Inghirami G, Gaidano G, Rossi D.

The Krüppel-like factor 2 transcription factor gene is recurrently mutated in splenic marginal zone lymphoma.

Leukemia. 2015 Feb;29(2):503-7. doi: 10.1038/leu.2014.294.

 

Vaisitti T, Audrito V, Serra S, Buonincontri R, Sociali G, Mannino E, Pagnani A, Zucchetto A, Tissino E, Vitale C, Coscia M, Usai C, Pepper C, Gattei V, Bruzzone S, Deaglio S.

The enzymatic activities of CD38 enhance CLL growth and trafficking: implications for therapeutic targeting.

Leukemia. 2015 Feb;29(2):356-68. doi: 10.1038/leu.2014.207.

 

Rizzitelli S, Giustetto P, Cutrin JC, Delli Castelli D, Boffa C, Ruzza M, Menchise V, Molinari F, Aime S, Terreno E.

Sonosensitive theranostic liposomes for preclinical in vivo MRI-guided visualization of doxorubicin release stimulated by pulsed low intensity non-focused ultrasound.

J Control Release. 2015;202:21-30. doi: 10.1016/j.jconrel.2015.01.028.

 

Chirizzi C, Dastrù W, Delli Castelli D, Menchise V, Aime S, Terreno E.

Glucan Particles Loaded with Fluorinated Emulsions: a Sensitivity Improvement for the Visualization of Phagocytic Cells by 19F-MRI.

Current Molecular Imaging. 2015; 4(1):29-34. doi:10.2174/2211555204666150702160805

 

2014

Berto GE, Iobbi C, Camera P, Scarpa E, Iampietro C, Bianchi F, Gai M, Sgrò F, Cristofani F, Gärtner A, Dotti CG, Di Cunto F.

The DCR protein TTC3 affects differentiation and Golgi compactness in neurons through specific actin-regulating pathways.

PLoS One. 2014 Apr 2;9(4):e93721. doi: 10.1371/journal.pone.0093721.

 

Pavan S, Musiani D, Torchiaro E, Migliardi G, Gai M, Di Cunto F, Erriquez J, Olivero M, Di Renzo MF.

HSP27 is required for invasion and metastasis triggered by hepatocyte growth factor.

Int J Cancer. 2014 Mar 15;134(6):1289-99. doi: 10.1002/ijc.28464.

 

Repetto D, Camera P, Melani R, Morello N, Russo I, Calcagno E, Tomasoni R, Bianchi F, Berto G, Giustetto M, Berardi N, Pizzorusso T, Matteoli M, Di Stefano P, Missler M, Turco E, Di Cunto F, Defilippi P.

p140Cap regulates memory and synaptic plasticity through Src-mediated and citron-N-mediated actin reorganization.

J Neurosci. 2014 Jan 22;34(4):1542-53. doi: 10.1523/JNEUROSCI.2341-13.2014

 

Saoncella S, Tassone B, Deklic E, Avolio F, Jon C, Tornillo G, De Luca E, Di Iorio E, Piva R, Cabodi S, Turco E, Pandolfi PP, Calautti E.

Nuclear Akt2 opposes limbal keratinocyte stem cell self-renewal by repressing a FOXO-mTORC1 signaling pathway.

Stem Cells. 2014 Mar;32(3):754-69. doi: 10.1002/stem.1565.

 

Seano G, Chiaverina G, Gagliardi PA, di Blasio L, Puliafito A, Bouvard C, Sessa R, Tarone G, Sorokin L, Helley D, Jain RK, Serini G, Bussolino F, Primo L.

Endothelial podosome rosettes regulate vascular branching in tumour angiogenesis.

Nat Cell Biol. 2014 Oct;16(10):931-41, 1-8. doi: 10.1038/ncb3036.

 

Massi D, Brusa D, Merelli B, Ciano M, Audrito V, Serra S, Buonincontri R, Baroni G, Nassini R, Minocci D, Cattaneo L, Tamborini E, Carobbio A, Rulli E, Deaglio S, Mandalà M.

PD-L1 marks a subset of melanomas with a shorter overall survival and distinct genetic and morphological characteristics.

Ann Oncol. 2014 Dec;25(12):2433-42. doi: 10.1093/annonc/mdu452.

 

Arruga F, Gizdic B, Serra S, Vaisitti T, Ciardullo C, Coscia M, Laurenti L, D'Arena G, Jaksic O, Inghirami G, Rossi D, Gaidano G, Deaglio S.

Functional impact of NOTCH1 mutations in chronic lymphocytic leukemia.

Leukemia. 2014 May;28(5):1060-70. doi: 10.1038/leu.2013.319.

 

Garello F, Stefania R, Aime S, Terreno E, Delli Castelli D.

Successful entrapping of liposomes in glucan particles: an innovative micron-sized carrier to deliver water-soluble molecules.

Mol Pharm. 2014;11(10):3760-5. doi: 10.1021/mp500374f.

For general questions, training and access contact

Dott.ssa Marta Gai

011-6706424