IMAGING

The facility houses a large variety of systems ranging from basic stereo microscopes, fluorescence widefield and confocal laser scanning microscope to the latest High-Content System in the market by Nikon and structure illumination super-resolution microscopy.

We provide users training and support that range from help with experimental design, image processing and quantitative image analysis, to the establishment, development and coordination of collaboration on specific projects in any field of biological research.

Activity

Our facility is equipped with a tandem scanner filterless confocal Leica SP5, a fully automated time-lapse widefield system (NikonTiE) with CREST spinning disk and super resolution and a variety of light widefield microscopes.

Additionally several image analysis and visualization software packages (Volocity, Metamorph, ImageJ) are available for users to analyze their images.

Our areas of expertise cover:

• Abberior Instruments Stedycon for STED super-resolution microscopy and confocal microscopy.
• Training of basic microscopy techniques including confocal and multicontent.
• Consulting service on immunofluorescence protocols as well on procedures for analyzing and interpreting the results of these methods.
• Multi-color image acquisition and 3D rendering on confocal microscope.
• Super-resolution image acquisition via structure illumination in 3D.
• Live denoise/deconvolution modules in spinning disk confocal acquisition.
• Fast and efficient multi-point and tile-scan image acquisition – from automated image recording routines to complex multi-dimensional high-content screening experiments.
• Time-lapse imaging with reduced photodamage on live cells for several days thanks to LED illumination together with a sCMOS camera for acquisition.
• Developing Macro for automated image analysis.

Team

Equipment

• Confocal Laser Scanning Microscope Leica SP5 with environmental chamber and motorized stage for automated acquisition.
• Fully automated Led-excited multi-porpoise Video-Confocal Nikon TiE microscope with environmental chamber for long time-lapse experiments and multi-channel (up to 24 colors) high-content acquisition in either widefield or spinning disk confocal modality with a specific structure illumination model for super resolution acquisition.
• Inverted widefield Leica microscope with environmental chamber and motorized stage.
• Several widefield/epifluorescence microscopes (inverted and upright) for routine experimentation.
• Dedicated workstations for image analysis equipped with Volocity, NIS-Elements AR, Las-X, ImageJ, Cell Profiler.

Selected publications

• TCTN2: a novel tumor marker with oncogenic properties.
  Cano-Rodriguez D, Campagnoli S, Grandi A, Parri M, Camilli E, Song C, Jin B, Lacombe A, Pierleoni A, Bombaci M, Cordiglieri C, Ruiters MH, Viale G, Terracciano L, Sarmientos P, Abrignani S, Grandi G, Pileri P, Rots MG, Grifantini R.
  Oncotarget. (2017) 24;8 (56):95256-95269
• Platelets from glioblastoma patients promote angiogenesis of tumor endothelial cells and exhibit increased VEGF content and release. 
  Di Vito C, Navone SE, Marfia G, Abdel Hadi L, Mancuso ME, Pecci A, Crisa FM, Berno V, Rampini P, Campanella R, Riboni L.
  Platelets (2017) 28:585-594
  
• Uncontrolled IL-17 Production by Intraepithelial Lymphocytes in a Case of non-IPEX Autoimmune Enteropathy.
  Paroni M, Magarotto A, Tartari S, Nizzoli G, Larghi P, Ercoli G, Gianelli U, Pagani M, Elli L, Abrignani S, Conte D, Geginat J, Caprioli F.
  Clin Transl Gastroenterol (2016) 7:e182
  
• A Myc-driven self-reinforcing regulatory network maintains mouse embryonic stem cell identity.
  Fagnocchi L, Cherubini A, Hatsuda H, Fasciani A, Mazzoleni S, Poli V, Berno V, Rossi RL, Reinbold R, Endele M, Schroeder T, Rocchigiani M, Szkarlat Z, Oliviero S, Dalton S, Zippo A.
  Nat Commun (2016) 7:11903
  
• The Adipose Mesenchymal Stem Cell Secretome Inhibits Inflammatory Responses of Microglia: Evidence for an Involvement of Sphingosine-1-Phosphate Signalling.
  Marfia G, Navone SE, Hadi LA, Paroni M, Berno V, Beretta M, Gualtierotti R, Ingegnoli F, Levi V, Miozzo M, Geginat J, Fassina L, Rampini P, Tremolada C, Riboni L, Campanella R.
  Stem Cells Dev (2016) 25:1095-107
  
• The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma.
  Di Cristofori A, Ferrero S, Bertolini I, Gaudioso G, Russo MV, Berno V, Vanini M, Locatelli M, Zavanone M, Rampini P, Vaccari T, Caroli M, Vaira V.
  Oncotarget (2015) 6:17514-31
  
• NS5A inhibitors impair NS5A-phosphatidylinositol 4-kinase IIIα complex formation and cause a decrease of phosphatidylinositol 4-phosphate and cholesterol levels in hepatitis C virus-associated membranes.
  Reghellin V, Donnici L, Fenu S, Berno V, Calabrese V, Pagani M, Abrignani S, Peri F, De Francesco R, Neddermann P.
  Antimicrob Agents Chemother (2014) 58:7128-40