IMAGING

The facility houses a variety of fluorescence widefield and confocla systems and confocal laser scanning microscopes, plus a super-resolution 3-channel STED instrument and a versitile customized high-content system with spinning disk confocal and structure illumination super-resolution microscopy coupled with 16-channels widefield detection.

Activity

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.

Our areas of expertise cover:

• Training of basic microscopy techniques including super-resolution, 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.
• Super-resolution image acquisition via structure illumination and STED.
• Application of deconvolution modules and ad-hoc implemented processing techniques.
• Fast and efficient multi-point and tile-scan image acquisition – from automated image recording routines to complex multi-dimensional high-content screening experiments.
• Long-duration stable time-lapse imaging for 2D and 3D cellular models
• Highly dynamic, high resolution fast time-laspse imaging on 2D cellular models.
• Development of complex automated image analysis with segmentation pipelines ad-hoc implemented upon user-need.

Team

Equipment

• Abberior Stedycon for 4-channel confocal microscopy and STED 3 channel super-resolution
• Confocal Laser Scanning Microscope Leica SP5 with environmental chamber and motorized stage for automated acquisition.
• Fully automated Lase-excited and Led-excited multi-porpoise
• Video-Confocal Nikon TiE microscope with environmental chamber for long time-lapse experiments and multi-channel high-content acquisition in either widefield or spinning disk confocal modality, with 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 NIS-Elements AR, Las-X, ImageJ, Cell Profiler, iLastik, qPath, Volocity.

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