title: An open source based high content screening method for cell biology laboratories investigating cell spreading and adhesion.
abstract: BACKGROUND
  Adhesion dependent mechanisms are increasingly recognized to be important for a wide range of biological processes, diseases and therapeutics. This has led to a rising demand of pharmaceutical modulators. However, most currently available adhesion assays are time consuming and/or lack sensitivity and reproducibility or depend on specialized and expensive equipment often only available at screening facilities. Thus, rapid and economical high-content screening approaches are urgently needed.
  
  
  RESULTS
  We established a fully open source high-content screening method for identifying modulators of adhesion. We successfully used this method to detect small molecules that are able to influence cell adhesion and cell spreading of Swiss-3T3 fibroblasts in general and/or specifically counteract Nogo-A-Δ20-induced inhibition of adhesion and cell spreading. The tricyclic anti-depressant clomipramine hydrochloride was shown to not only inhibit Nogo-A-Δ20-induced cell spreading inhibition in 3T3 fibroblasts but also to promote growth and counteract neurite outgrowth inhibition in highly purified primary neurons isolated from rat cerebellum.
  
  
  CONCLUSIONS
  We have developed and validated a high content screening approach that can be used in any ordinarily equipped cell biology laboratory employing exclusively freely available open-source software in order to find novel modulators of adhesion and cell spreading. The versatility and adjustability of the whole screening method will enable not only centers specialized in high-throughput screens but most importantly also labs not routinely employing screens in their daily work routine to investigate the effects of a wide range of different compounds or siRNAs on adhesion and adhesion-modulating molecules.
authors.affiliation: Brain Research Institute, University of Zurich and Department of Health Sciences and Technology, ETH Zurich, Zurich, Switzerland.
authors.forename: Andre
authors.initials: A
authors.lastname: Schmandke
authors.forename: Antonio
authors.initials: A
authors.lastname: Schmandke
authors.forename: Maurianne A
authors.initials: MA
authors.lastname: Pietro
authors.forename: Martin E
authors.initials: ME
authors.lastname: Schwab
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002478:Cells, Cultured; D022082:Cellular Structures / Q000378:metabolism / Q000502:physiology; D002531:Cerebellum / Q000378:metabolism / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology; D051379:Mice; D009185:Myelin Proteins / Q000378:metabolism*; D016501:Neurites / Q000378:metabolism / Q000502:physiology; D000070798:Nogo Proteins; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015815:Cell Adhesion Molecules; D009185:Myelin Proteins; D000070798:Nogo Proteins; C000603201:Rtn4 protein, mouse; C000603197:Rtn4 protein, rat