Vesselj: A new tool for semiautomatic measurement of corneal neovascularization

Alessandro Rabiolo, Fabio Bignami, Paolo Rama, Giulio Ferrari

Research output: Contribution to journalArticlepeer-review


PURPOSE. To quantify blood and lymph angiogenesis in mouse corneal flat mounts by means of a novel plug-in for ImageJ, called VesselJ, based on a dynamic threshold algorithm. METHODS. Corneal neovascularization (CNV) was induced in the right corneas of 20 C57BL6/N mice by means of alkali burn (n = 10) or intrastromal sutures (n = 10). All corneal flat mounts were stained for blood vessels with CD31 and for lymphatics with LYVE1. Three independent operators measured blood and lymphatic CNV with both a published manual method (mCNV) and VesselJ (automatic method; aCNV). RESULTS. Both methods showed a strong reliability, defined as intraclass correlation coefficient (ICC) > 0.90, in quantifying hemangiogenesis for sutures and alkali burn. However, reliability of lymphatic mCNV varied from moderate in alkali burn (ICC: 0.700) to poor in sutures (ICC: 0.415), whereas it remained high in aCNV (alkali ICC: 0.996; sutures ICC: 0.959). Among sutures, a significant correlation between mCNV and aCNV was found among all the three operators for blood vessels and just for one operator for lymphatic vessels (P <0.001). In the alkali burn model, correlation between blood mCNV and aCNV was significant for all operators after excluding three noisy flat mounts (P <0.001), whereas no significant correlation was seen for lymphatic vessels. CONCLUSIONS. VesselJ is a semiautomatic, reliable, and fast method to quantify corneal hem-and lymphangiogenesis in corneal flat mounts. VesselJ can be easily used in the sutures model; it should be applied to other models (e.g., alkali burn) only after checking for background hyperfluorescence.

Original languageEnglish
Pages (from-to)8199-8206
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Issue number13
Publication statusPublished - Dec 1 2015


  • Blood vessels
  • Corneal neovascularization
  • ImageJ
  • Lymphatics
  • Plug-in

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience


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