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Evaluation Of Ocular Surface Disorders: A New Diagnostic Tool Based On Impression Cytology And Confocal Laser Scanning Microscopy
  1. Vanessa Barbaro1,*,
  2. Stefano Ferrari1,
  3. Adriano Fasolo1,
  4. Emilio Pedrotti2,
  5. Giorgio Marchini2,
  6. Arianna Sbabo2,
  7. Nicola Nettis1,
  8. Diego Ponzin1,
  9. Enzo Di Iorio1
  1. 1 The Veneto Eye Bank Foundation, Italy;
  2. 2 Eye Clinic, Department of Neurological and Visual Sciences, University of Verona, Italy
  1. To whom correspondence should be addressed. E-mail: vanessa.barbaro{at}


Aim: To provide a new tool for the evaluation of altered ocular surfaces by using a combination of impression cytology, laser scanning confocal microscopy and advanced image analysis.

Methods: The expression of keratin 3 (K3), keratin 12 (K12), keratin 19 (K19) and mucin 1 (MUC1) was analysed through immunofluorescence on both histological sections of conjunctiva, limbus and cornea and impression cytology specimens from six healthy normal subjects (12 eyes) and twelve patients diagnosed with chronic ocular surface disorders. Levels of fluorescence expression of the different markers were quantified through Quantitative Fluorescence ImmunoHistoChemistry (Q-FIHC).

Results: Impression cytology specimens from normal and diseased ocular surfaces showed distinct expression patterns for K12 and MUC1. Healthy corneas expressed only K12 (but not MUC1) while conjunctivalized corneas from patients with LSC were characterized by the presence of MUC1 and the disappearance of K12. Similar clear-cut results were not seen with the traditional pair of markers K3/K19, which showed lack of specificity and overlapping signals in cornea and conjunctiva impression cytology specimens.

Conclusions: The ability of K12 and of the antibody used to detect MUC1 to discriminate clearly between limbus/cornea and conjunctiva in impression cytology specimens could become a valuable diagnostic and clinical monitoring tool for ophthalmologists in order to evaluate alterations of the ocular surface and the grading of LSC deficiency.

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      Vanessa Barbaro Stefano Ferrari Adriano Fasolo Emilio Pedrotti Giorgio Marchini Arianna Sbabo Nicola Nettis Diego Ponzin Enzo Di Iorio