title: Impression Cytology of the Lid Wiper Area.
abstract: Few reports on the cellular anatomy of the lid wiper (LW) area of the inner eyelid exist and only one report makes use of cytological methods. The optimization of a method of collecting, staining and imaging cells from the LW region using impression cytology (IC) is described in this study. Cells are collected from the inner surface of the upper eyelid of human subjects using hydrophilic polytetrafluoroethylene (PTFE) membranes, and stained with cytological dyes to reveal the presence of goblet cells, mucins, cell nuclei and various degrees of pre- and para-keratinization. Immunocytochemical dyes show cell esterase activity and compromised cell membranes by the use of a confocal scanning laser microscope. Up to 100 microscopic digital images are captured for each sample and stitched into a high-resolution, large scale image of the entire IC span. We demonstrate a higher sensitivity of IC than reported before, appropriate for identifying cellular morphologies and metabolic activity in the LW area. To our knowledge, this is the first time this selection of fluorescent dyes was used to image LW IC membranes. This protocol will be effective in future studies to reveal undocumented details of the LW area, such as assessing cellular particularities of contact lens wearers or patients with dry eye or lid wiper epitheliopathy.
authors: Centre for Contact Lens Research, University of Waterloo; amuntz@uwaterloo.ca.
authors: Alex
authors: A
authors: Muntz
authors: Centre for Contact Lens Research, University of Waterloo.
authors: Kevin
authors: K
authors: van Doorn
authors: Centre for Contact Lens Research, University of Waterloo.
authors: Lakshman N
authors: LN
authors: Subbaraman
authors: Centre for Contact Lens Research, University of Waterloo.
authors: Lyndon W
authors: LW
authors: Jones
mesh_terms: D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry; D003228:Conjunctiva / Q000166:cytology*; D015352:Dry Eye Syndromes / Q000000981:diagnostic imaging / Q000473:pathology; D004950:Esterases / Q000032:analysis; D005143:Eyelids / Q000166:cytology* / Q000000981:diagnostic imaging; D005456:Fluorescent Dyes; D020397:Goblet Cells / Q000166:cytology; D006801:Humans; D008567:Membranes, Artificial; D018613:Microscopy, Confocal; D009077:Mucins / Q000032:analysis; D011138:Polytetrafluoroethylene; D012919:Social Behavior; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D009077:Mucins; D011138:Polytetrafluoroethylene; D004950:Esterases