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Updated: Mar 15, 2026
Impression Cytology of the Lid Wiper Area
Published on: August 9, 2016
Impression Cytology of the Lid Wiper Area
Alex Muntz1, Kevin van Doorn2, Lakshman N Subbaraman2
1Centre for Contact Lens Research, University of Waterloo; amuntz@uwaterloo.ca.
Insights
This study optimizes impression cytology (IC) for analyzing the lid wiper (LW) area, revealing cellular details and metabolic activity. The enhanced method offers higher sensitivity for studying conditions like dry eye and contact lens wear.
Area of Science:
- Ophthalmology
- Cell Biology
- Cytology
Background:
- Limited understanding of the lid wiper (LW) area's cellular anatomy.
- Previous cytological studies of the LW area are scarce.
Purpose of the Study:
- To optimize a method for collecting, staining, and imaging cells from the human inner eyelid's LW region using impression cytology (IC).
- To establish a sensitive technique for assessing cellular morphology and metabolic activity in the LW area.
Main Methods:
- Cells collected from the inner eyelid using hydrophilic polytetrafluoroethylene (PTFE) membranes.
- Staining with cytological and immunocytochemical dyes to identify cell types, mucins, nuclei, keratinization, esterase activity, and membrane integrity.
- Confocal scanning laser microscopy and image stitching to create high-resolution images of the IC span.
Main Results:
- Demonstrated higher sensitivity of IC for LW area analysis compared to previous reports.
- Successfully visualized goblet cells, mucins, cell nuclei, keratinization, esterase activity, and compromised cell membranes.
- Developed a protocol for high-resolution imaging of the entire IC span.
Conclusions:
- The optimized IC protocol provides a sensitive tool for examining the lid wiper area's cellular details and metabolic state.
- This method is suitable for future research on contact lens wearers, dry eye patients, and lid wiper epitheliopathy.
- This study introduces a novel application of fluorescent dyes for imaging LW IC membranes.
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.
