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Updated: Apr 18, 2026
A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva
Published on: July 5, 2017
Isolation of microRNA from conjunctival impression cytology
Qistina Pilson1, Caroline A Jefferies2, Joan Ní Gabhann2
1Department of Ophthalmology, Royal College of Surgeons in Ireland, Dublin 2, Ireland; Department of Ophthalmology, Royal Victoria Eye and Ear Hospital, Dublin 2, Ireland.
Abstract:
Impression cytology (IC) is an easy and safe technique that has been used in the past for harvesting epithelial cells from the cornea and conjunctiva for various applications including histology, immunohistology and molecular studies. Previous investigations have shown the usage of different types of membranes for the purpose of investigating pathophysiology and staging of diseases. This contributes to a better understanding of ocular surface conditions and helps to provide information for diagnosis, therapeutic options and prognosis. Recently, there has been a shift of focus in research towards understanding the contribution of microRNAs (miRs) to ocular disease. Thus far, impression cytology has been explored for measuring gene expression but not for quantifying miR expression. This study describes how miRs and mRNA can be isolated from conjunctival epithelial cells obtained by impression cytology and determines the optimum membrane and technique for this purpose. The IC technique was optimized using Biopore, Immobilon-P(SQ) and Millicell Hanging Cell Culture Insert membranes on healthy controls. miRs and mRNAs were isolated from the conjunctival epithelial cells (CEC) obtained and measured. Biopore membrane provided the optimum yield of miRs (38.8 ng/μL ± 10.8) and mRNA (155.3 ng/μL ± 20.1) as well as subjectively found to be best tolerated with minimum discomfort. Appreciable levels of miRs and mRNAs were detected from the CEC from healthy controls, confirming that it is possible to isolate miR and mRNA from CEC. Here, we give a detailed description of the application of conjunctival impression cytology to isolate miRs and the convenience of the technique by using the best membrane available. This method can be readily adopted in both clinical and laboratory settings. This technique will facilitate the measurement of miRs to improve our understanding of the pathogenesis of ocular surface conditions as well as potentially identifying novel therapeutic targets.
Insights
Impression cytology effectively isolates microRNAs (miRs) and mRNA from conjunctival cells. The Biopore membrane offers optimal yield and patient comfort for ocular surface disease research.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Impression cytology (IC) is a safe technique for collecting ocular surface cells for analysis.
- Previous research utilized IC for histology and gene expression studies.
- Recent focus is on microRNA (miR) contributions to ocular diseases, but IC's utility for miR quantification is unexplored.
Purpose of the Study:
- To describe a method for isolating miRs and mRNA from conjunctival epithelial cells (CECs) using IC.
- To determine the optimal membrane and technique for miR and mRNA isolation via IC.
- To validate the feasibility of quantifying miRs and mRNA from CECs obtained through IC.
Main Methods:
- Optimized impression cytology technique using Biopore, Immobilon-P(SQ), and Millicell membranes on healthy controls.
- Isolated and quantified miRs and mRNAs from conjunctival epithelial cells (CECs) collected via IC.
- Evaluated membrane performance based on yield and subjective patient comfort.
Main Results:
- Biopore membrane yielded the highest amounts of miRs (38.8 ng/μL ± 10.8) and mRNA (155.3 ng/μL ± 20.1).
- Biopore membrane was subjectively found to be best tolerated with minimal patient discomfort.
- Feasible isolation and detection of miRs and mRNAs from CECs were confirmed.
Conclusions:
- Conjunctival impression cytology with the Biopore membrane is an effective method for isolating miRs and mRNA.
- This optimized IC technique can be readily adopted in clinical and laboratory settings for ocular surface research.
- This approach facilitates miR measurement, enhancing understanding of ocular surface disease pathogenesis and potentially identifying therapeutic targets.
