Optimising the method for isolating ocular surface microRNA using impression cytology

Purpose Conjunctival impression cytology is a reliable method to obtain epithelial cells from the ocular surface. Its application has broadened our knowledge of surface disease, paving way for greater understanding pathogenesis new treatment options and personalised medicine. Herein we compare most commonly used materials optimise isolating microRNA (miR) conjunctiva. Methods MiR yield 12 mm PTFE 13 PES in free membrane semicircle form were compared (n = 3). Next single Eyeprim applicator 8). MiRs isolated using miRNeasy kit (QIAGEN GmbH Hilden Germany) concentration was determined on Nanodrop ND-1000 Spectrophotometer (Thermo Fisher Scientific Wilmington DE). In second experiment, subjective discomfort provoked by each technique also SANDE immediately 2 min after sampling. Results had significantly higher miR vs PTFE; 131.3 ng/µl±8.1 32 ng/µl±9.9 ng/µl p 0.0002. experiment comparable Eyeprim; 48.7 ng/µl±24.4 46.9 ng/µl±19.2, 0.88. measured participant this experiment. Regarding intensity tolerable at point sampling; 0.1± 0.3 4.5±2.8 Eyeprim, 0.0016. At min, 0.1 ± 3.3 3.22 0.03. Conclusions Our results confirm that greatly superior its harvesting ability which previously optimised studies Millicell Biopore. Furthermore, commercially designed while being better tolerated. We recommend use 0.2 µm pore design harvest characterise expression conjunctival cells. This simple, inexpensive can easily be implemented clinical research setting investigate disease.