Analysis of fluorescent images of respiratory epithelial cells from healthy donors

We applied automated image analysis to systematically examine the influence of samples’ storage conditions on the specificity of IF staining with eight polyclonal antibodies routinely used for diagnostics of primary ciliary dyskinesia, PCD. We used antibodies targeting epitopes of proteins representing various axoneme structural complexes. Seven various combinations of temperatures and storage times were tested to mimic the procedures of handling epithelial brushing on glass slides: at the clinic, during transport, or after reception at the diagnostic laboratory. Our study revealed that proper slide storage conditions are essential for the reliable PCD diagnosis via IF staining. Storage temperatures tested and their symbols used in the data files: Condition symbol Group Conditions (temperature and duration time) 1 -80_28d (best conditions) -80°C 28 days 3 -20_28d -20°C28 days 4 -20_8w -20°C 8 weeks 5 -20/RT/-80 -20°C 11 days RT 3 days -80°C 14 days 6 4/RT/-80 4 °C 11 days RT 3 days -80°C 14 days 7 RT/RT/-80 RT 11 days RT 3 days -80°C 14 days 2 RT_28d (worst conditions) RT 28 days