Application of multipulse excitation to study exocytotic processes in cells stained with acridine orange

Abstract

Mucus secretion is the first-line of defence against the barrage of irritants inhaled into human lungs, but abnormally thick and viscous mucus results in many respiratory diseases. Investigation of processes underlying mucus pathology is hampered, in part, by lack of appropriate experimental tools for labeling and studying mucin granule secretion from live cells with high sensitivity and temporal resolution. Fluorescence spectra and fluorescence lifetime of AO measurements reveal significant changes due to aggregation, and this properties can be useful for determination of mucus expansion. In particular report I introduce an original technique that enchances quality of the exocytotic process study in Calu 3 cells. Multi - pulse excitation significantly increases number of excited aggregates at low concentration of Acridine Orange that reveals possibility to monitor lower concentration than with classic single\npulse excitation. Increase of the signal strength increases sensitivity of measurements that is very important for the detection of small concentrations (up to nanomolars)--Abstract.