Tracking of Cell Nuclei for Assessment of In Vitro Uptake Kinetics in Ultrasound-Mediated Drug Delivery using Fibered Confocal Fluorescence Microscopy

Abstract : Purpose: Previously, we demonstrated the feasibility to monitor in real-time the ultrasound-mediated uptake of a cell-impermeable model drug with fibered confocal fluorescence microscopy. Here we present a complete post-processing methodology in order to improve the accuracy of the measured pharmacokinetic parameters. Procedures: After the detection of the nuclei based on a radial symmetry transform algorithm, a frame-by-frame tracking allowed for the monitoring of each individual uptake. The resulting pharmacokinetic parameters were derived from a two-compartment model. Results: With the tracking, 93% of the 370 nuclei showed a fluorescence signal variation that was well described by a two-compartment model. For each kinetic parameter, the tracking allowed for more homogeneous distributions. Conclusions: This post-processing methodology improved the accuracy of the uptake pharmacokinetic parameters assessed in a cell population. This study extends the proof of concept of designing an in vitro setup for the real-time monitoring of an US-mediated model drug uptake.
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Marc Derieppe, Baudouin Denis de Senneville, Hugo Kuijf, Chrit Moonen, Clemens Bos. Tracking of Cell Nuclei for Assessment of In Vitro Uptake Kinetics in Ultrasound-Mediated Drug Delivery using Fibered Confocal Fluorescence Microscopy. Molecular Imaging and Biology, Springer Verlag, 2014, 16 (5), pp.642-651. ⟨10.1007/s11307-014-0726-3⟩. ⟨hal-00994166v2⟩

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