Tracking of Cell Nuclei for Assessment of In Vitro Uptake Kinetics in Ultrasound-Mediated Drug Delivery Using Fibered Confocal Fluorescence Microscopy
- 300 Downloads
Previously, we demonstrated the feasibility to monitor ultrasound-mediated uptake of a cell-impermeable model drug in real time with fibered confocal fluorescence microscopy. Here, we present a complete post-processing methodology, which corrects for cell displacements, to improve the accuracy of pharmacokinetic parameter estimation.
Nucleus detection was performed based on the radial symmetry transform algorithm. Cell tracking used an iterative closest point approach. Pharmacokinetic parameters were calculated by fitting a two-compartment model to the time-intensity curves of individual cells.
Cells were tracked successfully, improving time-intensity curve accuracy and pharmacokinetic parameter estimation. With tracking, 93 % of the 370 nuclei showed a fluorescence signal variation that was well-described by a two-compartment model. In addition, parameter distributions were narrower, thus increasing precision.
Dedicated image analysis was implemented and enabled studying ultrasound-mediated model drug uptake kinetics in hundreds of cells per experiment, using fiber-based confocal fluorescence microscopy.
Key wordsDrug delivery Pharmacokinetic parameters Biological barrier Plasma membrane permeabilization Ultrasound bioeffects Fibered confocal fluorescence microscopy Single-particle tracking
These studies were supported by the EU project SonoDrugs (FP7-NMP4-LA-2008-213706) and ERC project 268906 “Sound Pharma.”
Conflict of Interest
The authors declare that they have no conflict of interest.
Cell-nucleus tracking - monitoring of SYTOX Green cellular uptake. (MP4 4928 kb)
(MP4 2439 kb)
(MP4 4100 kb)
- 10.Tsien RY (2003) Imagining imaging’s future. Nat Cell Biol 5(sup/1):16–21Google Scholar
- 22.Cressie N (1993) Statistics for spatial data. Wiley, Revised EditionGoogle Scholar
- 28.Smal I, Niessen W, Meijering E (2007) Advanced particle filtering for multiple object tracking in dynamic fluorescence microscopy images. Biomedical imaging: From nano to macro. ISBI 2007. 4th IEEE International Symposium on 1048–1051Google Scholar