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Molecular Imaging and Biology

, Volume 16, Issue 5, pp 642–651 | Cite as

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

  • Marc Derieppe
  • Baudouin Denis de Senneville
  • Hugo Kuijf
  • Chrit Moonen
  • Clemens Bos
Research Article

Abstract

Purpose

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.

Procedures

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.

Results

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.

Conclusions

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 words

Drug delivery Pharmacokinetic parameters Biological barrier Plasma membrane permeabilization Ultrasound bioeffects Fibered confocal fluorescence microscopy Single-particle tracking 

Notes

Acknowledgments

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.

Supplementary material

11307_2014_726_MOESM1_ESM.pdf (171 kb)
ESM 1 (PDF 170 kb)
Supplementary data – Videos 1, 2 and 3

Cell-nucleus tracking - monitoring of SYTOX Green cellular uptake. (MP4 4928 kb)

(MP4 2439 kb)

(MP4 4100 kb)

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Copyright information

© World Molecular Imaging Society 2014

Authors and Affiliations

  • Marc Derieppe
    • 1
  • Baudouin Denis de Senneville
    • 1
    • 2
  • Hugo Kuijf
    • 1
  • Chrit Moonen
    • 1
  • Clemens Bos
    • 1
  1. 1.Imaging DivisionUniversity Medical Center UtrechtUtrechtNetherlands
  2. 2.Institut de Mathématiques de BordeauxUMR 5251 CNRS-Université Bordeaux 1-INRIABordeauxFrance

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