Quantitative evaluation of ultrasound-mediated cellular uptake of a fluorescent model drug

Mol Imaging Biol. 2013 Oct;15(5):523-33. doi: 10.1007/s11307-013-0615-1.

Abstract

Purpose: This study aims to quantitatively analyze cellular uptake following local ultrasound (US)-mediated cell permeabilization.

Procedures: A 2 μM cell-impermeable dye Sytox Green was co-injected with 3 × 10(7) microbubbles in the presence of C6 rat glioblastoma cell monolayer in total volume of 10 ml. A 5.8-mm diameter mono-element US transducer was positioned at a distance of 8 mm to the Opticell® membrane. Acoustical pressure of pulsed US was varied from 0.62 MPa peak-to-peak (p-p) to 1.25 MPa p-p. Large field of view (FOV = 15 × 15 mm) 22 × 22 mosaic acquisitions were done under epifluorescence Leica DMR microscope and analyzed in Metamorph software to evaluate cell density as well as model drug uptake percentage.

Results: The size of acoustical field of the transducer closely matches the spatial pattern of the model drug internalized into the cells by US. Maximum of uptake percentage (42 ± 15 %) was found at 0.88 MPa p-p.

Conclusions: Spatial aspect of US-mediated model drug uptake has been quantitatively evaluated on adherent cells using robust 2D-mapping approach.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Endocytosis*
  • Glioblastoma / metabolism*
  • Image Processing, Computer-Assisted
  • Microscopy, Fluorescence
  • Molecular Imaging
  • Organic Chemicals / metabolism
  • Pharmaceutical Preparations / metabolism*
  • Rats
  • Ultrasonics

Substances

  • Organic Chemicals
  • Pharmaceutical Preparations
  • SYTOX Green