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Quantitative evaluation of ultrasound-mediated cellular uptake of a fluorescent model drug
Matthieu Lepetit-Coiffé1, Anna Yudina, Christel Poujol
1Laboratoire IMF UMR 5231 CNRS, Université Bordeaux 2, Bordeaux, France, matlepco@hotmail.com.
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.
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