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

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.

Rapid motion correction in MR-guided high-intensity focused ultrasound heating using real-time ultrasound echo information.

The objective of this study was to evaluate the feasibility of integrating real-time ultrasound echo guidance in MR-guided high-intensity focused ultrasound (HIFU) heating of mobile targets in order to reduce latency between displacement analysis and HIFU treatment. Experiments on a moving phantom were carried out with MRI-guided HIFU during continuous one-dimensional ultrasound echo detection using separate HIFU and ultrasound imaging transducers. Excellent correspondence was found between MR- and ultrasound-detected displacements. Real-time ultrasound echo-based target tracking during MR-guided HIFU heating is shown with the dimensions of the heated area similar to those obtained for a static target. This work demonstrates that the combination of the two modalities opens up perspectives for motion correction in MRI-guided HIFU with negligible latency.

Non-invasive determination of tissue thermal parameters from high intensity focused ultrasound treatment monitored by volumetric MRI thermometry.

A method is proposed for estimating the perfusion rate, thermal diffusivity, and the absorption coefficient that influence the local temperature during high intensity focused ultrasound (HIFU) thermotherapy procedures. For this purpose, HIFU heating experiments (N = 100) were performed ex vivo on perfused porcine kidney (N = 5) under different flow conditions. The resulting spatio-temporal temperature variations were measured non-invasively by rapid volumetric MR-temperature imaging. The bio-heat transfer (BHT) model was adapted to describe the spatio-temporal evolution of tissue temperature in the cortex. Absorption and perfusion coefficients were determined by fitting the integrated thermal load (spatial integration of the thermal maps) curves in time with an analytical solution of the BHT equation proposed for single point HIFU heating. Thermal diffusivity was determined independently by analyzing the spatial spread of the temperature in time during the cooling period. Absorption coefficient and thermal diffusivity were found to be independent of flow, with mean and average values of 11.0 +/- 1.85 mm(3) x K x J(-1) and 0.172 +/- 0.003 mm(2) x s(-1), respectively. A linear dependence of the calculated perfusion rate with flow was observed with a slope of 9.20 +/- 0.75 mm(-3). The perfusion was found to act as a scaling term with respect to temperature but with no effect on the spatial spread of temperature which only depends on the thermal diffusivity. All results were in excellent agreement with the BHT model, indicating that this model is suitable to predict the evolution of temperature in perfused organs. This quantitative approach allows for determination of tissue thermal parameters with excellent precision (within 10%) and may thus help in quantifying the influence of perfusion during MR guided high intensity focused ultrasound (MRgHIFU).

Three-dimensional spatial and temporal temperature control with MR thermometry-guided focused ultrasound (MRgHIFU).

High-intensity focused ultrasound (HIFU) is an efficient noninvasive technique for local heating. Using MRI thermal maps, a proportional, integral, and derivative (PID) automatic temperature control was previously applied at the focal point, or at several points within a plane perpendicular to the beam axis using a multispiral focal point trajectory. This study presents a flexible and rapid method to extend the spatial PID temperature control to three dimensions during each MR dynamic. The temperature in the complete volume is regulated by taking into account the overlap effect of nearby sonication points, which tends to enlarge the heated area along the beam axis. Volumetric temperature control in vitro in gel and in vivo in rabbit leg muscle was shown to provide temperature control with a precision close to that of the temperature MRI measurements. The proposed temperature control ensures heating throughout the volume of interest of up to 1 ml composed of 287 voxels with 95% of the energy deposited within its boundaries and reducing the typical average temperature overshoot to 1 degrees C.