Characterizing ultrasound contrast agents (UCAs) involve measuring the size and population distribution. However, these instruments do not allow for characterization of shell properties, which are important for (1) stability to administration and circulation throughout the vasculature; (2) UCA response to ultrasound; and (3) conjugating ligands for molecular imaging. Thus it is critical to understand the physical and rheological properties of shells. We previously developed a light scattering technique to characterize the shell properties of UCAs [Guan and Matula, JASA, vol. 116(5), 2004; Tu, et al., IEEE Trans. Ultrason., Ferroelec., and Freq. Control, vol. 58(5), 2011]. The most recent manifestation involves a flow cytometer modified with a custom square quartz flow cell in place of the standard nozzle and fluid jet. Acoustic coupling to the carrier sheath fluid and UCA samples occurred through a PZT bonded to one side of the flow cell. The PZT-driven UCA oscillations were processed and fitted to the Marmottant UCA model. Shell properties for UCAs (including Definity, Optison, SonoVue, and even homemade bubbles) were determined. The focus of this talk will be on pressure calibration and additional measurements of unpublished data from Optison and homemade bubbles.

Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled using Mie theory, and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures (from 95 to 333 kPa) by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters.

Flow cytometry concepts are modified to enable dynamic characterizations of particles to be obtained using optical scattering data. Particles in flow will be introduced into a sample volume. Light scattered by a particle in the sample volume is collected and analyzed. What differentiates the concepts disclosed herein from conventional flow cytometry is the use of an acoustic source that is disposed to direct acoustic energy into the sample volume. As the particle passes through the sample volume, it responds to the acoustic energy, causing changes in the light scattered by the particle. Those changes, which are not measured during conventional flow cytometry, can be analyzed to determine additional physical properties of the particle.