1 Department of Physics, Concordia University, Montreal, Quebec H4B 1R6, Canada.
2 Department of Physics, Concordia University, Montreal, Quebec H4B 1R6, Canada; Department of Biology, Concordia University, Montreal, Quebec H4B 1R6, Canada. Electronic address: brandon.helfield@concordia.ca.

Phospholipid encapsulated ultrasound contrast agents have proven to be a powerful addition in diagnostic imaging and show emerging applications in targeted therapy due to their resonant and nonlinear scattering. Microbubble response is affected by their intrinsic (e.g. bubble size, encapsulation physics) and extrinsic (e.g. boundaries) factors. One of the major intrinsic factors at play affecting microbubble vibration dynamics is the initial phospholipid packing of the lipid encapsulation. Here, we examine how the initial phospholipid packing affects the subharmonic response of either individual or a system of two closely-placed microbubbles. We employ a finite element model to investigate the change in subharmonic resonance under 'small' and 'large' radial excursions. For microbubbles ranging between 1.5 and 2.5 µm in diameter and in its elastic state (s₀ = 0.01 N/m), we demonstrate up to a 10 % shift towards lower frequencies in the peak subharmonic response as the radial excursion increases. However, for a bubble initially in its buckled state (s₀ = 0 N/m), we observe a maximum shift of 8 % towards higher frequencies as the radial excursion increases over the same range of bubble sizes - the opposite trend. We studied the same scenario for a system of two individual microbubbles for which we saw similar results. For microbubbles that are initially in their elastic state, in both cases of a) two identically sized bubbles and b) a bubble in proximity to a smaller bubble, we observed a 6 % and 9 % shift towards lower frequencies respectively; while in the case of a neighboring larger bubble no change in subharmonic resonance frequency was observed. Microbubbles that are initially in a buckled state exert no change, 5 % and 19 % shift towards higher frequencies, in two-bubble systems consisting of a) same-size, b) smaller, and c) larger neighboring bubble respectively. Furthermore, we examined the effect of two adjacent bubbles with non-equal initial phospholipid states. The results presented here have important implications in ultrasound contrast agent applications.