Understanding
the pressure dependence of the nonlinear behavior
of ultrasonically excited phospholipid-stabilized nanobubbles (NBs)
is important for optimizing ultrasound exposure parameters for implementations
of contrast enhanced ultrasound, critical to molecular imaging. The
viscoelastic properties of the shell can be controlled by the introduction
of membrane additives, such as propylene glycol as a membrane softener
or glycerol as a membrane stiffener. We report on the production of
high-yield NBs with narrow dispersity and different shell properties.
Through precise control over size and shell structure, we show how
these shell components interact with the phospholipid membrane, change
their structure, affect their viscoelastic properties, and consequently
change their acoustic response. A two-photon microscopy technique
through a polarity-sensitive fluorescent dye, C-laurdan, was utilized
to gain insights on the effect of membrane additives to the membrane
structure. We report how the shell stiffness of NBs affects the pressure
threshold (
P
t
) for the sudden amplification
in the scattered acoustic signal from NBs. For narrow size NBs with
200 nm mean size, we find
P
t
to be between
123 and 245 kPa for the NBs with the most flexible membrane as assessed
using C-Laurdan, 465–588 kPa for the NBs with intermediate
stiffness, and 588–710 kPa for the NBs with stiff membranes.
Numerical simulations of the NB dynamics are in good agreement with
the experimental observations, confirming the dependence of acoustic
response to shell properties, thereby substantiating further the development
in engineering the shell of ultrasound contrast agents. The viscoelastic-dependent
threshold behavior can be utilized for significantly and selectively
enhancing the diagnostic and therapeutic ultrasound applications of
potent narrow size NBs.