Biology subjects

Bismuth, M.

Publications and source records attributed to Bismuth, M..

5 recordsLinked to original sources

bioRxiv2026

Nanobubble-Based Ultrasound Localization Microscopy through Interactive Adaptive Processing

This study presents the use of sub-micron nanobubbles (NBs) as contrast agents for ultrasound localization microscopy (ULM), a super-resolution imaging technique that visualizes microvascular structure and flow beyond the acoustic diffraction limit. While ULM has traditionally relied on micron-sized microbubbles (MBs), the reduced dimensions and prolonged circulation times of NBs make them attractive candidates for localization-based imaging. However, their weaker acoustic responses present significant challenges for reliable detection and tracking. To address this challenge, we developed the ULM Master GUI, an interactive framework for optimization of the complete ULM processing pipeline. Using custom ultrasound-compatible wall-less gelatin flow phantoms containing vessel-mimicking channels and bifurcations ranging from 100 to 500 m, we demonstrate that NB-based ULM achieves velocity reconstruction and flow partitioning measurements comparable to conventional MB-based ULM. Across all investigated geometries, NBs faithfully reproduced the underlying flow patterns and hemodynamic behavior despite their substantially reduced acoustic scattering. These findings establish the feasibility of NB-based ULM, expand the range of contrast agents available for localization microscopy, and provide a foundation for future super-resolution ultrasound imaging using nanoscale acoustic contrast agents. The ULM processing GUI is publicly available at https://github.com/grisha1998/ulm-super-resolution-toolbox.

bioengineering↗

bioRxiv2026

Super-Resolution Macrophage Imaging via Ultrasound Localization Microscopy and Blinking Nanodroplets

Tracking immune cells deep within living tissue remains a fundamental challenge due to the diffraction-limited resolution of ultrasound imaging and the inability to resolve dense cellular populations. Here, we introduce an intracellular super-resolution ultrasound imaging framework based on stochastic phase-changing nanodroplets (NDs) and ultrasound localization microscopy (ULM). We engineer [~]170 nm perfluorocarbon NDs that undergo reversible, stochastic liquid-gas transitions under acoustic excitation, generating temporally sparse "blinking" signals. Leveraging the intrinsic endocytic activity of macrophages, these NDs are internalized, enabling intracellular contrast generation independent of vascular flow. We validate this approach across imaging scales, from controlled phantoms and in vitro systems to in vivo tumor models, demonstrating robust intracellular blinking, high cell viability, and consistent super-resolution reconstruction in dense cellular environments. The stochastic blinking of internalized NDs provides the temporal separation required to localize individual sources, overcoming a central limitation of conventional ULM. Following systemic administration, ND-labeled macrophages are tracked in vivo after homing to the liver, where super-resolution ULM resolves cellular distributions with a spatial resolution of 26.3 {+/-} 3.2 {micro}m, corresponding to a 6.1-fold improvement over diffraction-limited imaging. This work establishes a previously unexplored paradigm for ultrasound-based intracellular super-resolution imaging, enabling non-invasive visualization of immune cell organization in deep tissue. By introducing spatiotemporally programmable intracellular contrast, this approach expands ultrasound beyond vascular imaging toward functional cellular imaging, with broad implications for immunology, diagnostics, and cell-based therapies.

bioengineering↗

bioRxiv2026

Acoustically activatable drug-loaded nanodroplets for mechanochemical therapy in solid tumors

Stimulus-responsive nanomedicines promise spatiotemporally controlled therapy, yet most systems rely on passive delivery and lack precise, externally programmable activation while maintaining clinical compatibility. Here we engineer sub-200 nm, perfluorocarbon (PFC)-core nanodroplets (NDs) that integrate efficient core drug loading, physiological stability, and acoustically programmable activation within a single nanoscale agent. These NDs are fabricated using microfluidic nanoassembly to achieve controlled size and composition, and are designed to encapsulate fluorinated payloads directly within the liquid core. Upon exposure to a sequential dual-frequency ultrasound (US) paradigm, the NDs undergo acoustic droplet vaporization followed by low-frequency cavitation, enabling spatially confined mechanical disruption and on-demand payload release within clinically relevant acoustic limits. These properties are engineered to overcome physicochemical barriers in solid tumors, including dense extracellular matrix and restricted drug penetration. This approach achieves enhanced payload release and induces potent mechanochemical cytotoxicity in vitro. In vivo, NDs exhibit prolonged circulation and tumor accumulation, while US activation drives substantial tissue fractionation, control drug release, and increases subsequent nanoparticle uptake. When applied to a solid tumor model, this combined mechanochemical strategy improves tumor control and significantly extends survival compared to either modality alone. These acoustically activatable NDs provide a versatile system for stimulus-responsive, site-targeted drug delivery and mechanical tumor disruption, with strong potential for clinical translation. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=99 SRC="FIGDIR/small/719550v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@9c07f8org.highwire.dtl.DTLVardef@1cf355aorg.highwire.dtl.DTLVardef@b7afd1org.highwire.dtl.DTLVardef@177fa1a_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2026

Cryoablation temperature monitoring with dense ultrasonic speed-of-sound shift imaging

Accurate temperature monitoring during cryoablation, a minimally invasive technique that destroys tissue locally by forming an ice ball around an inserted cryoprobe, is vital for achieving complete ablation while protecting surrounding tissue. We present a dense slowness-shift imaging method that estimates local speed-of-sound changes from ultrasound B-mode images using optical flow. This single-transducer, image-based approach enables mapping of spatial temperature change without requiring additional hardware. Cryoablation experiments in a tissue-mimicking phantom and ex vivo turkey breast demonstrated that slowness deviation increases with decreasing temperature. In the phantom, the dependence was linear (a = -20.70 s{middle dot}m-1C{degrees}-1), while in turkey breast it presented an exponential relationship (t = 34.04xexp(0.075(-{Delta}T)) s{middle dot}m-1C{degrees}-1). The algorithm detected sub-degree temperature variations and accurately tracked cooling down to -39.4 {+/-} 5.6 {degrees}C. This work demonstrates the feasibility of ultrasound-based, noninvasive temperature monitoring during cryoablation, providing a scalable, real-time alternative to existing invasive or high-cost thermal assessment techniques.

biophysics↗

bioRxiv2025

Low-frequency ultrasound-mediated blood-brain barrier opening enables non-invasive lipid nanoparticle RNA delivery to glioblastoma

Ionizable Lipid Nanoparticles (LNPs) are an FDA-approved non-viral RNA delivery system, though their use for brain therapy is restricted by the blood-brain barrier (BBB). Focused ultrasound combined with microbubbles can disrupt the BBB, but delivering large particles requires balancing increased peak negative pressures while maintaining microvascular integrity. Herein, we optimized low-frequency ultrasound parameters to induce high-amplitude microbubble oscillations, enabling the safe delivery of LNPs across the BBB. First, BBB opening was assessed at different frequencies (850, 250, and 80 kHz) and pressures by monitoring the extravasation of Evans blue (~1 kDa). Next, the delivery of 4, 70, and 150 kDa Dextrans, LNPs entrapping Cy5-siRNAs (~70 nm in diameter), and LNPs entrapping mRNA (~100 nm in diameter) was evaluated via microscopy and bioluminescence. In a glioblastoma syngeneic mouse model, siRNA-Cy5-LNP was successfully delivered. A frequency of 850 kHz and 125 kPa pressure induced safe BBB opening, enabling delivery of both small molecules and LNPs. In healthy brains, LNP entrapping siRNAs delivery increased 10-fold compared to controls, and LNPs with mRNAs showed a 12-fold increase in bioluminescence after 24 hours. In glioblastoma tumors, LNPs with siRNAs delivery resulted in a 6.7-fold increase in fluorescence. This study paves the way for non-invasive LNP delivery to the brain, offering a versatile platform for brain therapies. TOC graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=76 SRC="FIGDIR/small/634427v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@cad2bdorg.highwire.dtl.DTLVardef@1d1936corg.highwire.dtl.DTLVardef@259d83org.highwire.dtl.DTLVardef@1883aed_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗