Niclas Bengtsson

Gold-Speckled Multimodal Nanoparticles for Noninvasive Bioimaging

In this report the synthesis, characterization, and functional evaluation of a multimodal nanoparticulate contrast agent for noninvasive imaging through both magnetic resonance imaging (MRI) and photoacoustic tomography (PAT) is presented. The nanoparticles described herein enable high resolution and highly sensitive three-dimensional diagnostic imaging through the synergistic coupling of MRI and PAT capabilities. Gadolinium (Gd)-doped gold-speckled silica (GSS) nanoparticles, ranging from 50 to 200 nm, have been prepared in a simple one-pot synthesis using nonionic microemulsions. The photoacoustic signal is generated from a nonuniform, discontinuous gold nanodomains speckled across the silica surface, whereas the MR contrast is provided through Gd incorporated in the silica matrix. The presence of a discontinuous speckled surface, as opposed to a continuous gold shell, allows sufficient bulk water exchange with the Gd ions to generate a strong MR contrast. The dual imaging capabilities of the particles have been demonstrated through in silicio and in vitro methods. The described particles also have the capacity for therapeutic applications including the thermal ablation of tumors through the absorption of irradiated light.

Gadolinium‐Doped Silica Nanoparticles Encapsulating Indocyanine Green for Near Infrared and Magnetic Resonance Imaging

Clinical applications of the indocyanine green (ICG) dye, the only near infrared (NIR) imaging dye approved by the Food and Drug Administration (FDA) in the USA, are limited due to rapid protein binding, fast clearance, and instability in physiologically relevant conditions. Encapsulating ICG in silica particles can enhance its photostability, minimize photobleaching, increase the signal-to-noise (S/N) ratio and enable in vivo studies. Furthermore, a combined magnetic resonance (MR) and NIR imaging particulate can integrate the advantage of high-resolution 3D anatomical imaging with high-sensitivity deep-tissue in-vivo fluorescent imaging. In this report, a novel synthesis technique that can achieve these goals is presented. A reverse-microemulsion-based synthesis protocol is employed to produce 25 nm ICG-doped silica nanoparticles (NPs). The encapsulation of ICG is achieved by manipulating coulombic attractions with bivalent ions and aminated silanes and carrying out silica synthesis in salt-catalyzed, mildly basic pH conditions using dioctyl sulfosuccinate (AOT)/heptane/water microemulsion system. Furthermore, paramagnetic properties are imparted by chelating paramagnetic Gd to the ICG-doped silica NPs. Aqueous ICG-dye-doped silica NPs show increased photostability (over a week) and minimal photobleaching as compared to the dye alone. The MR and optical imaging capabilities of these particles are demonstrated through phantom, in vitro and in vivo experiments. The described particles have the potential to act as theranostic agents by combining photodynamic therapy through the absorption of NIR irradiated light.

lacZ as a genetic reporter for real-time MRI

Molecular imaging based on MRI is currently hampered by the lack of genetic reporters for in vivo imaging. We determined that the commercially available substrate S‐Gal™ can be used to detect genetically engineered β‐galactosidase expressing cells by MRI. The effect and specificity of the reaction between β‐galactosidase and S‐Gal™ on MRI contrast were determined both in vitro and in vivo. β‐galactosidase activity in the presence of S‐Gal™ resulted in enhanced T2 and T*2 MR‐contrast, which was amplified with increasing magnetic field strengths (4.7‐17.6 T) in phantom studies. Using both lacZ+ transgenic animals and lacZ+ tissue transplants, we were able to detect labeled cells in live animals in real time. Similar to phantom studies, detection of the labeled cells/tissues in vivo was enhanced at high magnetic fields. These results demonstrate that the genetic reporter, lacZ, can be used as an in vivo marker gene using high‐field‐strength MRI. Magn Reson Med, 2010.

Ultra-high-field MRI real-time imaging of HSC engraftment of the bone marrow niche

The bone marrow (BM) undergoes extensive remodeling following irradiation damage. A crucial part of restoring homeostasis following irradiation is the ability of hematopoietic stem cells (HSCs) to home to and engraft specialized niches within the BM through a remodeling BM vascular system. Here we show that a combination of ultra-high-field strength magnetic resonance imaging (17.6 T, MRI) coupled with fluorescent microscopy (FLM) serves as a powerful tool for the in vivo imaging of cell homing within the BM. Ultra-high-field MRI can achieve high-resolution three-dimensional (3D) images (28 × 28 × 60 μm3) of the BM in live mice, sufficient to resolve anatomical changes in BM microstructures attributed to radiation damage. Following intra-arterial infusion with dsRed-expressing BM cells, labeled with superparamagnetic iron oxides, both FLM and MRI could be used to follow initial homing and engraftment of donor HSC to a limited number of preferred sites within a few cell diameters of the calcified bone—the endosteal niche. Subsequent histology confirmed the fidelity and accuracy of MRI to create non-invasive, high-resolution 3D images of donor cell engraftment of the BM in living animals at the level of single-cell detection.

Magnetic Resonance Microscopy (MRM) of Single Mammalian Myofibers and Myonuclei

Recently, the first magnetic resonance microscopy (MRM) images at the cellular level in isolated mammalian brain tissues were obtained using microsurface coils. These methods can elucidate the cellular origins of MR signals and describe how these signals change over the course of disease progression and therapy. In this work, we explore the capability of these microimaging techniques to visualize mouse muscle fibers and their nuclei. Isolated myofibers expressing lacZ were imaged with and without a stain for β-galactosidase activity (S-Gal + ferric ammonium citrate) that produces both optical and MR contrast. We found that MRM can be used to image single myofibers with 6-μm resolution. The ability to image single myofibers will serve as a valuable tool to study MR properties attributed to healthy and myopathic cells. The ability to image nuclei tagged with MR/Optical gene markers may also find wide use in cell lineage MRI studies.