Plasmonic gold nanoshell (GNS) probe penetrates into tumors for deep imaging, enables superior photoacoustic contrast. Glypican-3 (GPC3) specific peptide (Kd = 71 nM) conjugated gold nanoshell (λabs=770nm) was used to detect HCC xenograft tumors in mice with photoacoustic imaging. This targeting probe demonstrated tumor uptake after 1 hr and cleared in 12 hrs. Images at a mean (±SD) depth of 9.7±1.4 mm from 0 to 2.1 cm beneath the skin revealed increased PA signal from tumors. Highest tumor uptake and tumor to normal tissue ratio occurred at 2 hrs post injection (T/B = 4.45±0.22, n = 8). Molecular targeting GNS showed potential as a simple, effective and rapid technique for noninvasive in vivo monitoring HCC tumor growth and GPC3 expression.
Aimed to build a dual-axes confocal endomicroscope with an outer diameter of 5.5mm for in-vivo imaging applications, an electrostatic MEMS scanner has been developed to enable two dimensional (2D) light scanning in either horizontal plane or vertical cross-sectional plane. The device has a compact structure design to match the dual axes confocal architecture in the probe without blocking the collimated light beams of excitation and collection, and a cutting-free silicon-on-insulator(SOI) micromachining process is used for the fabrication. A novel lever-based gimbal-like mechanism is employed to enable three degrees of freedom motions for lateral and axial light scanning, and its geometry is optimized for achieving large deflection with high scanning speed. Based on parametric excitation, the device can work in resonant modes. Testing result shows that, up to ±27° optical deflection angle for inner axis torsion motion with a frequency of ~4.9kHz, up to ±28.5° optical deflection angle for outer axis torsion motion with a frequency of~0.65kHz and ~360μm stroke for out-of-plane translation motion with a frequency of ~0.53kHz are achieved with <60V driving voltage. Based on these results, 2D imaging with frame rate of 5~10Hz and large field of view (1000μm x 1000μm in horizontal plane and 1000μm x 400μm in vertical plane) can be enabled by this scanner.
Tremendous advances have been made in technological development of whole body molecular imaging, including PET, SPECT, MRI, bioluminescence, and ultrasound. However, a great unmet need still exists for high resolution imaging of biological processes that occur in the epithelium, the thin layer of tissue where many important cancers originate. Confocal endomicroscopes designed with a fiber bundle are used in the clinic, but they can only create images in the horizontal plane. Imaging in the plane perpendicular to the tissue surface is also important because epithelial cells differentiate in the vertical direction. Subtle changes in normal tissue differentiation patterns can reveal the early expression of cancer biomarkers.
In this work, we present a side-viewing confocal endomicroscope that can collect images in either horizontal or oblique plane using an integrated monolithic electrostatic 3D MEMS scanner. The endomicroscope can perform sub-cellular resolution imaging in both the horizontal plane and the oblique plane with FOVs of 500 x 700 µm2 and 500 x 200 µm2. A side-viewing probe will allow optimal contact between the imaging window and the luminal wall, which makes it easy to navigate in the hollow organ. The endomicroscope is packaged into a stainless steel tube with outer diameter of 4.2 mm, which can be used for both small animal and human GI tract imaging. We demonstrate in vivo imaging of colonic dysplasia in mice, showing the endomicroscope can potentially be used for early detection and staging of colon cancer.
We demonstrate a dual axes confocal architecture, which can be used to collect horizontal(XY-plane) or vertical cross-sectional(XZ-plane) images for tissue. This scanner head is 5.5mm in outer diameter(OD), and integrates a 3D MEMS scanner with a compact chip size of 3.2×2.9mm2. To realize the miniaturization, there are some obstacles of the small size of 3D MEMS scanner, MEMS wire bundle, the air pressure effect for MEMS motion, the processing of parabolic mirror, and optical alignment to come over. In our probe, separation mechanical structure for optical alignment was adopted and a step shape MEMS holder was designed to deal with the difficult of MEMS wire bundle. Peptides have been demonstrated tremendous potential for in vivo use to detect colonic dysplasia. This class of in vivo molecular probe can be labeled with near-infrared (NIR) dyes for visualizing the full depth of the epithelium in small animals. To confirm our probe performance, we take use of USAF 1951 resolution target to test its lateral and axial resolution. It has lateral and axial resolution of 2.49um and 4.98um, respectively. When we collect the fluorescence imaging of colon, it shows that the field of view are 1000um×1000um (horizontal) and 1000um×430um (vertical). The horizontal and vertical cross-sectional images of fresh mouse colonic mucosa demonstrate imaging performance with this miniature instrument.
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