Although picrosirius red (PSR) is known to be in quantifying collagen under polarized light (PL), commonly used linearly PL can result in an underestimation of collagen, as some of the fibers may appear dark if aligned with the transmission axis of the polarizers. To address this, a sample may be imaged with circularly polarized light at the expense of higher background intensity. However, the quality and alignment of the microscope illumination optics, polarizers and waveplates can still produce imaging variability with circular polarization. A simpler technique was tested that minimized variability and background intensity with linear polarization by acquiring images at multiple angles of histology slide rotation to create a composite co-registered image, permitting the optimal semi-quantitative visualization of collagen.
Linear polarization imaging was performed on PSR stained artery sections. By rotating the slide at 60° intervals while maintaining illumination, polarization and exposure parameters, 6 images were acquired for each section. A composite image was created from the 6 co-registered images, and comprised of the maximum pixel intensity at each point.
Images from any of the 6 rotation positions consistently showed variation in PSR signal. A composite image compensates for this variability, without loss of spatial resolution. Additionally, grayscale analysis showed an increased intensity range of 15 – 50% with a linearly polarized composite image over a circularly polarized image after background correction, indicating better SNR. This proposed technique will be applied in the development of a near infrared spectroscopy algorithm to detect vulnerable atherosclerotic plaques in vivo.
The ability to determine plaque cap thickness during catheterization is thought to be of clinical importance for plaque vulnerability assessment. While methods to compositionally assess cap integrity are in development, a method utilizing currently available tools to measure cap thickness is highly desirable. NIRS-IVUS is a commercially available dual imaging method in current clinical use that may provide cap thickness information to the skilled reader; however, this is as yet unproven.
Ten autopsy hearts (n=15 arterial segments) were scanned with the multimodality NIRS-IVUS catheter (TVC Imaging System, Infraredx, Inc.) to identify lipid core plaques (LCPs). Skilled readers made predictions of cap thickness over regions of chemogram LCP, using NIRS-IVUS. Artery segments were perfusion fixed and cut into 2 mm serial blocks. Thin sections stained with Movat’s pentachrome were analyzed for cap thickness at LCP regions. Block level predictions were compared to histology, as classified by a blinded pathologist.
Within 15 arterial segments, 117 chemogram blocks were found by NIRS to contain LCP. Utilizing NIRSIVUS, chemogram blocks were divided into 4 categories: thin capped fibroatheromas (TCFA), thick capped fibroatheromas (ThCFA), pathological intimal thickening (PIT)/lipid pool (no defined cap), and calcified/unable to determine cap thickness. Sensitivities/specificities for thin cap fibroatheromas, thick cap fibroatheromas, and PIT/lipid pools were 0.54/0.99, 0.68/0.88, and 0.80/0.97, respectively. The overall accuracy rate was 70.1% (including 22 blocks unable to predict, p = 0.075). In the absence of calcium, NIRS-IVUS imaging provided predictions of cap thickness over LCP with moderate accuracy. The ability of this multimodality imaging method to identify vulnerable coronary plaques requires further assessment in both larger autopsy studies, and clinical studies in patients undergoing NIRS-IVUS imaging.
Intracoronary near-infrared spectroscopy (NIRS) can identify lipid in the coronary arteries, but lacks depth resolution. A
novel catheter is currently in clinical use that combines NIRS with intravascular ultrasound (IVUS), which provides
depth-resolved structural information via the IVUS modality. A measure designated as lipid-rich plaque burden (LRPB)
has been proposed as a means to interpret the combined acoustic and optical information of NIRS-IVUS. LRPB is
defined as the area created by the intersection of the NIRS lipid-rich arc with the corresponding IVUS-measured plaque
burden. We determined the correlation in human coronary autopsy specimens between LRPB, a measure of lipid
presence and extent available via intravascular imaging in patients, and the area of lipid-rich plaque as determined by the
gold-standard of histology.
Fifteen artery segments from 8 human autopsy hearts were imaged with the NIRS-IVUS system (TVC Imaging System,
Infraredx Inc., Burlington, MA). Arteries were imaged in a specialty fixture that assured accurate co-registration
between imaging and histology. The arteries were then fixed and divided into 2 mm blocks for histological staining.
Pathological contouring of lipid-rich areas was performed on the stained thin sections for 54 lipid-rich blocks.
Computation of LRPB was performed on transverse NIRS-IVUS frames corresponding to the histologic sections. The
quantified LRPB was frequently higher than the lipid-rich plaque area determined by histology, because the region
denoted by the EEL and lumen within the NIRS lipid-rich arc is not entirely comprised of lipid. Overall, a moderate to
strong correlation (R = 0.73) was found between LRPB determined by NIRS-IVUS imaging and the lipid-rich plaque
area determined by histology. LRPB, which can be measured in patients with NIRS-IVUS imaging, corresponds to the
amount of lipid-rich plaque in a coronary artery. LRPB should be evaluated in prospective clinical trials for its ability to
identify vulnerable plaques.
Metal needles are commonly used for drug delivery or biopsy collection in clinical settings. Needle deflection and
deformation can occur when inserting needles into soft,
non-homogeneous tissues which can affect the location accuracy
of insertion. Therefore, the ability to visualize both anatomical surrounding structures and the advancing needle is
required. Ultrasound is commonly used for image-guidance of needles; however, specular reflections from the metal
surface can deflect the acoustic beam away from the transducer when the needle is even slightly angled from the US
transducer thereby rendering the needle invisible in the image.
Photoacoustic imaging has been proposed for guidance of metal needles and other metal objects in-vivo. The high
optical absorption coefficient of stainless steel can provide high photoacoustic imaging contrast. The photoacoustic
signal is produced omni-directionally from the metal surface allowing for greater detection of needles at increasing
injection angles compared to ultrasound imaging. In the current work, needles were inserted into excised tissue and
imaged using an ultrasound array transducer and a pulsed 800 nm laser. The results showed that at a shallow 10°
insertion angle, the photoacoustic ratio of needle signal to background was four-times higher compared to ultrasound.
Furthermore, the surrounding tissue composition was observed to have an effect on photoacoustic signal enhancement
which correlated with the change of the Grüneisen coefficient of the surrounding tissue environment, suggesting that the
photoacoustic signal amplitude could be used to ascertain surrounding tissue composition. Photoacoustic imaging
provides sufficient depth penetration for this application and offers excellent image contrast.
The ability to visualize and track temporarily or permanently implanted metal devices is important in many applications ranging from diagnosis to therapy. Specifically, reliable imaging of metal needles is required in today's clinical settings. Currently, ultrasound is utilized to image a needle inserted into tissue in real time. However, the diagnostic value and tracking ability of these images depends highly on the orientation of the needle, and also its proximity to regions of interest in the tissue. We examine the use of photoacoustic imaging combined with current ultrasound imaging methods to obtain high-contrast images of commonly used needles in the body. Experiments were performed using 21 G and 30 G needles inserted into ex vivo porcine tissue and tissue-mimicking phantoms. The needles and surrounding tissue were imaged using an ultrasound imaging system interfaced with the pulsed laser source necessary for photoacoustic imaging. The results suggest that photoacoustic imaging, combined with ultrasound imaging, is capable of real-time, high-contrast, and high-spatial-resolution visualization of metal implants within anatomical landmarks of the background tissue.
An acceleration of angiogenesis in the adventitial vasa-vasorum is usually associated with vulnerable, thin-cap fibroatheroma
in atherosclerotic plaques. Angiogenesis creates microvasculature too small to be detected and differentiated
using conventional imaging techniques. However, by using spectroscopic photoacoustic imaging, we take advantage of
the wavelength-dependent optical absorption properties of blood. We used a vessel-mimicking phantom with micro
blood vessels. The phantom was imaged with intravascular photoacoustic imaging across a range of wavelengths. The
image intensities were cross-correlated with the known absorption spectra of blood. The resulting cross-correlation
image was able to reveal the location of the artificial blood vessels differentiated from non-blood vessel components.
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