We describe a substantially improved NIRF-IVUS imaging system to overcome current technical limitations. We have implemented a hybrid rotary joint capable of rotational speeds up to 6,000rpm and developed a NIRF-IVUS imaging catheter with a robust dual-layer drive shaft and a reduced rigid length of 2mm with a catheter size <3.6F. NIRF-IVUS processing software was also improved by implementing a high-speed acquisition trigger and data streaming for fast recording speeds. NIRF-IVUS imaging at speeds of at least 30 fps in phantoms and in vivo arterial disease models will demonstrate the unique capabilities of IVUS-NIRF imaging of plaque pathobiology.
We conducted a first-in-human study using a fluorescent slurry comprised of 10 ml Methylene Blue (50 mg dissolved in water) and 10 ml Agave nectar. Preliminary data from the first patient indicates that at 20 minutes Barrett's Esophagus (esophageal pre-cancer) can be discriminated from normal tissue with a target-to-background ratio of 1.96 and an effect size > 5 (Cohen's D). We further show a dose modification pathway to high-TBR imaging shortly after swallowing the slurry.
Colorectal cancer (CRC) is one of the top causes of malignancy in both men and women. Although screening has significantly reduced CRC mortality, colonoscopy suffers from inadequate inspection and sampling of the tissue, a limitation that could be addressed by Optical Coherence Tomography (OCT). However, thus far, most studies have concentrated on the qualitative evaluation of morphological features and, only recently, the automatic classification of OCT images is being explored. To improve the classification of human tissues, manual or automatic, the spectral information in the OCT interferogram can be exploited. It can provide additional information regarding disease-related absorption and/or scattering changes in the tissue. In this study, we propose the use of multi-spectral analysis of OCT images, i.e. the utilization of images created from different bands of the available spectrum, to classify human colon polyps as normal or abnormal. Multiple, narrow-band, images, at different center wavelengths, were combined to create a “spectral score” for each pixel of the image. This fusion of information allowed both easier visual evaluation of the images as well as automatic classification (80 % accuracy per patient with leave-one-patient-out cross-validation). The proposed approach must be expanded to include more polyps and explore more sophisticated multi-spectral deep learning methods to improve its accuracy. However, these preliminary results provide evidence that this method has the potential to improve the accuracy of OCT and, in the future, enable clinical applications for colon cancer diagnosis.
OCT-based tethered capsule endomicroscopy (TCE) is an emerging tool for unsedated Barrett’s Esophagus (BE) screening. Cancer progression risk is best determined by acquiring and analyzing a BE tissue sample. We report a TCE device with a biopsy channel capable of extracting tissue samples in an unsedated platform. We show in swine studies (n=2) the biopsy capsule can locate simulated targets and visualize the extraction of biopsies. Owing to its capacity to be utilized in patients without requiring sedation, this new technology could be useful for screening for BE subjects who have an elevated risk of developing cancer.
We have developed a new self-propelled OCT imaging technology called retrograde Tethered Capsule Endomicroscopy
(R-TCE) for colonic disease screening. We successfully demonstrated that the R-TCE device can be advanced over 1 meter in 5 swine colons in vivo. R-TCE with balloon pullback imaging enabled full circumferential OCT visualization of 95.94 % ± 0.13% of the colon wall. 3D reconstructed colon OCT images and 3D rendered flythroughs showed that R-TCE is feasible for OCT microscopic imaging of the entire colon in vivo. When translated to humans, this R-TCE
technology may provide a less invasive and more efficient alternative to colonoscopy.
Colorectal cancer (CRC) is one of the top causes of malignancy in both men and women. Although screening has significantly reduced CRC mortality, colonoscopy suffers from inadequate inspection and sampling of the tissue, a limitation that can be addressed by Optical Coherence Tomography (OCT). The use of fractal analysis, which has been shown to estimate the scatterer size from OCT data, could help improve the classification of colon polyps as compared to using morphological information alone. For this study, thirty polyps were imaged immediately post excision, histologically processed, and both the OCT and H&E images annotated by an expert. Multi-step segmentation was used to segment the crypt regions. Fractal analysis of those areas was employed to estimate the nuclear size and classify the polyps as normal or tubular adenoma. This process resulted in an accuracy of 81% (92% sensitivity, 40% specificity) as confirmed by histology. The poor specificity can be partially attributed to the small number (only 5) of normal polyps and, more importantly, to the confirmed presence of other histological features which might influence the fractal analysis. The proposed approach must be expanded to include more polyps and further enhanced to improve its specificity. However, these preliminary results provide evidence that this method has the potential to perform scatterer size estimation and tissue classification from en face images, thus, providing a robust approach for the improvement of the accuracy of endoscopic OCT and, in the future, the effectiveness of colonoscopy.
We report the use of our multimodal near-infrared fluorescence (NIRF) and OCT imaging system and catheter to perform the first imaging of LUM015 inflammatory activity in rabbit models of atherosclerosis in vivo. Using co-injection and multi-channel intravascular NIRF-OCT, we compared LUM015 (6.2 mg/kg) and preclinical ProSense (VM110, 3.5 mg/kg) fluorescence in the same subject. We found that co-registered fluorescence carpet maps were remarkably similar with a PCC of 0.51 and a Mander’s overlap coefficient of 0.79. Results suggest that LUM015 will be a viable clinical option for intracoronary imaging of plaque inflammatory activity in patients.
Significance: Confocal laser scanning enables optical sectioning in fiber bundle endomicroscopy but limits the frame rate. To be able to better explore tissue morphology, it is useful to stitch sequentially acquired frames into a mosaic. However, low frame rates limit the maximum probe translation speed. Line-scanning (LS) confocal endomicroscopy provides higher frame rates, but residual out-of-focus light degrades images. Subtraction-based approaches can suppress this residue at the expense of introducing motion artifacts.
Aim: To generate high-frame-rate endomicroscopy images with improved optical sectioning, we develop a high-speed subtraction method that only requires the acquisition of a single camera frame.
Approach: The rolling shutter of a CMOS camera acts as both the aligned and offset detector slits required for subtraction-based sectioning enhancement. Two images of the bundle are formed on different regions of the camera, allowing both images to be acquired simultaneously.
Results: We confirm improved optical sectioning compared to conventional LS, particularly far from focus, and show that motion artifacts are not introduced. We demonstrate high-speed mosaicing at frame rates of up to 240 Hz.
Conclusion: High-speed acquisition of optically sectioned images using the new subtraction based-approach leads to improved mosaicing at high frame rates.
Significance: Confocal laser scanning enables optical sectioning in clinical fiber bundle endomicroscopes, but lower-cost, simplified endomicroscopes use widefield incoherent illumination instead. Optical sectioning can be introduced in these simple systems using structured illumination microscopy (SIM), a multiframe digital subtraction process. However, SIM results in artifacts when the probe is in motion, making the technique difficult to use in vivo and preventing the use of mosaicking to synthesize a larger effective field of view (FOV).
Aim: We report and validate an automatic motion compensation technique to overcome motion artifacts and allow generation of mosaics in SIM endomicroscopy.
Approach: Motion compensation is achieved using image registration and real-time pattern orientation correction via a digital micromirror device. We quantify the similarity of moving probe reconstructions to those acquired with a stationary probe using the relative mean of the absolute differences (MAD). We further demonstrate mosaicking with a moving probe in mechanical and freehand operation.
Results: Reconstructed SIM images show an improvement in the MAD from 0.85 to 0.13 for lens paper and from 0.27 to 0.12 for bovine tissue. Mosaics also show vastly reduced artifacts.
Conclusion: The reduction in motion artifacts in individual SIM reconstructions leads to mosaics that more faithfully represent the morphology of tissue, giving clinicians a larger effective FOV than the probe itself can provide.
We report a compact rigid instrument capable of delivering en-face optical coherence tomography (OCT) images alongside (epi)-fluorescence endomicroscopy (FEM) images by means of a robotic scanning device. Two working imaging channels are included: one for a one-dimensional scanning, forward-viewing OCT probe and another for a fiber bundle used for the FEM system. The robotic scanning system provides the second axis of scanning for the OCT channel while allowing the field of view (FoV) of the FEM channel to be increased by mosaicking. The OCT channel has resolutions of 25 / 60 μm (axial/lateral) and can provide en-face images with an FoV of 1.6 × 2.7 mm2. The FEM channel has a lateral resolution of better than 8 μm and can generate an FoV of 0.53 × 3.25 mm2 through mosaicking. The reproducibility of the scanning was determined using phantoms to be better than the lateral resolution of the OCT channel. Combined OCT and FEM imaging were validated with ex-vivo ovine and porcine tissues, with the instrument mounted on an arm to ensure constant contact of the probe with the tissue. The OCT imaging system alone was validated for in-vivo human dermal imaging with the handheld instrument. In both cases, the instrument was capable of resolving fine features such as the sweat glands in human dermal tissue and the alveoli in porcine lung tissue.
Endomicroscopy is a technique for obtaining real-time images in vivo, eliminating the need to biopsy a tissue sample. A simple fluorescence endomicroscope can be constructed using a fiber bundle, camera, LED and filters, and individual images can be mosaicked as the probe is moved across the tissue to increase the image size. However, to improve image contrast optical sectioning is required for the removal of returning out-of-focus light. Commonly, this is done using the confocal technique, requiring more expensive laser sources and mechanical scanning mirrors which limits the frame rate. Structured illumination microscopy (SIM) instead uses line patterns projected onto the sample to allow for computational optical sectioning. This eliminates the need for point scanning and allows an incoherent light source, such as an LED, to be used, at the cost of some loss of signal-to-noise ratio. However, as SIM requires multiple images to be combined, motion of the probe results in severe image artefacts, preventing the use of mosaicking techniques. We report a SIM endomicroscope using a digital micro-mirror device (DMD) to generate line patterns at high speed, and with the ability to change the patterns on the fly. Combined with a high-speed camera, this reduces motion artefacts significantly, but not sufficiently to allow for video mosaicking techniques. We therefore demonstrate further reduction of artefacts by orienting the illumination patterns parallel to the direction of motion and performing inter-frame registration and correction. This offers potential for low cost, versatile, optically-sectioned endomicroscopy.
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