Hanh N. D. Le

Multispectral tissue characterization for intestinal anastomosis optimization

Abstract. Intestinal anastomosis is a surgical procedure that restores bowel continuity after surgical resection to treat intestinal malignancy, inflammation, or obstruction. Despite the routine nature of intestinal anastomosis procedures, the rate of complications is high. Standard visual inspection cannot distinguish the tissue subsurface and small changes in spectral characteristics of the tissue, so existing tissue anastomosis techniques that rely on human vision to guide suturing could lead to problems such as bleeding and leakage from suturing sites. We present a proof-of-concept study using a portable multispectral imaging (MSI) platform for tissue characterization and preoperative surgical planning in intestinal anastomosis. The platform is composed of a fiber ring light-guided MSI system coupled with polarizers and image analysis software. The system is tested on ex vivo porcine intestine tissue, and we demonstrate the feasibility of identifying optimal regions for suture placement.

An average enumeration method of hyperspectral imaging data for quantitative evaluation of medical device surface contamination

We propose a quantification method called Mapped Average Principal component analysis Score (MAPS) to enumerate the contamination coverage on common medical device surfaces. The method was adapted from conventional Principal Component Analysis (PCA) on non-overlapped regions of a full frame hyperspectral image to resolve the percentage of contamination from the substrate. The concept was proven by using a controlled contamination sample with artificial test soil and color simulating organic mixture, and was further validated using a bacterial system including biofilm on stainless steel surface. We also validate the results of MAPS with other statistical spectral analysis including Spectral Angle Mapper (SAM). The proposed method provides an alternative quantification method for hyperspectral imaging data, which can be easily implemented by basic PCA analysis.

Three-dimensional facial digitization using advanced digital image correlation

Presented in this paper is an effective technique to acquire the three-dimensional (3D) digital images of the human face without the use of active lighting and artificial patterns. The technique is based on binocular stereo imaging and digital image correlation, and it includes two key steps: camera calibration and image matching. The camera calibration involves a pinhole model and a bundle-adjustment approach, and the governing equations of the 3D digitization process are described. For reliable pixel-to-pixel image matching, the skin pores and freckles or lentigines on the human face serve as the required pattern features to facilitate the process. It employs feature-matching-based initial guess, multiple subsets, iterative optimization algorithm, and reliability-guided computation path to achieve fast and accurate image matching. Experiments have been conducted to demonstrate the validity of the proposed technique. The simplicity of the approach and the affordable cost of the implementation show its practicability in scientific and engineering applications.

Through-skull vasculature assessment using fluorescence brain imaging on murine models at around 800 nm

We describe a scanning near-infrared fluorescence imager for through-skull non-invasive brain imaging on live murine models. The captured photoluminescence feature through scattering media was enhanced using a high sensitivity scientific CMOS sensor with the obtained spatial resolution of 15.63 μm, depth of field of 5 mm and an average local signal-to-noise ratio of 37.5 dB.

Recording membrane potential changes through photoacoustic voltage sensitive dye

Monitoring of the membrane potential is possible using voltage sensitive dyes (VSD), where fluorescence intensity changes in response to neuronal electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. In contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near infrared light excitation and ultrasound detection. In this work, we develop the theoretical concept whereby the voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. Based on this concept, we synthesized a novel near infrared photoacoustic VSD (PA-VSD) whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. With a 3-9 μM VSD concentration, we measured a PA signal increase in the range of 5.3 % to 18.1 %, and observed a corresponding signal reduction in fluorescence emission of 30.0 % to 48.7 %. A theoretical model successfully accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate the voltage sensing capability of the dye, but also indicate the necessity of considering both fluorescence and absorbance spectral sensitivities in order to optimize the characteristics of improved photoacoustic probes. Together, our results demonstrate photoacoustic sensing as a potential new modality for sub-second recording and external imaging of electrophysiological and neurochemical events in the brain.

Listening to membrane potential: photoacoustic voltage-sensitive dye recording

Abstract. Voltage-sensitive dyes (VSDs) are designed to monitor membrane potential by detecting fluorescence changes in response to neuronal or muscle electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. By contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near-infrared light excitation and ultrasound detection. Here, we show that voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. We synthesized a near-infrared photoacoustic VSD (PA-VSD), whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. A theoretical model accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate PA voltage sensing but also emphasize the interplay of both fluorescence and absorbance properties in the design of optimized PA probes. Together, our results demonstrate PA sensing as a potential new modality for recording and external imaging of electrophysiological and neurochemical events in the brain.

Monitoring biofilm attachment on medical devices surfaces using hyperspectral imaging

Microbial biofilm is a colony of single bacteria cells (planktonic) that attached to surfaces, attract other microorganisms to attach and grow, and together they build an extracellular matrix composed of polysaccharides, protein, and DNA. Eventually, some cells will detach and spread to other surface. Biofilm on medical devices can cause severe infection to all age ranges from infant to adult. Therefore, it is important to detect biofilm in a fast and efficient manner. Hyperspectral imaging was utilized for distinguishing wide area of biofilm coverage on various materials and on different textures of stainless steeltest coupons. Not only is the coverage of biofilm important, but also the shear stress of biofilm on the attached surfaces is significant. This study investigates the effects of shear stress on the adhesion of biofilms on common medical device surfaces such as glass, polycarbonate, polytetrafluoroethylene, and stainless steel with different textures. Biofilm was grown using Ps. aeruginosa and growth was monitored after 24 and 48 hours at 37° C. The coupons covered with biofilm were tilted at 45 degrees and 90 degrees for 30 seconds to induce shear stress and Hyperspectral images were taken. We hypothesize that stronger attachment on rough surface would be able to withstand greater shear stress compared to smooth surface.

An endoscopic 3D structured illumination imaging system for robotic anastomosis surgery (Conference Presentation)

Surgical 3D endoscopy based on structured illumination has been built and evaluated for application in minimally invasive anastomosis surgery which offers advantages of smaller incision, low risk of infection, quick recovery times and reduced blood loss. When combined with robotic manipulations, surgeons can perform surgical tasks with higher precision and repeatability. For reconstructive surgery such as anastomosis, a supervised laparoscopic anastomosis using a surgical robot has recently been reported with an open-surgery approach using a large 3D camera. To push the technology into minimally-invasive setting, we report an endoscopic 3D system based on structured illumination technique to assist the surgical robot, particularly in anastomosis surgery. The recorded structural profile achieves a high depth quantification of 250 um for static objects, with 25 mm depth of field. The proposed system can be integrated into a flexible holding arm to move in accordance with the surgical robotic arm. We characterize the system performance using multiple porcine intestinal tissue samples with variations in surface textures, tissue pigmentation and thickness.

Real-time intra-operative guidance using combined photoacoustic and pulsed fluorescence imaging for robot-assisted surgical operation

Multi-modal interface in medicine has been of interest as it extends clinical vision and diagnostic to a higher dimension. In this paper, we develop a real-time intra-operative guidance for robot-assisted surgical operation using a dual photoacoustic (PA) and fluorescence (FL) imaging based on a common pulsed laser and commercial ultrasound array transducer.

3D endoscopic imaging using structured illumination technique (Conference Presentation)

Surgeons have been increasingly relying on minimally invasive surgical guidance techniques not only to reduce surgical trauma but also to achieve accurate and objective surgical risk evaluations. A typical minimally invasive surgical guidance system provides visual assistance in two-dimensional anatomy and pathology of internal organ within a limited field of view. In this work, we propose and implement a structure illumination endoscope to provide a simple, inexpensive 3D endoscopic imaging to conduct high resolution 3D imagery for use in surgical guidance system. The system is calibrated and validated for quantitative depth measurement in both calibrated target and human subject. The system exhibits a depth of field of 20 mm, depth resolution of 0.2mm and a relative accuracy of 0.1%. The demonstrated setup affirms the feasibility of using the structured illumination endoscope for depth quantization and assisting medical diagnostic assessments