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Since the nationwide launch of the Standardized Residency Training Program (SRTP) in China, residents have faced multiple barriers from work and the environment, and their empathy, work-related perception, and clinical performance are constantly scrutinized by training administrators and educators. This study aimed to investigate the influence of empathy of residents on their clinical performance and the possible mediating role of work-related perception. A multi-center cross-sectional study was conducted including 1160 residents from 10 cities in Liaoning Province between March and October 2020. Participants completed an array of self-reported questionnaires, regarding demographic characteristics, work-related perception, clinical performance, and empathy (measured by the Perspective Taking and Empathic Concern subscales of the Chinese version of Interpersonal Reactivity Index). The relationships among empathy, work-related perception, and clinical performance were examined using a structural equation model (SEM). Of the 1160 residents, 961 (82.8%) completed the questionnaires. The final SEM produced a good fit to the sample data, with CMIN/DF = 2.11 (P

The distributed temperature sensing (DTS) is used to overcome the defects of traditional production profile testing technology and realize the real-time and accurate temperature monitoring of complex underground reservoirs. However, the temperature response characteristics of the production profile for multilayered gas wells have not been studied clearly, which leads to technical problems of the production profile interpretation of such gas wells monitored by DTS. Therefore, considering the influence of the fluid heat convection, viscous dissipation, and heat conduction which are the involved mechanisms in this process, a model coupling pressure and temperature fields of a multilayered gas reservoir are established in the current study. Subsequently, the formation temperature variation for transient testing of a multilayered gas reservoir is solved by programming, and the effect of model parameters (e.g., gas production, permeability, and rock heat capacity ratio) on the temperature response characteristics of the production profile is analyzed. Finally, the accuracy and reliability of temperature response prediction are verified by fitting the actual DTS temperature test data of an offshore-multilayered gas well. The results of this study provide ideas regarding quantitative interpretation and analysis of DTS monitoring data for the production profile of multilayered gas wells.

Point-of-care medical diagnosis demands immediate feedback on tissue pathology. Confocal endomicroscopy can provide real-time in vivo images with histology-like features. The working channel in medical endoscopes are becoming smaller in dimension. Microsystems methods can produce tiny mechanical scanners. We demonstrate a flexible fiber instrument for in vivo imaging as an endoscope accessory. The optical path is folded on-axis to reduce length while allowing the beam to expand and achieve a numerical aperture of 0.41. A high-speed parametric resonance mirror produces large deflection angles > 13°, and is mounted on a 2 mm diameter chip designed with clamp structures for reduced space. A compact lens assembly provides diffraction-limited lateral and axial resolution of 1.5 and $12~\mu \text{m}$ , respectively. A working distance of $50~\mu \text{m}$ and field-of-view of $350\,\,\mu \text{m}\,\,\times 350\,\,\mu $ m are achieved. Miniature apparatus is fabricated to assemble and align the scanhead components. The optics and scanner are packaged in a distal tip with 2.4 mm diameter and 10 mm rigid length. These dimensions allow the endomicroscope to pass forward easily through the 2.8 mm diameter working channel in medical endoscopes commonly used in clinical practice. Fluorescence images are collected in vivo at 10 frames per second in the colon of genetically-engineered mice that spontaneously develop adenomas. A FITC-labeled peptide heterodimer is administered intravenously to provide specific contrast. Sub-cellular structures are visualized to distinguish pre-malignant from normal mucosa. These results demonstrate use of microsystems methods to produce an ultra-compact instrument with sufficiently small dimensions for broad use.

BACKGROUND The management of people living with HIV/AIDS (PLWHA) is multidimensional and complex. It is increasingly recognized that the use of patient reported outcome measures (PROMs) is the key to providing patient-centered health care to meet the lifelong needs of PLWHA from diagnosis to the end of life. However, there is no consensus on which PROM should be recommended for healthcare providers and researchers to assess health outcomes of PLWHA currently. OBJECTIVE This review aims to assess HIV-specific PROMs for adult PLWHA using COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) methodology. METHODS This systematic review was guided by the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement. The literature search was conducted in 3 recommended databases (PubMed, Embase, and PsychINFO) on January 15, 2021. Eligible studies were included if they assessed any psychometric property of HIV-specific PROMs in adult PLWHA. The PROMs were assessed for 9 psychometric properties, which were assessed in each included study using the COSMIN methodology by assessing (1) the methodological quality assessed using the COSMIN risk of bias checklist, (2) overall rating of results, (3) level of evidence assessed using the modified Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach, and (4) level of recommendation. RESULTS A total of 88 PROMs in 8 categories were identified, assessing the psychometric properties of PROMs for adult PLWHA in 152 studies involving 79213 PLWHA. The psychometric properties of the majority of included PROMs were rated with insufficient evidence. The PROMs that received class A recommendation was the Poz Quality of Life, HIV Symptom Index or Symptoms Distress Module of the ACTG, and People Living with HIV Resilience Scale. In addition, due to insufficient evidence, most of remaining assessed PROMs cannot make recommendations regarding their use (received class B recommendation). CONCLUSIONS This systematic review recommended 3 PROMs to assess health outcomes of adult PLWHA, however, they all have some shortcomings. It is necessary to conduct a more thorough assessment of the psychometric properties of existing PROMs. These findings may provide a reference to the selection of high-quality HIV-specific PROMs by healthcare providers and researchers for clinical practice and research.

Pentostatin is a nucleoside antibiotics with a strong inhibitory effect on adenosine deaminase, and is widely used in the clinical treatment of malignant tumors. However, the high cost hampers its application. In the past 10 years, the biosynthesis of pentostatin were focused on strain breeding, optimization of medium composition and fermentation process. To date, there are no reviews summarizing the elucidated biosynthetic mechanism of pentostatin. This review starts by introducing the various chemical route for production of pentostatin, followed by summarizing the mechanisms of pentostatin biosynthesis in different microorganisms. Finally, challenges for biosynthesis of pentostatin were discussed, and strategies for regulating and improving the microbial synthesis of pentostatin were proposed.

Electrostatic microactuators with large vertical scanning range (several hundred micrometers) at high frequency (hundreds to thousands of hertz) and chips’ sizes compatible with endoscopic microscopy have recently been demonstrated based on parametric resonance. This paper examines the use and modeling of mixed softening/hardening dynamics to help produce large ranges of motion in this class of mirrors. Origin of spring stiffening behavior in actuator design is described, followed by non-dimensional analysis of actuator motion trends. The experimental results are presented for a sample actuator design with up to 480- $\mu \text{m}$ displacement at 1225 Hz and 60 V. Comparison of predicted trends and comments on benefits and limitations of modeling are provided. [2018-0184]

This paper presents an optical lens design for laser projector induced speckle measurement, based on resolution cells of human eyes. While the speckle pattern at the image plane was split up into hundreds of cells of human eyes resolution and formed cells pattern, its contrast become lower than that was calculated based on individual pixels. To evaluate the sensation of human eyes on the speckle pattern and perform a standardized speckle measurement, resolution of human eyes at the objective plane should be taken into consideration. Different lenses of a camera sensor usually have different field of views, therefore each pixel presents different size at the objective plane. The relationship between pixel size on the objective plane, human eyes resolution and F-number of camera lens was deduced, which is important for the optical design for speckle measurement. An optical design around an Aptina’s MT9P001 imaging sensor was developed, optimized and analyzed using Zemax. The results show that the MTF is above 0.3 when spatial frequency is 113.5 lp/mm, distortion is less than 1%, relative illumination is more than 90% at the max field of view, and the radius of the spot size is less than 2um, which has met the requirements for speckle pattern measurement based on human eyes resolution cell. Finally, we performed a sensitivity analysis of tolerance and Monto Carlo simulation to confirm that the lens could be fabricated.

This paper applies image processing metrics to tracking of perturbations in mechanical phase delay in a multi-axis microelectromechanical system (MEMS) scanner. The compact mirror is designed to scan a laser beam in a Lissajous pattern during the collection of endoscopic confocal fluorescence images, but environmental perturbations to the mirror dynamics can lead to image registration errors and blurry images. A binarized, threshold-based blur metric and variance-based sharpness metric are introduced for detecting scanner phase delay. Accuracy of local optima of the metric for identification of phase delay is examined, and relative advantages for processing accuracy and computational complexity are assessed. Image reconstruction is demonstrated using both generic images and sample tissue images, with significant improvement in image quality for tissue imaging. Implications of non-ideal Lissajous scan effects on phase detection and image reconstruction are discussed.

During operation in endoscopic imaging instruments, on-chip motion sensing by microelectromechanical system (MEMS) scanning mirrors can be difficult to perform due to high scanning frequencies and poorly known nonlinear behavior in system dynamics. In this work, we optimize a nonlinear model for on-chip capacitance simultaneously with a simple neural net model for unknown transmission line and circuit dynamics. Use of nonlinear model elements is observed to improve accuracy of predicted sensor output during ringdown, parasitic feedthrough, and multi-axis operation experiments. Calibration can be performed using a single iteration of ringdown and feed through training data. Motion estimation accuracy of approximately 0.1 rad of scan angle and 0.15 rad of phase delay is achieved, sufficient for diagnostic use in endoscopic imaging systems.

We present a method to estimate high frequency rotary motion of a highly compact electrostatic micro-scanner using the same electrodes for both actuation and sensing. The accuracy of estimated rotary motion is critical for reducing blur and distortion in image reconstruction applications with the micro-scanner given its changing dynamics due to perturbations such as temperature. To overcome the limitation that no dedicated sensing electrodes are available in the proposed applications due to size constraints, the method adopts electromechanical amplitude modulation (EAM) to separate motion signal from parasitic capacitance feedthrough, and a novel non-linear measurement model is derived to characterize the relationship between large out-of-plane angular motion and circuit output. To estimate motion, an extended Kalman filter (EKF) and an unscented Kalman filter (UKF) are implemented, incorporating a process model based on the micro-scanner’s parametric resonant dynamics and the measurement model. Experimental results show that compared to estimation without using the measurement model, our method is able to improve the rotary motion estimation accuracy of the micro-scanner significantly, with a reduction of root-mean-square error (RMSE) in phase shift of 86.1%, and a reduction of RMSE in angular position error of 78.5 %.

To explore the correlation between altered levels of neurotransmitters in the frontal lobe and hippocampus and behavioral abnormalities in a ClockOpen field test and Elevated plus-maze test were carried out on the ClockThe ClockClock

Esophageal adenocarcinoma (EAC) is a molecularly heterogeneous disease that is rising rapidly in incidence and has poor prognosis. We developed a heterobivalent peptide to target detection of early Barrett's neoplasia by combining monomer heptapeptides specific for either EGFR or ErbB2 in a heterodimer configuration. The structure of a triethylene glycol linker was optimized to maximize binding interactions to the surface receptors on cells. The Cy5.5-labeled heterodimer QRH*-KSP*-E3-Cy5.5 demonstrated specific binding to each target and showed 3-fold greater fluorescence intensity and 2-fold higher affinity compared with those of either monomer alone. Peak uptake in xenograft tumors was observed at 2 h postinjection with systemic clearance by ∼24 h in vivo. Furthermore, ligand binding was evaluated on human esophageal specimens ex vivo, and 88% sensitivity and 87% specificity were found for the detection of either high-grade dysplasia (HGD) or EAC. This peptide heterodimer shows promise for targeted detection of early Barrett's neoplasia in clinical study.

Design and testing of a low-mass electrothermal vertical scanning mirror for fast axial scanning during multi-photon microscopy is presented. The mirror makes use of a novel honeycomb mechanical support structure to substantially reduce both thermal and inertial mass relative to mirror size. At the same time, undesirable mirror curvature due to residual thermal stress is controlled without adversely affecting the image quality. In addition, the support structure makes the complete mirror area usable which otherwise is not possible in devices that rely on isotropic silicon etching. A high actuator efficiency with static displacement per unit power of 2.08 μm/mW was achieved due to the reduced thermal capacitance at the actuator legs, when using a 700 nm thin structural layer of SiO2. The resulting scanning mirror achieves open loop time constant as small as 3.42 ms with both thermal and mechanical bandwidths exceeding 100 Hz. Mirror surface non-idealities related to the mirror structure and their influence on image quality are discussed. Sample 3D multi-photon images captured by performing remote scanning in both lateral and axial direction on a benchtop testbed are presented. The efficacy of remote axial scan is ascertained by comparisons with images obtained by moving specimens relative to the objective.

The epithelium is a thin layer of tissue that lines hollow organs, such as colon. Visualizing in vertical cross sections with sub-cellular resolution is essential to understanding early disease mechanisms that progress naturally in the plane perpendicular to the tissue surface. The dual axes confocal architecture collects optical sections in tissue by directing light at an angle incident to the surface using separate illumination and collection beams to reduce effects of scattering, enhance dynamic range, and increase imaging depth. This configuration allows for images to be collected in the vertical as well as horizontal planes. We designed a fast, compact monolithic scanner based on the principle of parametric resonance. The mirrors were fabricated using microelectromechanical systems (MEMS) technology and were coated with aluminum to maximize near-infrared reflectivity. We achieved large axial displacements $> 400~\mu \text{m}$ and wide lateral deflections >20°. The MEMS chip has a $3.2\times2.9$ mm2 form factor that allows for efficient packaging in the distal end of an endomicroscope. Imaging can be performed in either the vertical or horizontal planes with $430~\mu \text{m}$ depth or $1 \times 1$ mm2 area, respectively, at 5 frames/s. We systemically administered a Cy5.5-labeled peptide that is specific for EGFR, and collected near-infrared fluorescence images ex vivo from pre-malignant mouse colonic epithelium to reveal the spatial distribution of this molecular target. Here, we demonstrate a novel scanning mechanism in a dual axes confocal endomicroscope that collects optical sections of near-infrared fluorescence in either vertical or horizontal planes to visualize molecular expression in the epithelium.

Objectives Conventional colonoscopy with white light illumination detects colonic adenomas based on structural changes alone and is limited by a high miss rate. We aim to demonstrate an integrated imaging strategy that combines wide-field endoscopy and confocal endomicroscopy in real time to visualize molecular expression patterns in vivo to detect premalignant colonic mucosa. Methods A peptide specific for claudin-1 is labeled with Cy5.5 and administrated intravenously in genetically engineered mice that develop adenomas spontaneously in the distal colon. Wide-field endoscopy is used to identify the presence of nonpolypoid and polypoid adenomas. Anatomic landmarks are used to guide placement of a confocal endomicroscope with side-view optics to visualize claudin-1 expression patterns with subcellular resolution. Results Wide-field fluorescence images show peak uptake in colon adenoma at ∼1 hour after systemic peptide administration, and lesion margins are clearly defined. Further examination of the lesion using a confocal endomicroscope shows dysplastic crypts with large size, elongated shape, distorted architecture, and variable dimension compared with normal. The mean fluorescence intensity is significantly higher for dysplasia than normal. Increased claudin-1 expression in dysplasia vs normal is confirmed ex vivo, and the binding pattern is consistent with the in vivo imaging results. Discussion Wide-field endoscopy can visualize molecular expression of claudin-1 in vivo to localize premalignant colonic mucosa, and confocal endomicroscopy can identify subcellular feature to distinguish dysplasia from normal.

Background & Aims: Conventional white-light colonoscopy aims to reduce the incidence and mortality of colorectal cancer (CRC). CRC has been found to arise from missed polypoid and flat precancerous lesions. We aimed to establish proof-of-concept for real-time endoscopic imaging of colonic adenomas using a near-infrared peptide that is specific for claudin-1. Methods: We used gene expression profiles to identify claudin-1 as a promising early CRC target, and performed phage display against the extracellular loop of claudin-1 (amino acids 53â80) to identify the peptide RTSPSSR. With a Cy5.5 label, we characterized binding parameters and showed specific binding to human CRC cells. We collected in vivo near-infrared fluorescence images endoscopically in the CPC;Apc mouse, which develops colonic adenomas spontaneously. With immunofluorescence, we validated specific peptide binding to adenomas from the proximal human colon. Results: We found a 2.5-fold increase in gene expression for claudin-1 in human colonic adenomas compared with normal. We showed specific binding of RTSPSSR to claudin-1 in knockdown and competition studies, and measured an affinity of 42 nmol/L and a time constant of 1.2 minutes to SW620 cells. In the mouse, we found a significantly higher target-to-background ratio for both polypoid and flat adenomas compared with normal by in vivo images. On immunofluorescence, we found significantly greater intensity for human adenomas (mean ± SD, 25.5 ± 14.0) vs normal (mean ± SD, 9.1 ± 6.0) and hyperplastic polyps (mean ± SD, 3.1 ± 3.7; P = 10-5 and 8 à 10-12, respectively), and for sessile serrated adenomas (mean ± SD, 20.1 ± 13.3) vs normal and hyperplastic polyps (P = .02 and 3 à 10-7, respectively). Conclusions: Claudin-1 is overexpressed in premalignant colonic lesions, and can be detected endoscopically in vivo with a near-infrared, labeled peptide. Keywords: Colon Cancer, Early Detection, Molecular Imaging

Use of a thin-film piezoelectric microactuator for axial scanning during multi-photon vertical cross-sectional imaging is described. The actuator uses thin-film lead-zirconate-titanate to generate the upward displacement of a central mirror platform micro-machined from a silicon-on-insulator wafer to dimensions compatible with endoscopic imaging instruments. Device modeling in this paper focuses on the existence of frequencies near device resonance producing vertical motion with minimal off-axis tilt, even in the presence of multiple vibration modes and non-uniformity in fabrication outcomes. Operation near rear resonance permits large stroke lengths at low voltages relative to other vertical microactuators. Highly uniform vertical motion of the mirror platform is a key requirement for vertical cross-sectional imaging in the remote scan architecture being used for multi-photon instrument prototyping. The stage is installed in a benchtop testbed in combination with an electrostatic mirror that performs in-plane scanning. Vertical sectional images are acquired from 15- $\mu \text{m}$ diameter beads and excised mouse colon tissue. [2017-0073]

We report the development, characterization, and validation of a peptide specific for the extracellular domain of HER2. This probe chemistry was developed for molecular imaging by using a structural model to select an optimal combination of amino acids that maximize the likelihood for unique hydrophobic and hydrophilic interactions with HER2 domain 3. The sequence KSPNPRF was identified and conjugated with either FITC or Cy5.5 via a GGGSK linker using Fmoc-mediated solid-phase synthesis to demonstrate flexibility for this chemical structure to be labeled with different fluorophores. A scrambled sequence was developed for control by altering the conformationally rigid spacer and moving both hydrophobic and hydrophilic amino acids on the C-terminus. We validated peptide specificity for HER2 in knockdown and competition experiments using human colorectal cancer cells in vitro, and measured a binding affinity of kd = 21 nM and time constant of k = 0.14 min(-1) (7.14 min). We used this peptide with either topical or intravenous administration in a preclinical model of colorectal cancer to demonstrate specific uptake in spontaneous adenomas and to show feasibility for real time in vivo imaging with near-infrared fluorescence. We used this peptide in immunofluorescence studies of human proximal colon specimens to evaluate specificity for sessile serrated and sporadic adenomas. Improved visualization can be used endoscopically to guide tissue biopsy and detect premalignant lesions that would otherwise be missed. Our peptide design for specificity to HER2 is promising for clinical translation in molecular imaging methods for early cancer detection.

Background and study aims: To demonstrate the clinical use of a multimodal endoscope with a targeted fluorescently labeled peptide for quantitative detection of Barrett’s neoplasia. Patients and methods: We studied 50 patients with Barrett’s esophagus using a prototype multimodal endoscope with a fluorescently labeled peptide. Co-registered fluorescence and reflectance images were converted to ratios to correct for differences in distance and geometry over the image field of view. The ratio images were segmented using a unique threshold that maximized the variance between high and low intensities to localize regions of high grade dysplasia (HGD) and esophageal adenocarcinoma (EAC). Results: Early neoplasia (HGD and EAC) was identified with 94 % specificity and 96 % positive predictive value at a threshold of 1.49. The mean results for HGD and EAC were significantly greater than those for squamous/Barrett’s esophagus and low grade dysplasia by one-way analysis of variance (ANOVA). The receiver operator characteristic curve for detection of early neoplasia had an area under the curve of 0.884. No adverse events associated with the endoscope or peptide were found. Conclusion: A multimodal endoscope can quantify fluorescence images from targeted peptides to localize early Barrett’s neoplasia. (ClinicalTrials.gov number NCT01630798.)

ErbB2 expression in early breast cancer can predict tumor aggressiveness and clinical outcomes in large patient populations. Accurate assessment with physical biopsy and conventional pathology can be limited by tumor heterogeneity. We aim to demonstrate real-time optical sectioning using a near-infrared labeled ErbB2 peptide that generates tumor-specific contrast in human xenograft breast tumors in vivo. We used IRDye800CW as the fluorophore, validated performance characteristics for specific peptide binding to cells in vitro, and investigated peak peptide uptake in tumors using photoacoustic tomography. We performed real-time optical imaging using a handheld dual-axes confocal fluorescence endomicroscope that collects light off-axis to reduce tissue scattering for greater imaging depths. Optical sections in either the vertical or horizontal plane were collected with sub-cellular resolution. Also, we found significantly greater peptide binding to pre-clinical xenograft breast cancer in vivo and to human specimens of invasive ductal carcinoma that express ErbB2 ex vivo. We used a scrambled peptide for control. Peptide biodistribution showed high tumor uptake by comparison with other organs to support safety. This novel integrated imaging strategy is promising for visualizing ErbB2 expression in breast tumors and serve as an adjunct during surgery to improve diagnostic accuracy, identify tumor margins, and stage early cancers.

We demonstrate a side-view endomicroscope using a monolithic 3-axis scanner placed in the post-objective position that performs either tilt or piston motion to achieve either optical scan angles >10° or large vertical displacements, respectively. This configuration allows for scaling down of instrument dimensions for high maneuverability and accurate positioning in vivo. Images exceeded either 700 × 600 μm2 in the horizontal plane or vertical depths of 200 μm. Resolution of 1.19 and 3.46 μm was obtained in the horizontal and oblique planes, respectively. Optical sections were collected from dysplastic colonic epithelium in vivo in mice that express tdTomato at 10 Hz to visualize individual cells.

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.

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

We present an electrostatic MEMS scanner for a dual-axes confocal fluorescence endomicroscope to collect fluorescent images in three orthogonal planes of tissue with large field of view (FOV) at high frame rate. The scanner employs a novel lever-based dual-gimbal structure to perform coupling-free switchable XY/XZ/YZ two-dimensional (2D) beam scanning, and uses the parametric resonance excitation mechanism to produce large-amplitude motions. Test results show that the scanner can achieve large optical deflection angles, up to ±14.5° in the X-axis and up to ±10° in the Y-axes, and displacements up to 370μm in the Z-axis at atmospheric pressure, and its resonant frequencies of Z-, Y-, X-axes are near at 0.54kHz, 3.47kHz and 8.55kHz, respectively.

We demonstrate vertical cross-sectional (XZ-plane) images of near-infrared (NIR) fluorescence with a handheld dual axes confocal endomicroscope that reveals specific binding of a Cy5.5-labeled peptide to pre-malignant colonic mucosa. This view is perpendicular to the tissue surface, and is similar to that used by pathologists. The scan head is 10 mm in outer diameter (OD), and integrates a one dimensional (1-D) microelectromechanical systems (MEMS) X-axis scanner and a bulky lead zirconate titanate (PZT) based Z-axis actuator. The microscope images in a raster-scanning pattern with a ±6 degrees (mechanical) scan angle at ~3 kHz in the X-axis (fast) and up to 10 Hz (0–400 μm) in the Z-axis (slow). Vertical cross-sectional fluorescence images are collected with a transverse and axial resolution of 4 and 5 μm, respectively, over a field-of-view of 800 μm (width) × 400 μm (depth). NIR vertical cross-sectional fluorescence images of fresh mouse colonic mucosa demonstrate histology-like imaging performance with this miniature instrument.

Confocal endomicroscopy is an emerging imaging technology that has recently been introduced into the clinic to instantaneously collect “optical biopsies” in vivo with histology-like quality. Here, we demonstrate a fast scanner located in the distal end of a side-viewing instrument using a compact lens assembly with numerical aperture of 0.5 to achieve a working distance of 100 μm and field-of-view of 300 × 400 μm2. The microelectromechanical systems (MEMS) mirror was designed based on the principle of parametric resonance and images at 5 frames per second. The instrument has a 4.2 mm outer diameter and 3 cm rigid length, and can pass through the biopsy channel of a medical endoscope. We achieved real time optical sections of NIR fluorescence with 0.87 μm lateral resolution, and were able to visualize in vivo binding of a Cy5.5-labeled peptide specific for EGFR to the cell surface of pre-cancerous colonocytes within the epithelium of dysplastic crypts in mouse colon. By performing targeted imaging with endomicroscopy, we can visualize molecular expression patterns in vivo that provide a biological basis for disease detection.

We present an integrated monolithic, electrostatic 3D MEMS scanner with a compact chip size of 3.2 × 2.9 mm(2). Use of parametric excitation near resonance frequencies produced large optical deflection angles up to ± 27° and ± 28.5° in the X- and Y-axes and displacements up to 510 μm in the Z-axis with low drive voltages at atmospheric pressure. When packaged in a dual axes confocal endomicroscope, horizontal and vertical cross-sectional images can be collected seamlessly in tissue with a large field-of-view of >1 × 1 mm(2) and 1 × 0.41 mm(2), respectively, at 5 frames/sec.

Hepatocellular carcinoma, the second most lethal cancer worldwide, was detected with photoacoustic imaging via an EGFR specific peptide probe in nude mice xenograft. Tumor contrast peaked at 3 hours post-injection and cleared by 24 hours.

We demonstrate a side-viewing confocal endomicroscope with an outer diameter of 4.2 mm that can image either the horizontal or vertical plane, by using an integrated monolithic, electrostatic 3D MEMS scanner.

We demonstrate a handheld multiphoton endomicroscope with 3.4 mm distal diameter that can repetitively image mouse colon in vivo. A 2D resonant MEMS mirror was developed to perform beam scanning in a Lissajous pattern. The instrument has an effective numerical aperture of 0.63, lateral and axial resolution of 2.03 and 9.02 μm, respectively, working distance of 60 μm, and image field-of-view of 300 × 300 μm(2). Hoechst was injected intravenously in mice to stain cell nuclei. We were able to collect histology-like images in vivo at 5 frames/sec, and distinguish between normal and pre-malignant colonic epithelium.

Objectives Colorectal cancer initially lies dormant as dysplasia, a premalignant state that provides an opportunity for early cancer detection. Dysplasia can be flat in morphology, focal in size, and patchy in distribution, and thus it appears "invisible" on conventional wide-field endoscopy. Aims We aim to develop and validate a peptide that is specific for epidermal growth factor receptor (EGFR), a cell surface target that is overexpressed in colonic adenomas and is readily accessible for imaging. Methods We expressed and purified the extracellular domain of EGFR for use with phage display to identify a peptide QRHKPRE that binds to domain 2 of this target. A near-infrared fluorescence endoscope was used to perform in vivo imaging to validate specific peptide binding to spontaneous colonic adenomas in a mouse model with topical administration. We also validated specific peptide binding to human colonic adenomas on immunohistochemistry and immunofluorescence. Results After labeling with Cy5.5, we validated specific peptide binding to EGFR on knockdown and competition studies. Peptide binding to cells occurred within 2.46 min and had an affinity of 50 nm. No downstream signaling was observed. We measured a target-to-background ratio of 4.0±1.7 and 2.7±0.7, for polyps and flat lesions, respectively. On immunofluorescence of human colonic specimens, greater intensity from peptide binding to dysplasia than normal was found with a 19.4-fold difference. Conclusions We have selected and validated a peptide that can be used as a specific contrast agent to identify colonic adenomas that overexpress EGFR in vivo on fluorescence endoscopy.

High frequency large scanning angle electrostatically actuated microelectromechanical systems (MEMS) mirrors are used in a variety of applications involving fast optical scanning. A 1-D parametrically resonant torsional micromirror for use in biomedical imaging is analyzed here with respect to operation by duty-cycled square waves. Duty-cycled square wave excitation can have significant advantages for practical mirror regulation and/or control. The mirror's nonlinear dynamics under such excitation is analyzed in a Hill's equation form. This form is used to predict stability regions (the voltage-frequency relationship) of parametric resonance behavior over large scanning angles using iterative approximations for nonlinear capacitance behavior of the mirror. Numerical simulations are also performed to obtain the mirror's frequency response over several voltages for various duty cycles. Frequency sweeps, stability results, and duty cycle trends from both analytical and simulation methods are compared with experimental results. Both analytical models and simulations show good agreement with experimental results over the range of duty cycled excitations tested. This paper discusses the implications of changing amplitude and phase with duty cycle for robust open-loop operation and future closed-loop operating strategies.

Many powerful drugs have limited clinical utility because of poor water solubility and high systemic toxicity. Here, we formulated a targeted nanomedicine, rapamycin encapsulated in pegylated octadecyl lithocholate micelles labeled with a new ligand for colorectal neoplasia, LTTHYKL peptide. CPC;Apc mice that spontaneously develop colonic adenomas were treated with free rapamycin, plain rapamycin micelles, and peptide-labeled rapamycin micelles via intraperitoneal injection for 35 days. Endoscopy was performed to monitor adenoma regression in vivo. We observed complete adenoma regression at the end of therapy. The mean regression rate for peptide-labeled rapamycin micelles was significantly greater than that for plain rapamycin micelles, P

Pathologists evaluate histology sectioned perpendicular to the tissue surface, or vertical cross-section. This orientation (XZ-plane) enables evaluation of mucosal differentiation in the basilar-to-luminal direction. Current endomicroscopes use a conventional (single axis) optical design.1 Imaging is limited to horizontal cross-sections (XY-plane) where the micro-anatomy is frequently similar across the field-of-view (FOV). In the dual axes configuration, light is delivered and collected off-axis, and images can be detected over a much larger range of intensities.2 Molecular images collected using fluorescence can improve specificity for disease detection and reveal functional properties about tissue.3 Proper interpretation of these images requires correlation with the micro-anatomy. We aim to demonstrate the simultaneous collection of two fluorescence images in vivo in vertical cross-sections using a dual axes confocal endomicroscope. An overlay of molecular and anatomical images from normal and dysplastic mouse colonic mucosa will be displayed in real time.