Proceedings Volume 10040

Endoscopic Microscopy XII

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Proceedings Volume 10040

Endoscopic Microscopy XII

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Volume Details

Date Published: 25 April 2017
Contents: 8 Sessions, 10 Papers, 18 Presentations
Conference: SPIE BiOS 2017
Volume Number: 10040

Table of Contents

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Table of Contents

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  • Front Matter: Volume 10040
  • New Techniques
  • Spectroscopy/Polarization
  • Tethered Capsule Endomicroscopy
  • Spectral Encoding
  • Optical Coherence Tomography
  • Novel Imaging Probes
  • MEMS-based Endoscopic Imaging: Joint Session with Conferences 10040 and 10116
Front Matter: Volume 10040
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Front Matter: Volume 10040
This PDF file contains the front matter associated with SPIE Proceedings Volume 10040, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.
New Techniques
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Differential structured illumination microendoscopy for in vivo imaging of molecular contrast agents and cervical dysplasia (Conference Presentation)
Pelham Keahey, Preetha Ramalingam, Kathleen Schmeler, et al.
Fiber-optic microendoscopes have shown promise as an imaging tool capable of visualizing molecular contrast agents used to study disease in vivo. The small size and flexibility of fiber-bundle probes make them ideal for in vivo use. However, image contrast can be severely limited when imaging highly scattering tissue. Optical sectioning techniques such as confocal or structured illumination can improve image contrast in microendoscopes by rejecting out-of-focus light generated in highly scattering tissue. However, optical sectioning techniques can reduce imaging speed or require complex opto-mechanical components to be installed on the distal end of the fiber-bundle. Here we present differential structured illumination microendoscopy (DSIMe) capable of performing structured illumination imaging in a fiber-optic microendoscope. Sectioning can be performed at video rates without the need for opto-mechanical components attached to the distal end of the fiber-bundle. Improved axial response of DSIMe is demonstrated using an optical phantom and we show image contrast enhancement in highly scattering mouse tissue imaged ex vivo. We also demonstrate contrast enhancement using DSIMe to image cervical tissue in vivo in patients diagnosed with cervical adenocarcinoma in situ and an improved ability to identify cellular changes associated with neoplasia.
Evaluation of computational endomicroscopy architectures for minimally-invasive optical biopsy
John P. Dumas, Muhammad A. Lodhi, Waheed U. Bajwa, et al.
We are investigating compressive sensing architectures for applications in endomicroscopy, where the narrow diameter probes required for tissue access can limit the achievable spatial resolution. We hypothesize that the compressive sensing framework can be used to overcome the fundamental pixel number limitation in fiber-bundle based endomicroscopy by reconstructing images with more resolvable points than fibers in the bundle. An experimental test platform was assembled to evaluate and compare two candidate architectures, based on introducing a coded amplitude mask at either a conjugate image or Fourier plane within the optical system. The benchtop platform consists of a common illumination and object path followed by separate imaging arms for each compressive architecture. The imaging arms contain a digital micromirror device (DMD) as a reprogrammable mask, with a CCD camera for image acquisition. One arm has the DMD positioned at a conjugate image plane (“IP arm”), while the other arm has the DMD positioned at a Fourier plane (“FP arm”). Lenses were selected and positioned within each arm to achieve an element-to-pixel ratio of 16 (230,400 mask elements mapped onto 14,400 camera pixels). We discuss our mathematical model for each system arm and outline the importance of accounting for system non-idealities. Reconstruction of a 1951 USAF resolution target using optimization-based compressive sensing algorithms produced images with higher spatial resolution than bicubic interpolation for both system arms when system non-idealities are included in the model. Furthermore, images generated with image plane coding appear to exhibit higher spatial resolution, but more noise, than images acquired through Fourier plane coding.
Chip-on-the-tip ultra-compact flexible endoscopic epifluorescence video-microscope for in-vivo imaging in medical and biomedical fields
We demonstrate a flexible stand-alone, minimally invasive video-endomicroscope with an outer diameter of 1.6 mm and a length of the rigid tip of 6.7 mm that enables surgeons and biologists to image hardly accessible regions in-vivo in epifluorescence mode. The 60 mg light device improves state-of-the-art objectives by a double deflection approach using a side-fire fiber in combination with spherical microlenses, GRIN-lenses with a specific adapted gradient index profile and an extremely miniaturized chip-on-the-tip camera to achieve an excellent imaging quality. A high NA of 0.7 enables the observation of subcellular features within the entire field of view with a diameter of 183 μm, assure a bright and high-contrast image and promise a good overview during the intervention. Ex-vivo measurements of biological samples confirmed the functionality of the probe.
Spectroscopy/Polarization
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Optical characterization and polarization calibration for rigid endoscopes
Missael Garcia, Viktor Gruev
Polarization measurements give orthogonal information to spectral images making them a great tool in the characterization of environmental parameters in nature. Thus, polarization imagery has proven to be remarkably useful in a vast range of biomedical applications. One such application is the early diagnosis of flat cancerous lesions in murine colorectal tumor models, where polarization data complements NIR fluorescence analysis. Advances in nanotechnology have led to compact and precise bio-inspired imaging sensors capable of accurately co-registering multidimensional spectral and polarization information. As more applications emerge for these imagers, the optics used in these instruments get very complex and can potentially compromise the original polarization state of the incident light. Here we present a complete optical and polarization characterization of three rigid endoscopes of size 1.9mm x 10cm (Karl Storz, Germany), 5mm x 30cm, and 10mm x 33cm (Olympus, Germany), used in colonoscopy for the prevention of colitis-associated cancer. Characterization results show that the telescope optics act as retarders and effectively depolarize the linear component. These incorrect readings can cause false-positives or false-negatives leading to an improper diagnosis. In this paper, we offer a polarization calibration scheme for these endoscopes based on Mueller calculus. By modeling the optical properties from training data as real-valued Mueller matrices, we are able to successfully reconstruct the initial polarization state acquired by the imaging system.
Minimally invasive optical biopsy for oximetry
Marieke A. van der Putten, James M. Brewer, Andrew R. Harvey
The study of localised oxygen saturation in blood vessels can shed light on the etiology and progression of many diseases with which hypoxia is associated. For example, hypoxia in the tendon has been linked to early stages of rheumatoid arthritis, an auto-immune inflammatory disease. Vascular oximetry of deep tissue presents significant challenges as vessels are not optically accessible. In this paper, we present a novel multispectral imaging technique for vascular oximetry, and recent developments made towards its adaptation for minimally invasive imaging. We present proof-of-concept of the system and illumination scheme as well as the analysis technique. We present results of a validation study performed in vivo on mice with acutely inflamed tendons. Adaptation of the technique for minimally invasive microendoscopy is also presented, along with preliminary results of minimally invasive ex vivo vascular oximetry.
Spectral and lifetime endomicroscopic measurements using one and two-photon excitation
A. Ibrahim, F. Poulon, M. Zanello, et al.
Current surgical biopsy needs several days for the analysis process to be finished. Anatomopathologists provide analysis reports to the surgeon a few days after the surgical intervention, which makes it a lengthy decision making practice. In addition, the lack of precise guidance often leads to inaccuracies in the selection of tissue regions for biopsy and so necessitates repeating the operation sometimes. Our project aims at reducing this time as well as patient discomfort. In this context, we propose to develop a multimodal nonlinear endomicroscope providing several means of contrast. Among these contrast that are useful in the detection of tumor regions, we note imaging by linear and non-linear fluorescence, by second and third harmonic generation and by reflectance. In addition, this technique allows fluorescence lifetime and spectral measurements. Our endomicroscopic system is based on a new homemade customized double-clad photonic crystal fiber (DC-PCF). Finally, this double-clad micro structured optical fiber insures visible and near infrared excitation. This system was tested by measuring fluorescence lifetime and the spectral shape of a fixed tumoral brain sample in one and two photon excitations.
Tethered Capsule Endomicroscopy
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Feasibility study of Tethered Capsule Endomicroscopy (TCE) deployment in the small intestine (Conference Presentation)
Environmental enteric dysfunction (EED) is a poorly understood disease of the small intestine that causes nutrient malabsorption in children, predominantly from low and middle income countries. The clinical importance of EED is neurological and growth stunting that remains as the child grows into adulthood. Tethered capsule endomicroscopy (TCE) has the potential to improve the understanding of EED and could be used to determine the effectiveness of EED interventions. TCE in the adult esophagus and the duodenum has been demonstrated for Barrett`s esophagus and celiac disease diagnosis, respectively. While adult subjects can independently swallow these capsules, it is likely that infants will not, and, as a result, new strategies for introducing these devices in young children aged 0.5-2 years need to be investigated. Our first approach will be to introduce the TCE devices in infants under the aid of endoscopic guidance. To determine the most effective method, we have tested endoscopic approaches for introducing TCE devices into the small intestine of living swine. These methods will be compared and contrasted to discuss the most effective means for endoscopic tethered capsule introduction into the small intestine.
Optimizing the villi visualization by tethered capsule OCT endomicroscopy for comprehensive imaging of human duodenum (Conference Presentation)
Celiac disease (CD) affects around 1% of the global population and can cause serious long-term symptoms including malnutrition, fatigue, and diarrhea, amongst others. Despite this, it is often left undiagnosed. Currently, a tissue diagnosis of CD is made by random endoscopic biopsy of the duodenum to confirm the existence of microscopic morphologic alterations in the intestinal mucosa. However, duodenal endoscopic biopsy is problematic because the morphological changes can be focal and endoscopic biopsy is plagued by sampling error. Additionally, tissue artifacts can also an issue because cuts in the transverse plane can make duodenal villi appear artifactually shortened and can bias the assessment of intraepithelial inflammation. Moreover, endoscopic biopsy is costly and poorly tolerated as the patient needs to be sedated to perform the procedure. Our lab has previously developed technology termed tethered capsule OCT endomicroscopy (TCE) to overcome these diagnostic limitations of endoscopy. TCE involves swallowing an optomechanically-engineered pill that generates 3D images of the GI tract as it traverses the lumen of the organ via peristalsis, assisted by gravity. In several patients we have demonstrated TCE imaging of duodenal villi, however the current TCE device design is not optimal for CD diagnosis as the villi compress when in contact with the smooth capsule’s wall. In this work, we present methods for structuring the outer surface of the capsule to improve the visualization of the villi height and crypt depth. Preliminary results in humans suggest that new TCE capsule enables better visualization of villous architecture, making it possibly to comprehensively scan the entire duodenum to obtain a more accurate tissue diagnosis of CD.
Tethered SECM endoscopic capsule for the diagnosis of eosinophilic esophagitis (Conference Presentation)
Dukho Do, DongKyun Kang, Nima Tabatabaei, et al.
Eosinophilic Esophagitis (EoE) is an inflammatory disease caused by inhaled or ingested food allergies, and characterized by the infiltration of eosinophils in the esophagus. The gold standard for diagnosing EoE is to conduct endoscopy and obtain multiple biopsy specimens from different portions of the esophagus; an exam is considered positive if more than 15 eosinophils per high power field (HPF) in any of the biopsies. This method of diagnosis is problematic because endoscopic biopsy is expensive and poorly tolerated and the esophageal eosinophil burden needs to be monitored frequently during the course of the disease. Spectrally encoded confocal microscopy (SECM) is a high-speed confocal microscopy technology that can visualize individual eosinophils in large microscopic images of the human esophagus, equivalent to more than 30,000 HPF. Previously, we have demonstrated that tethered capsule SECM can be conducted in unsedated subjects with diagnosed EoE. However, speckle noise and the relatively low resolution in images obtained with the first capsule prototypes made it challenging to distinguish eosinophils from other cells. In this work, we present a next-generation tethered SECM capsule, which has been modified to significantly improve image quality. First, we substituted the single mode fiber with a dual-clad fiber to reduce speckle noise. A gradient-index multimode fiber was fusion spliced at the tip of the dual-clad fiber to increase the effective numerical aperture of the fiber from 0.09 to 0.15, expanding the beam more rapidly to increase the illumination aperture at the objective. These modifications enabled the new SECM capsule to achieve a lateral resolution of 1.8 µm and an axial resolution of 16.1 µm, which substantially improves the capacity of this probe to visualize cellular features in human tissue. The total size of the SECM capsule remained 6.75 mm in diameter and 31 mm in length. We are now in the process of testing this new SECM capsule in humans. Early results using this new SECM capsule suggest that this technology has the potential to be an effective tool for the diagnosis of EoE.
Single lens system for forward-viewing navigation and scanning side-viewing optical coherence tomography
The optical design for a dual modality endoscope based on piezo scanning fiber technology is presented including a novel technique to combine forward-viewing navigation and side viewing OCT. Potential applications include navigating body lumens such as the fallopian tube, biliary ducts and cardiovascular system. A custom cover plate provides a rotationally symmetric double reflection of the OCT beam to deviate and focus the OCT beam out the side of the endoscope for cross-sectional imaging of the tubal lumen. Considerations in the choice of the scanning fiber are explored and a new technique to increase the divergence angle of the scanning fiber to improve system performance is presented. Resolution and the necessary scanning density requirements to achieve Nyquist sampling of the full image are considered. The novel optical design lays the groundwork for a new approach integrating side-viewing OCT into multimodality endoscopes for small lumen imaging. KEYWORDS:
Spectral Encoding
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In vivo microscopy of human leucocytes (Conference Presentation)
Matan Winer, Daniella Yeheskely-Hayon, Adel Zeidan, et al.
White blood cells (WBC) analysis is an important part of the complete blood count, providing good indication of the patient’s immune system status. The most common types of WBCs are the neutrophils and lymphocytes that comprise approximately 60% and 30% of the total WBC count, respectively; differentiating between these cells at the point of care would assist in accurate diagnosis of the possible source of infection (viral or bacterial) and in effective prescription of antibiotics. In this work, we demonstrate the potential of spectrally encoded flow cytometry (SEFC) to non-invasively image WBC in human patients, allowing morphology characterization of the main types of WBCs. The optical setup includes a broadband light that was diffracted and focused onto a single transverse line within the cross section of a small blood vessel at the inner patient lip. Light backscattered from the tissue was measured by a high-speed spectrometer, forming a two-dimensional reflectance confocal image of the flowing cells. By imaging at different depths into vessels of different diameters, we determine optimal imaging conditions (i.e. imaging geometry, speed and depth) for counting the total amount of WBCs and for differentiating between their main types. The presented technology could serve for analyzing the immune system status at the point of care, and for studying the morphological and dynamical characteristics of these cells in vivo.
Measuring sickle cell morphology in flow using spectrally encoded flow cytometry (Conference Presentation)
Inna Kviatkovsky, Adel Zeidan, Daniella Yeheskely-Hayon, et al.
During a sickle cell crisis in sickle cell anemia patients, deoxygenated red blood cells may change their mechanical properties and block small blood vessels, causing pain, local tissue damage and even organ failure. Measuring these cellular structural and morphological changes is important for understanding the factors contributing to vessel blockage and developing an effective treatment. In this work, we use spectrally encoded flow cytometry for confocal, high-resolution imaging of flowing blood cells from sickle cell anemia patients. A wide variety of cell morphologies were observed by analyzing the interference patterns resulting from reflections from the front and back faces of the cells’ membrane. Using numerical simulation for calculating the two-dimensional reflection pattern from the cells, we propose an analytical expression for the three-dimensional shape of a characteristic sickle cell and compare it to a previous from the literature. In vitro spectrally encoded flow cytometry offers new means for analyzing the morphology of sickle cells in stress-free environment, and could provide an effective tool for studying the unique physiological properties of these cells.
Ultraminiature video-rate forward-view spectrally encoded endoscopy with straight axis configuration
Zhuo Wang, Tzu-Yu Wu, Mark A. Hamm, et al.
As one of the smallest endoscopes that have been demonstrated, the spectrally encoded endoscope (SEE) shows potential for the use in minimally invasive surgeries. While the original SEE is designed for side-view applications, the forwardview (FV) scope is more desired by physicians for many clinical applications because it provides a more natural navigation. Several FV SEEs have been designed in the past, which involve either multiple optical elements or one optical element with multiple optically active surfaces. Here we report a complete FV SEE which comprises a rotating illumination probe within a drive cable, a sheath and a window to cover the optics, a customized spectrometer, hardware controllers for both motor control and synchronization, and a software suite to capture, process and store images and videos. In this solution, the optical axis is straight and the dispersion element, i.e. the grating, is designed such that the slightly focused light after the focusing element will be dispersed by the grating, covering forward view angles with high diffraction efficiencies. As such, the illumination probe is fabricated with a diameter of only 275 μm. The twodimensional video-rate image acquisition is realized by rotating the illumination optics at 30 Hz. In one finished design, the scope diameter including the window assembly is 1.2 mm.
Optical Coherence Tomography
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Extended depth of focus tethered capsule OCT endomicroscopy for upper gastrointestinal tract imaging (Conference Presentation)
Endoscopy, the current standard of care for the diagnosis of upper gastrointestinal (GI) diseases, is not ideal as a screening tool because it is costly, necessitates a team of medically trained personnel, and typically requires that the patient be sedated. Endoscopy is also a superficial macroscopic imaging modality and therefore is unable to provide detailed information on subsurface microscopic structure that is required to render a precise tissue diagnosis. We have overcome these limitations through the development of an optical coherence tomography tethered capsule endomicroscopy (OCT-TCE) imaging device. The OCT-TCE device has a pill-like form factor with an optically clear wall to allow the contained opto-mechanical components to scan the OCT beam along the circumference of the esophagus. Once swallowed, the OCT-TCE device traverses the esophagus naturally via peristalsis and multiple cross-sectional OCT images are obtained at 30-40 μm lateral resolution by 7 μm axial resolution. While this spatial resolution enables differentiation of squamous vs columnar mucosa, crucial microstructural features such as goblet cells (~10 μm), which signify intestinal metaplasia in BE, and enlarged nuclei that are indicative of dysplasia cannot be resolved with the current OCT-TCE technology. In this work we demonstrate a novel design of a high lateral resolution OCT-TCE device with an extended depth of focus (EDOF). The EDOF is created by use of self-imaging wavefront division multiplexing that produces multiple focused modes at different depths into the sample. The overall size of the EDOF TCE is similar to that of the previous OCT-TCE device (~ 11 mm by 26 mm) but with a lateral resolution of ~ 8 μm over a depth range of ~ 2 mm. Preliminary esophageal and intestinal imaging using these EDOF optics demonstrates an improvement in the ability to resolve tissue morphology including individual glands and cells. These results suggest that the use of EDOF optics may be a promising avenue for increasing the accuracy of OCT-TCE for the diagnosis of upper GI diseases.
Miniature multimodal endoscopic probe based on double-clad fiber
Xianjin Dai, Hao Yang, Jianbo Tang, et al.
Optical coherence tomography (OCT) can obtain light scattering properties with a high resolution, while photoacoustic imaging (PAI) is ideal for mapping optical absorbers in biological tissues, and ultrasound (US) could penetrate deeply into tissues and provide elastically structural information. It is attractive and challenging to integrate these three imaging modalities into a miniature probe, through which, both optical absorption and scattering information of tissues as well as deep-tissue structure can be obtained. Here, we present a novel side-view probe integrating PAI, OCT and US imaging based on double-clad fiber which is used as a common optical path for PAI (light delivery) and OCT (light delivery/detection), and a 40 MHz unfocused ultrasound transducer for PAI (photoacoustic detection) and US (ultrasound transmission/receiving) with an overall diameter of 1.0 mm. Experiments were conducted to demonstrate the capabilities of the integrated multimodal imaging probe, which is suitable for endoscopic imaging and intravascular imaging.
Novel Imaging Probes
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High resolution microphotonic needle for endoscopic imaging (Conference Presentation)
Mohammad Amin Tadayon, Aseema Mohanty, Samantha P Roberts, et al.
GRIN (Graded index) lens have revolutionized micro endoscopy enabling deep tissue imaging with high resolution. The challenges of traditional GRIN lenses are their large size (when compared with the field of view) and their limited resolution. This is because of the relatively weak NA in standard graded index lenses. Here we introduce a novel micro-needle platform for endoscopy with much higher resolution than traditional GRIN lenses and a FOV that corresponds to the whole cross section of the needle. The platform is based on polymeric (SU-8) waveguide integrated with a microlens micro fabricated on a silicon substrate using a unique molding process. Due to the high index of refraction of the material the NA of the needle is much higher than traditional GRIN lenses. We tested the probe in a fluorescent dye solution (19.6 µM Alexa Flour 647 solution) and measured a numerical aperture of 0.25, focal length of about 175 µm and minimal spot size of about 1.6 µm. We show that the platform can image a sample with the field of view corresponding to the cross sectional area of the waveguide (80x100 µm2). The waveguide size can in principle be modified to vary size of the imaging field of view. This demonstration, combined with our previous work demonstrating our ability to implant the high NA needle in a live animal, shows that the proposed system can be used for deep tissue imaging with very high resolution and high field of view.
Focus scanning with feedback control for fiber-optic nonlinear endomicroscopy (Conference Presentation) (Withdrawal Notice)
Publisher’s Note: This conference presentation, originally published on 19 April 2017, was withdrawn per author request.
High-resolution image reconstruction for GRIN rod lens probe (Conference Presentation)
Graded-index (GRIN) lenses have been widely used for developing compact imaging devices due to the small dimensions and simple optics designs. GRIN lenses, however, have intrinsic aberration which causes a distortion of the image and thus are subject to limited resolution and blurred imaging quality. Here, we employ the high-precision wavefront measurement technique for compensation of the distortion of a GRIN lens to obtain a high-resolution and high-contrast image. In doing so, we demonstrate a high-resolution and ultra-thin endo-microscope using a GRIN. A reflection-type interferometric microscope through a GRIN lens was constructed using multiple lasers (473 nm, 532 nm, and 633 nm) as light sources. The characteristics of the aberration of the GRIN lens were measured using the digital holographic method. The distortion of the GRIN lens was removed by numerical image processing with the prior information from the pre-calibration. We apply this technique to a reflection image of biological tissues acquired by our custom-built GRIN lens probe. Consequently, a diffraction limited lateral resolution as well as improved axial resolution can be achieved. Our approach will facilitate the use of GRIN lenses for compact imaging devices without compromising optical resolution and image quality.
Common path endoscopic probes for optical coherence tomography (Conference Presentation)
Kanwarpal Singh, Joseph A. Gardecki, Guillermo J. Tearney
Background: Dispersion imbalance and polarization mismatch between the reference and sample arm signals can lead to image quality degradation in optical coherence tomography (OCT). One approach to reduce these image artifacts is to employ a common-path geometry in fiber-based probes. In this work, we report an 800 um diameter all-fiber common-path monolithic probe for coronary artery imaging where the reference signal is generated using an inline fiber partial reflector. Methods: Our common-path probe was designed for swept-source based Fourier domain OCT at 1310 nm wavelength. A face of a coreless fiber was coated with gold and spliced to a standard SMF-28 single mode fiber creating an inline partial reflector, which acted as a reference surface. The other face of the coreless fiber was shaped into a ball lens for focusing. The optical elements were assembled within a 560 µm diameter drive shaft, which was attached to a rotary junction. The drive shaft was placed inside a transparent sheath having an outer diameter of 800 µm. Results: With a source input power of 30mW, the inline common-path probe achieved a sensitivity of 104 dB. Images of human finger skin showed the characteristic layers of skin as well as features such as sweat ducts. Images of coronary arteries ex vivo obtained with this probe enabled visualization of the characteristic architectural morphology of the normal artery wall and known features of atherosclerotic plaque. Conclusion: In this work, we have demonstrated a common path OCT probe for cardiovascular imaging. The probe is easy to fabricate, will reduce system complexity and overall cost. We believe that this design will be helpful in endoscopic applications that require high resolution and a compact form factor.
MEMS-based Endoscopic Imaging: Joint Session with Conferences 10040 and 10116
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A handheld MEMS-based line-scanned dual-axis confocal microscope for early cancer detection and surgical guidance (Conference Presentation)
Ye Chen, Chengbo Yin, Linpeng Wei, et al.
Considerable efforts have been recently undertaken to develop miniature optical-sectioning microscopes for in vivo microendoscopy and point-of-care pathology. These devices enable in vivo interrogation of disease as a real-time and noninvasive alternative to gold-standard histopathology, and therefore could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Regardless of the specific modality, various trade-offs in size, speed, field of view, resolution, contrast, and sensitivity are necessary to optimize a device for a particular application. Here, a miniature MEMS-based line-scanned dual-axis confocal (LS-DAC) microscope, with a 12-mm diameter distal tip, has been developed for point-of-care pathology. The dual-axis architecture has demonstrated superior rejection of out-of-focus and multiply scattered photons compared to a conventional single-axis confocal configuration. The use of line scanning enables fast frame rates (≥15 frames/sec), which mitigates motion artifacts of a handheld device during clinical use. We have developed a method to actively align the illumination and collection beams in this miniature LS-DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo application, enables the device to achieve an axial and lateral resolution of 2.0 and 1.1 microns, respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate that this high-speed LS-DAC microscope can achieve high-contrast imaging of fluorescently labeled tissues with sufficient sensitivity for applications such as oral cancer detection and guiding brain-tumor resections.
Targeted sections in either XY or XZ plane with dual-axes confocal endomicroscope (Conference Presentation)
Gaoming Li, Haijun Li, Xiyu Duan, et al.
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.
MEMS-based side-view endomicroscope for in vivo small animal imaging (Conference Presentation)
Xiyu Duan, Haijun Li, Gaoming Li, et al.
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.
A MEMS scanner with lateral and axial scanning capability for dual axes confocal endomicroscopic in-vivo imaging (Conference Presentation)
Haijun Li, Gaoming Li, Xiyu Duan, et al.
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.