[Show abstract][Hide abstract] ABSTRACT: The inadequacy of white-light cystoscopy to detect flat bladder tumors is well recognized. Great interest exists in developing other imaging technologies to augment or supplant conventional cystoscopy. Fibered confocal microscopy offers the promise of providing in vivo histopathologic information to help distinguish malignant from benign bladder lesions. We report the initial use of this technology to visualize tumors in the human bladder.
We performed ex vivo fibered confocal imaging of fresh radical cystectomy specimens using the Mauna Kea Technologies Cellvizio system. The findings were compared with results from standard histopathology.
The bladders of four patients were imaged using the fibered confocal microscope. Normal and neoplastic urothelium manifested differences in cellular and vascular density.
This study demonstrates the feasibility of using fibered confocal microscopy to detect histologic differences between normal and neoplastic urothelium, and establishes a foundation for the use of fiber-based confocal microscopy in clinical studies.
Journal of endourology / Endourological Society 02/2009; 23(2):197-201. · 1.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We present a handheld dual-axes confocal microscope that is based on a two-dimensional microelectromechanical systems (MEMS) scanner. It performs reflectance and fluorescence imaging at 488 nm wavelength, with three-dimensional imaging capability. The fully packaged microscope has a diameter of 10 mm and acquires images at 4 Hz frame rate with a maximum field of view of 400 microm x 260 microm. The transverse and axial resolutions of the handheld probe are 1.7 microm and 5.8 microm, respectively. Capability to perform real time small animal imaging is demonstrated in vivo in transgenic mice.
[Show abstract][Hide abstract] ABSTRACT: A combination of targeted probes and new imaging technologies provides a powerful set of tools with the potential to improve the early detection of cancer. To develop a probe for detecting colon cancer, we screened phage display peptide libraries against fresh human colonic adenomas for high-affinity ligands with preferential binding to premalignant tissue. We identified a specific heptapeptide sequence, VRPMPLQ, which we synthesized, conjugated with fluorescein and tested in patients undergoing colonoscopy. We imaged topically administered peptide using a fluorescence confocal microendoscope delivered through the instrument channel of a standard colonoscope. In vivo images were acquired at 12 frames per second with 50-microm working distance and 2.5-microm (transverse) and 20-microm (axial) resolution. The fluorescein-conjugated peptide bound more strongly to dysplastic colonocytes than to adjacent normal cells with 81% sensitivity and 82% specificity. This methodology represents a promising diagnostic imaging approach for the early detection of colorectal cancer and potentially of other epithelial malignancies.
Nature medicine 05/2008; 14(4):454-8. · 28.05 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A combination of targeted probes and new imaging technologies provides a powerful set of tools with the potential to improve the early detection of cancer. To develop a probe for detecting colon cancer, we screened phage display peptide libraries against fresh human colonic adenomas for high-affinity ligands with preferential binding to premalignant tissue. We identified a specific heptapeptide sequence, VRPMPLQ, which we synthesized, conjugated with fluorescein and tested in patients undergoing colonoscopy. We imaged topically administered peptide using a fluorescence confocal microendoscope delivered through the instrument channel of a standard colonoscope. In vivo images were acquired at 12 frames per second with 50-lm working distance and 2.5-lm (transverse) and 20-lm (axial) resolution. The fluorescein-conjugated peptide bound more strongly to dysplastic colonocytes than to adjacent normal cells with 81% sensitivity and 82% specificity. This
Nature Medicine 04/2008; 14(5):585-585. · 28.05 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Optical coherence tomography (OCT) is an emerging medical imaging technology which generates high resolution, cross-sectional images in situ, without the need for excisional biopsy. Previous clinical studies using endoscopic OCT with standard 10-15 microm axial resolution have demonstrated its capability in diagnosing Barrett's esophagus (BE) and high-grade dysplasia (HGD). Quantitative OCT image analysis has shown promise for detecting HGD in Barrett's esophagus patients. We recently developed an endoscopic OCT system with an improved axial resolution of approximately 5 microm. The goal in this manuscript is to compare standard resolution OCT and ultrahigh resolution OCT (UHR-OCT) for image quality and computer-aided detection using normal and Barrett's esophagus. OCT images of gastrointestinal (GI) tissues were obtained using UHR-OCT (5.5 microm) and standard resolution OCT (13 microm). Image quality including the speckle size and sharpness was compared. Texture features of endoscopic OCT images from normal and Barrett's esophagus were extracted using quantitative metrics including spatial frequency analysis and statistical texture analysis. These features were analyzed using principal component analysis (PCA) to reduce the vector dimension and increase the discriminative power, followed by linear discrimination analysis (LDA). UHR-OCT images of GI tissues improved visualization of fine architectural features compared to standard resolution OCT. In addition, the quantitative image feature analysis showed enhanced discrimination of normal and Barrett's esophagus with UHR-OCT. The ability of UHR-OCT to resolve tissue morphology at improved resolution enables visualization of subtle features in OCT images, which may be useful in disease diagnosis. Enhanced classification of image features using UHR-OCT promises to help in the computer-aided diagnosis of GI diseases.
[Show abstract][Hide abstract] ABSTRACT: Colorectal carcinoma continues to be a leading cause of cancer morbidity and mortality despite widespread adoption of screening methods. Targeted detection and therapy using recent advances in our knowledge of in vivo cancer biomarkers promise to significantly improve methods for early detection, risk stratification, and therapeutic intervention. The behavior of molecular targets in transformed tissues is being comprehensively assessed using new techniques of gene expression profiling and high throughput analyses. The identification of promising targets is stimulating the development of novel molecular probes, including significant progress in the field of activatable and peptide probes. These probes are being evaluated in small animal models of colorectal neoplasia and recently in the clinic. Furthermore, innovations in optical imaging instrumentation are resulting in the scaling down of size for endoscope compatibility. Advances in target identification, probe development, and novel instruments are progressing rapidly, and the integration of these technologies has a promising future in molecular medicine.
Cancer biomarkers: section A of Disease markers 02/2008; 4(6):329-40. · 1.19 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Histologic interpretation of disease currently is performed with static images of excised tissues, and is limited by processing artifact, sampling error, and interpretive variability. The aim of this study was to show the use of functional optical imaging of viable mucosa for quantitative evaluation of colonic neoplasia in real time.
Fluorescein (5 mg/mL) was administered topically in 54 human subjects undergoing screening colonoscopy. Fluorescence images were collected with 488-nm excitation at 12 frames/s with the confocal microendoscopy system. Movement of fluorescein in the transient period (<5 s) and the lamina propria:crypt contrast ratio in the steady-state phase (>5 s) were quantified.
Normal mucosa showed circular crypts with uniform size, hyperplasia revealed proliferative glands with serrated lumens, and adenomas displayed distorted elongated glands. For t less than 5 seconds, fluorescein passed through normal epithelium with a peak speed of 1.14 +/- 0.09 microm/s at t = 0.5 seconds, and accumulated into lamina propria as points of fluorescence that moved through the interglandular space with an average speed of 41.7 +/- 3.4 microm/s. Passage of fluorescein through adenomatous mucosa was delayed substantially. For t greater than 5 seconds, high sensitivity, specificity, and accuracy was achieved using a discriminant function to evaluate the contrast ratio to distinguish normal from lesional mucosa (91%, 87%, and 89%, respectively; P < .001), hyperplasia from adenoma (97%, 96%, and 96%, respectively; P < .001), and tubular from villous adenoma (100%, 92%, and 93%, respectively; P < .001).
Confocal imaging can be performed in vivo to assess the functional behavior of tissue in real time for providing pathologic interpretation, representing a new method for histologic evaluation.
Clinical gastroenterology and hepatology: the official clinical practice journal of the American Gastroenterological Association 11/2007; 5(11):1300-5. · 5.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Institutional review board approval at the participating institutions was obtained. Informed consent was waived for this HIPAA-compliant study. The study purpose was to establish the correspondence of optical coherence tomographic (OCT) image findings with histopathologic findings to understand which features characteristic of breast lesions can be visualized with OCT. Imaging was performed in 119 specimens from 35 women aged 29-81 years with 3.5-microm axial resolution and 6-microm transverse resolution at 1.1-microm wavelength on freshly excised specimens of human breast tissue. Three-dimensional imaging was performed in 43 specimens from 23 patients. Microstructure of normal breast parenchyma, including glands, lobules, and lactiferous ducts, and stromal changes associated with infiltrating cancer were visible. Fibrocystic changes and benign fibroadenomas were identified. Imaging of ductal carcinoma in situ, infiltrating cancer, and microcalcifications correlated with corresponding histopathologic findings. OCT is potentially useful for visualization of breast lesions at a resolution greater than that of currently available clinical imaging methods.
[Show abstract][Hide abstract] ABSTRACT: Here we describe a simple optical design for a MEMS-based dual-axes fiber optic confocal scanning microscope that has been miniaturized for handheld imaging of tissues, and which is capable of being further scaled to smaller dimensions for endoscope compatibility while preserving it's field-of-view (FOV), working distance, and resolution. Based on the principle of parallel beams that are focused by a single parabolic mirror to a common point, the design allows the use of replicated optical components mounted and aligned within a rugged cylindrical housing that is designed for use as a handheld tissue microscope. A MEMS scanner is used for high speed scanning in the X-Y plane below the tissue surface. An additional design feature is a mechanism for controlling a variable working distance, thus producing a scan in the Z direction and allowing capture of 3-D volumetric images of tissue. The design parameters that affect the resolution, FOV, and working distance are analyzed using ASAP TM optical modeling software and verified by experimental results. Other features of this design include use of a solid immersion lens (SIL), which enhances both resolution and FOV, and also provides index matching between the optics and the tissue.
[Show abstract][Hide abstract] ABSTRACT: We demonstrate a dual-axes confocal reflectance microscope in a 10-mm diameter handheld package. Miniaturization is achieved by using a barbell-shaped, gimbaled, two-dimensional MEMS scanner that is actuated by self-aligned vertical comb actuators. The maximum DC optical scan angles are plusmn3.3deg on the inner axis and plusmn1.0deg on the outer axis, and the corresponding resonance frequencies are 3.54 kHz and 1.1 kHz. The imaging rate is 30 frames/second, and the microscope achieves full-width-half-maximum (FWHM) transverse and axial resolutions of 8 mum and 10 mum, respectively
Optical MEMS and Their Applications Conference, 2006. IEEE/LEOS International Conference on; 09/2006
[Show abstract][Hide abstract] ABSTRACT: Ultrahigh-resolution optical coherence tomography (OCT) has an axial resolution of <5 microm, 2 to 3 times finer than standard OCT. This study investigates ultrahigh-resolution and three-dimensional OCT for ex vivo imaging of the large and small intestines and correlates images with histology.
Ultrahigh-resolution OCT imaging was performed on fresh surgical specimens from the large and small intestines in the pathology laboratory, and images were correlated with histology. OCT was performed at 1.3-microm wavelength with 4.5-microm axial x 11-microm transverse resolution and at 1.1-microm wavelength with 3.5-microm axial x 6-microm transverse resolution. Three-dimensional OCT also was investigated.
Normal and pathologic areas from 23 surgical specimens of the large and small intestines were imaged. Ultrahigh-resolution OCT distinguished the epithelial layer of the mucosa and visualized individual villi, glands, and crypts. Finer transverse resolutions improved visualization of features, e.g., the epithelium, but reduced the depth of field. Architectural distortion of glands from inflammatory and neoplastic processes was observed. Three-dimensional rendering enabled visualization of surface pit pattern and mucosal folds as well as subsurface crypt microstructure.
This study evaluates new OCT technology and can provide a baseline for interpreting future ultrahigh-resolution endoscopic OCT studies.
[Show abstract][Hide abstract] ABSTRACT: High performance, short coherence length light sources with broad bandwidths and high output powers are critical for high speed, ultrahigh resolution OCT imaging. We demonstrate an all-fiber continuous-wave Raman light source based on a photonic crystal fiber, pumped by a continuous-wave Yb-fiber laser, which generates 330 mW output power and 140 nm bandwidths. The light source is compact, robust, turnkey and requires no optical alignment. In vivo high speed, ultrahigh resolution OCT imaging of tissues with < 5 µm axial resolution at 1.3 µm center wavelength is demonstrated.
[Show abstract][Hide abstract] ABSTRACT: Real-time, ultrahigh-resolution optical coherence tomography (OCT) is demonstrated in the 1.4-1.7-microm wavelength region with a stretched-pulse, passively mode-locked, Er-doped fiber laser and highly nonlinear fiber. The fiber laser generates 100-mW, linearly chirped pulses at a 51-MHz repetition rate. The pulses are compressed and then coupled into a normally dispersive highly nonlinear fiber to generate a low-noise supercontinuum with a 180-nm FWHM bandwidth and 38 mW of output power. This light source is stable, compact, and broadband, permitting high-speed, real-time, high-resolution OCT imaging. In vivo high-speed OCT imaging of human skin with approximately 5.5-microm resolution and 99-dB sensitivity is demonstrated.
[Show abstract][Hide abstract] ABSTRACT: Early detection of gastrointestinal cancer is essential for the patient
treatment and medical care. Endoscopically guided biopsy is currently
the gold standard for the diagnosis of early esophageal cancer, but can
suffer from high false negative rates due to sampling errors. Optical
coherence tomography (OCT) is an emerging medical imaging technology
which can generate high resolution, cross-sectional images of tissue in
situ and in real time, without the removal of tissue specimen. Although
endoscopic OCT has been used successfully to identify certain
pathologies in the gastrointestinal tract, the resolution of current
endoscopic OCT systems has been limited to 10 - 15 m for clinical
procedures. In this study, in vivo imaging of the gastrointestinal tract
is demonstrated at a three-fold higher resolution (< 5 m), using a
portable, broadband, Cr4+:Forsterite laser as the optical
light source. Images acquired from the esophagus, gastro-esophageal
junction and colon on animal model display tissue microstructures and
architectural details at high resolution, and the features observed in
the OCT images are well-matched with histology. The clinical feasibility
study is conducted through delivering OCT imaging catheter using
standard endoscope. OCT images of normal esophagus, Barrett's esophagus,
and esophageal cancers are demonstrated with distinct features. The
ability of high resolution endoscopic OCT to image tissue morphology at
an unprecedented resolution in vivo would facilitate the development of
OCT as a potential imaging modality for early detection of neoplastic
[Show abstract][Hide abstract] ABSTRACT: Ultrahigh resolution optical coherence tomography (OCT) is an emerging imaging modality that enables noninvasive imaging of tissue with 1- to 3-microm resolutions. Initial OCT studies have typically been performed using harvested tissue specimens (ex vivo). No reports have investigated postexcision tissue degradation on OCT image quality. We investigate the effects of formalin fixation and commonly used cell culture media on tissue optical scattering characteristics in OCT images at different times postexcision compared to in vivo conditions. OCT imaging at 800-nm wavelength with 1.5-mum axial resolution is used to image the hamster cheek pouch in vivo, followed by excision and imaging during preservation in phosphate-buffered saline (PBS), Dulbecco's Modified Eagle's Media (DMEM), and 10% neutral-buffered formalin. Imaging is performed in vivo and at sequential time points postexcision from 15 min to 10 to 18 h. Formalin fixation results in increases in scattering intensity from the muscle layers, as well as shrinkage of the epithelium, muscle, and connective tissue of approximately 50%. PBS preservation shows loss of optical contrast within two hours, occurring predominantly in deep muscle and connective tissue. DMEM maintains tissue structure and optical scattering characteristics close to in vivo conditions up to 4 to 6 h after excision and best preserved tissue optical properties when compared to in vivo imaging.
Journal of Biomedical Optics 01/2005; 10(6):064033. · 2.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Optical coherence tomography (OCT) is an emerging medical imaging technology which can generate high resolution, cross-sectional images of tissue in situ and in real time, without the removal of tissue specimen. Although endoscopic OCT has been used successfully to identify certain pathologies in the gastrointestinal tract, the resolution of current endoscopic OCT systems has been limited to 10-15 um for clinical procedures. In this study, in vivo imaging of the gastrointestinal tract is demonstrated at a three-fold higher axial resolution (
[Show abstract][Hide abstract] ABSTRACT: High performance, short coherence length light sources with broad bandwidths and high output powers are critical for high-speed, ultrahigh resolution OCT imaging. We demonstrate a new, high performance light source for ultrahigh resolution OCT. Bandwidths of 140 nm at 1300 nm center wavelength with high output powers of 330 mW are generated by an all-fiber Raman light source based on a continuous-wave Yb-fiber laser-pumped microstructure fiber. The light source is compact, robust, turnkey and requires no optical alignment. In vivo, ultrahigh resolution, high-speed, time domain OCT imaging with <5 microm axial resolution is demonstrated.
[Show abstract][Hide abstract] ABSTRACT: A distally actuated, rotational-scanning micromotor endoscope catheter probe is demonstrated for ultrahigh-resolution in vivo endoscopic optical coherence tomography (OCT) imaging. The probe permits focus adjustment for visualization of tissue morphology at varying depths with improved transverse resolution compared with standard OCT imaging probes. The distal actuation avoids nonuniform scanning motion artifacts that are present with other probe designs and can permit a wider range of imaging speeds. Ultrahigh-resolution endoscopic imaging is demonstrated in a rabbit with <4-microm axial resolution by use of a femtosecond Cr:forsterite laser light source. The micromotor endoscope catheter probe promises to improve OCT imaging performance in future endoscopic imaging applications.
[Show abstract][Hide abstract] ABSTRACT: Ultrahigh resolution, real-time OCT is demonstrated with <5.4 um resolution in tissue at 1.4-1.7 um wavelengths using a high power continuum generated by a modelocked stretched-pulse Er-doped fiber laser and highly nonlinear fiber.
[Show abstract][Hide abstract] ABSTRACT: Current diagnostic imaging modalities of the thyroid gland cannot reliably distinguish benign from malignant lesions, primarily because of their inability to visualize microscopic structure. A high-resolution imaging technique capable of examining thyroid tissue architectural morphology in real time is needed. Optical coherence tomography (OCT) has been shown to achieve high resolutions approaching the cellular range (1-15 microm). The feasibility of optical coherence tomography for imaging thyroid tissue was explored ex vivo on the human thyroid gland.
High-resolution OCT was performed in real time at 2 to 4 frames per second on three postmortem and 15 surgically excised thyroid glands containing normal, hyperplastic, and neoplastic tissue. OCT images acquired were compared with those obtained using standard histopathologic methods.
The microstructure of the normal thyroid gland, including colloid-filled follicles as small as 15 microm and their supporting stroma, was clearly identified. OCT images of degenerative, hyperplastic, adenomatous, and malignant change within the thyroid gland were shown to correlate well with corresponding histopathologic findings.
The ability of OCT to image thyroid tissue microarchitecture makes it a potentially powerful technology that can be used to assess the thyroid gland at a resolution greater than currently available clinical imaging modalities.
Head & Neck 06/2004; 26(5):425-34. · 3.01 Impact Factor