We have designed a multimodality system that combines optical coherence tomography (OCT) and laser-induced fluorescence (LIF) in a 2.0-mm-diameter endoscopic package. OCT provides approximately 18-microm resolution cross-sectional structural information over a 6-mm field. LIF spectra are collected sequentially at submillimeter resolution across the same field and provide histochemical information about the tissue. We present the use of a rod prism to reduce the asymmetry in the OCT beam caused by a cylindrical window. The endoscope has been applied to investigate mouse colon cancer in vivo.

Frequency domain optical coherence tomography (FD-OCT) allows interferometer topologies with simplified system construction and handling. Problems of dispersion and polarization matching between the sample and reference arms, as well as beamsplitter spectral non-uniformity, are mitigated when the interferometer is wholly contained in the endoscope tip. A common path set-up, using a reference reflection originating from the inside surface of the glass envelope at the distal end of the endoscope, and an alternative approach with more efficient collection of the reference light using a novel beamsplitter design have been developed. High speed (20,000 A-lines/s) ultrahigh axial resolution (2.4 mum) tomograms of mouse colon have been acquired using a 2 mm outer diameter endoscope in vivo. The FD-OCT system uses a compact mode-locked Ti:Al(2)O(3) laser emitting a broad spectrum (160 nm full-width-half-maximum) centered at 800 nm in combination with a CCD based, spectrally sensitive detector.

: Recent substantial developments in light source and detector technology have initiated a paradigm shift in retinal optical coherence tomography (OCT) performance. Broad bandwidth light sources in the 800 nm and 1060 nm wavelength region enable axial OCT resolutions of 2-3 mum and 5-7 mum, respectively. Novel high speed silicon based CMOS cameras at 800 nm and InGaAs based CCD cameras in combination with frequency domain OCT technology enable data acquisition speeds of up to 47,000 A-scans/s at 1060 nm and up to 312,500 A-scans/s at 800 nm. Combining ultrahigh axial resolution, ultrahigh speed OCT at 800 nm with pancorrected adaptive optics allows volumetric in vivo cellular resolution retinal imaging. Commercially available three-dimensional (3D) retinal OCT at 800 nm (20,000 A-scans/s, 6 mum axial resolution) is compared to ultrahigh speed 3D retinal imaging at 800 nm (160,000 A-scans/s, 2-3 mum axial resolution), high speed 3D choroidal imaging at 1060 nm (47,000 Ascan/ second, 6-7 mum axial resolution) and cellular resolution retinal imaging at 800 nm using adaptive optics OCT at 160,000 A-scans/second with isotropic resolution of ~2 mum. Analysis of the performance of these four imaging modalities applied in normal and pathologic eyes focusing on motion artifact free volumetric retinal imaging and revealing novel, complementary morphological information due to enhanced resolution, speed and penetration is presented.

These preliminary data indicate endoscopic OCT-LIF has the potential to identify colorectal adenomas in murine models.

Endoscopic ultrahigh-resolution optical coherence tomography (OCT) enables collection of minimally invasive cross-sectional images in vivo, which may be used to facilitate rapid development of reliable mouse models of colon disease as well as assess chemopreventive and therapeutic agents. The small physical scale of mouse colon makes light penetration less problematic than in other tissues and high resolution acutely necessary. In our 2-mm diameter endoscopic time domain OCT system, isotropic ultrahigh-resolution is supported by a center wavelength of 800 nm and full-width-at-half-maximum bandwidth of 150 nm (mode-locked titanium:sapphire laser) combined with 1:1 conjugate imaging of a small core fiber. A pair of KZFSN5/SFPL53 doublets provides excellent color correction to support wide bandwidth throughout the imaging depth. A slight deviation from normal beam exit angle suppresses collection of the strong back reflection at the exit window surface. Our system achieves axial resolution of 3.2 microm in air and 4.4-microm lateral spot diameter with 101-dB sensitivity. Microscopic features too small to see in mouse tissue with conventional resolution systems, including colonic crypts, are clearly resolved. Resolution near the cellular level is potentially capable of identifying abnormal crypt formation and dysplastic cellular organization.

KEy WoRDSendoscopic imaging, imaging of tumor progression, gastrointestinal cancer, colorectal adenoma, gastrointestinal intraepithelial neoplasia, animal models for carcinogenesis, azoxymethane treated mouse model of colorectal neoplasm Research PaperSerial Endoscopy in Azoxymethane Treated Mice Using Ultra-High Resolution Optical Coherence Tomography ABSTRAcT Purpose: Optical coherence tomography (OCT) is a minimally invasive, depth-resolved imaging tool that can be implemented in a small diameter endoscope for imaging mouse models of colorectal cancer (CRC). In this study, we utilized ultrahigh resolution (UHR) OCT to serially image the lower colon of azoxymethane (AOM) treated A/J mouse models of CRC in order to monitor the progression of neoplastic transformations and determine if OCT is capable of identifying early disease.Experimental Design: Thirteen AOM treated A/J and two control A/J mice were surveyed at four timepoints (8, 14, 22 and 26 weeks post AOM treatment) using a 2.0 mm diameter UHR OCT endoscopic system with 3.2 mm axial and 4.4 mm lateral resolution. Histological samples obtained at the final timepoint served as the diagnostic reference. A blinded expert panel of mouse colon pathologists provided diagnoses from the OCT images based on criteria developed from a separate training set of OCT images. Panel results were compared to histological diagnoses assigned by a blinded pathologist.Results: At the final imaging timepoint, 95% of adenomas and 23% of gastrointestinal neoplasias (38% protruding GINs and 9% non-protruding GINs) were correctly diagnosed. The panel identified 68% of disease foci (95% adenoma, 76% protruding GINs and 13% non-protruding GINs). Over the OCT imaging timepoints, disease progression followed a typical succession, with normal or GIN preceding adenoma.Conclusions: Endoscopic UHR OCT enabled accurate diagnosis of adenomas, identification of protruding GIN and non-destructive visualization of CRC progression, providing a tool for cancer research in animal models.

Optical coherence tomography (OCT), laser-induced fluorescence (LIF), and laser-scanning confocal microscopy (LSCM) were used for the task of multimodal study of healthy and adenomatous mouse colon. The results from each modality were compared with histology, which served as the gold standard. The Apc(Min/+) genetic mouse model of colon cancer was compared with wild-type mice. In addition, a special diet was used for the task of studying the origins of a 680 nm autofluorescent signal that was previously observed in colon. The study found close agreement among each of the modalities and with histology. All four modalities were capable of identifying diseased tissue accurately. The OCT and LSCM images provided complementary structural information about the tissue, while the autofluorescence signal measured by LIF and LSCM provided biochemical information. OCT and LIF were performed in vivo and nondestructively, while the LSCM and histology required extraction of the tissue. The magnitude of the 680 nm signal correlates with chlorophyll content in the mouse diet, suggesting that the autofluorescent compound is a dietary metabolite.

We develop a dual-modality device that combines the anatomical imaging capabilities of optical coherence tomography (OCT) with the functional capabilities of laser-induced fluorescence (LIF) spectroscopy. OCT provides cross-sectional images of tissue structure to a depth of up to 2 mm with approximately 10-microm resolution. LIF spectroscopy provides histochemical information in the form of emission spectra from a given tissue location. The OCT subsystem utilizes a superluminescent diode with a center wavelength of 1300 nm, whereas a helium cadmium laser provides the LIF excitation source at wavelengths of 325 and 442 nm. Preliminary data are obtained on eight postmortem aorta samples, each 10 mm in length. OCT images and LIF spectra give complementary information from normal and atherosclerotic portions of aorta wall. OCT images show structures such as intima, media, internal elastic lamina, and fibrotic regions. Emission spectra ratios of 520/490 (325-nm excitation) and 595/635 (442-nm excitation) could be used to identify normal and plaque regions with 97 and 91% correct classification rates, respectively. With miniaturization of the delivery probe and improvements in system speed, this dual-modality device could provide a valuable tool for identification and characterization of atherosclerotic plaques.