This PDF file contains the front matter associated with SPIE Proceedings Volume 8217, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and the Conference Committee listing.

A side-viewing, 2 mm diameter, surface magnifying chromoendoscopy (SMC)-optical coherence tomography (OCT) endoscope has been designed for simultaneous, non-destructive surface fluorescence visualization and cross-sectional imaging. We apply this endoscope to in vivo examination of mouse colon. A 30,000 element fiber bundle is combined with single mode fibers. The distal optics consist of a gradient-index lens and spacer to provide a magnification of 1 at a working distance of 1.58 mm in air, necessary to image the sample through a 0.23 mm thick outer glass envelope, and an aluminized right-angle prism fixed to the distal end of the GRIN lens assembly. The resulting 1:1 imaging system is capable of 3.9 μm lateral and 2.3 μm axial resolution in the OCT channel, and 125 lp/mm resolution across a 0.70 mm field of view in the SMC channel. The endoscope can perform high contrast crypt visualization, molecular imaging, and cross-sectional imaging of colon microstructure.

We report on the design and testing of a prototype widefield surface enhanced Raman scattering (SERS) imaging system based on a fiber optic bronchoscope using bandpass filters for Raman signal selection. The SERS contrast agents employed consist of gold nanoparticles encoded with a Raman-active dye and made specific for lung adenocarcinoma tissue through the use of an anti-epidermal growth factor receptor (EGFR) antibody. By exploiting the extremely narrow SERS spectral peaks we demonstrate a facile method of background fluorescence rejection that can be implemented at sub-video rates. The system has been tested on in-vivo tissues and performance metrics, including the maximum tissue penetration and minimum detectable nanoparticle quantity have been determined in a standardized fashion.

Multiphoton imaging methods are excellent for non-invasive imaging of living tissue without any need of additional contrast agents. The increasing demand for endoscopic techniques has forced the development of multiphoton endoscopes for imaging of areas with reduced accessibility like chronic wounds. Gradient index (GRIN) lenses can miniaturize the bulky distal focusing optics of conventional tomographs to a diameter of less than 1.4 mm and a numerical aperture (NA) of 0.8. We combined a high NA clinical multiphoton endoscope with existing multiphoton tomographs like the DermaInspect® and the MPTflex® to enable the examination of wound healing processes.

Optical coherence tomography (OCT) is a minimally-invasive imaging modality capable of tracking the development of individual colonic adenomas. As such, OCT can be used to evaluate the mechanisms and effectiveness of chemopreventive and chemotherapeutic agents in colorectal cancer models. The data presented here represent part of a larger study evaluating α-difluoromethylornithine (DFMO) and Sulindac as chemopreventive and chemotherapeutic agents using mice treated with the carcinogen azoxymethane (AOM). 27 A/J mice were included in the chemoprevention study, subdivided into four treatment groups (No Drug, DFMO, Sulindac, DFMO/Sulindac). 30 mm lateral images of each colon at eight different rotations were obtained at five different time points using a 2 mm diameter spectral domain OCT endoscopy system centered at 890 nm with 3.5 μm axial resolution in air and 5 μm lateral resolution. Images were visually analyzed to determine number and size of adenomas. Gross photos of the excised colons and histology provided gold standard confirmation of the final imaging time point. Preliminary results show that 100% of mice in the No Drug group developed adenomas over the course of the chemoprevention study. Incidence was reduced to 71.43% in mice given DFMO, 85.71% for Sulindac and 0% for DFMO/Sulindac. Discrete adenoma size did not vary significantly between experimental groups. Additional experiments are currently under way to verify these results and evaluate DFMO and Sulindac for chemotherapeutic applications.

A diode-pumped solid state laser is used to deliver excitation at λ_ex = 671 nm. The beam is expanded by a pair of relay lenses (f₁ = 30 and f₂ = 50 mm) to 3 mm diameter, filling the aperture of a fluid light cable that is coupled to a Hopkins II rigid endoscope. Near-infrared fluorescence images are collected by the endoscope and transmitted by another set of relay lenses onto a CCD detector that has dimensions of 8.7x6.9 mm² (1388x1040 pixels). A zoom lens system (F#1.6-16 aperture) with a tunable focal length (20-100 mm) magnifies the image to fill the dimensions of the CCD. A band pass filter allows fluorescence with spectral range λ_em = 696 to 736 nm to be collected. The system achieves a resolution of 9.8 μm and field-of-view of 3.6 mm at a distance of 2.5 mm between the distal end of the endoscope and the tissue. Images are collected at a rate of 10 frames per second. A filter wheel is incorporated into the handle of the instrument housing to rapidly switch between reflectance and fluorescence images. Cy5.5-labeled peptides were delivered through the 1 mm diameter instrument channel in the endoscope. Near-infrared fluorescence images demonstrated specific peptide binding to spontaneous adenomas that developed beginning at 2 months of age in a genetically-engineered mouse with mutation of one allele in the APC gene. This integrated methodology represents a powerful tool that can achieve real time detection of disease in the colon and other hollow organs.

We demonstrate a multi-spectral scanning fiber endoscope (SFE) that collects fluorescence images in vivo from three target peptides that bind specifically to murine colonic adenomas. This ultrathin endoscope was demonstrated in a genetically engineered mouse model of spontaneous colorectal adenomas based on somatic Apc (adenomatous polyposis coli) gene inactivation. The SFE delivers excitation at 440, 532, 635 nm with <2 mW per channel. The target 7-mer peptides were conjugated to visible organic dyes, including 7-Diethylaminocoumarin-3-carboxylic acid (DEAC) (λ_ex=432 nm, λ_em=472 nm), 5-Carboxytetramethylrhodamine (5-TAMRA) (λ_ex=535 nm, λ_em=568 nm), and CF-633 (λ_ex=633 nm, λ_em=650 nm). Target peptides were first validated using techniques of pfu counting, flow cytometry and previously established methods of fluorescence endoscopy. Peptides were applied individually or in combination and detected with fluorescence imaging. The ability to image multiple channels of fluorescence concurrently was successful for all three channels in vitro, while two channels were resolved simultaneously in vivo. Selective binding of the peptide was evident to adenomas and not to adjacent normal-appearing mucosa. Multispectral wide-field fluorescence detection using the SFE is achievable, and this technology has potential to advance early cancer detection and image-guided therapy in human patients by simultaneously visualizing multiple over expressed molecular targets unique to dysplasia.

Optical Coherence Tomography catheters comprise a transparent tube which can act as a negative cylindrical lens and introduce astigmatism which will lead to a decrease in transverse resolution and image contrast. In this report, we numerically analyzed the astigmatism for standard catheter designs applicable to esophageal and coronary imaging. In order to maintain image quality, generally the beam can be refocused by a curved interface. To handle a situation involving high-index flush media, another method based on matching refractive indices is described and shown to successfully restore a round beam.

Recent evidence suggests that epithelial ovarian cancer may originate in the fimbriated end of the fallopian tube¹. Unlike many other cancers, poor access to the ovary and fallopian tubes has limited the ability to study the progression of this deadly disease and to diagnosis it during the early stage when it is most amenable to therapy. We have previously reported on a rigid confocal microlaparoscope system that is currently undergoing a clinical trial to image the epithelial surface of the ovary². In order to gain in vivo access to the fallopian tubes we have developed a new confocal microlaparoscope with an articulating distal tip. The new instrument builds upon the technology developed for the existing confocal microlaparoscope. It has an ergonomic handle fabricated by a rapid prototyping printer. While maintaining compatibility with a 5 mm trocar, the articulating distal tip of the instrument consists of a 2.2 mm diameter bare fiber bundle catheter with automated dye delivery for fluorescence imaging. This small and flexible catheter design should enable the confocal microlaparoscope to image early stage ovarian cancer arising inside the fallopian tube. Early ex vivo mages of human fallopian tube and in vivo imaging results from recent open surgeries using the rigid confocal microlaparoscope system are presented. Ex vivo images from animal models using the new articulating bare fiber system are also presented. These high quality images collected by the new flexible system are similar in quality to those obtained from the epithelial surface of ovaries with the rigid clinical confocal microlaparoscope.