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Ultrahigh resolution optical biopsy with endoscopic optical coherence tomography

Paul R. Herz, Yu Chen, Aaron D. Aguirre, James G. Fujimoto, Hiroshi Mashimo, Joseph Schmitt, Amanda Koski, John Goodnow, and Chris Petersen

Open Access Open Access

Abstract

Optical coherence tomography (OCT) is an emerging medical imaging technology that can generate high resolution, cross-sectional images of tissue in situ and in real time. 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 µm for in vivo studies. In this study, in vivo imaging of the rabbit gastrointestinal tract is demonstrated at a three-fold higher resolution (<5 µm), using a broadband Cr4+:Forsterite laser as the optical light source. Images acquired from the esophagus, trachea, and colon reveal high-resolution details of tissue architecture. Definitive correlation of architectural features in OCT images and histological sections is shown. The ability of ultrahigh resolution endoscopic OCT to image tissue morphology at an unprecedented resolution in vivo advances the development of OCT as a potential imaging tool for the early detection of neoplastic changes in biological tissue.

©2004 Optical Society of America

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Figures (9)
Fig. 1.
Fig. 1. Schematic of endoscopic OCT imaging system using a broadband Cr4+:Forsterite femtosecond laser light source.

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Fig. 2.
Fig. 2. (a) Optical bandwidth of the Cr4+:Forsterite light source at the input to the OCT system and transmitted through to the sample. The modelocked laser has high output power and excellent noise performance and enables a three-fold improvement in axial image resolution over standard endoscopic OCT systems using a superluminescent diode light source. (b) Measured axial resolution of 5 µm in air, corresponding to 3.7 µm resolution in tissue. (c) Plot of the logarithmic point spread function shows low sidelobes in the point spread function. The measured sensitivity was 102 dB with 10 mW of incident power on the sample.

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Fig. 3.
Fig. 3. (a) In vivo endoscopic OCT image of rabbit esophagus with (b) corresponding histology. Good correlation is seen between OCT and histology. The epithelium (e), lamina propria (lp), muscularis mucosa (mm), submucosa (sm), inner (im) and outer muscular (om) layers are visible on both the OCT image and histology.

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Fig. 4.
Fig. 4. (a) In vivo OCT image and (b) histology of rabbit esophagus and trachea viewed intraluminally from the esophagus. Tracheal hyaline cartilage (hc) between the tracheal mucosa and trachealis muscle is well defined. The image demonstrates the ability of the endoscopic OCT system to image deeply within the tissue. Trichrome stain was used to highlight cartilage and muscle layers.

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Fig. 5.
Fig. 5. In vivo image showing sequential OCT scans spanning the rabbit epiglottis to the inner esophagus. Ultrahigh resolution imaging capability is maintained over a large field allowing detailed discrimination of tissue structure. Architectural morphology of the proximal esophagus is well defined, as is the transition from the mouth to the esophagus at the epiglottis.

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Fig. 6.
Fig. 6. (a) Large-field scan of rabbit esophagus in vivo shows the transition region from the esophagus to the proximal stomach. There is excellent differentiation of esophageal versus gastric mucosa. Visible structures include epithelial folds (ef), esophageal mucosa (em), gastric transition (gt), and gastric mucosa (gm). (b) Corresponding histological cross section stained with hematoxylin and eosin.

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Fig. 7.
Fig. 7. (a) In vivo endoscopic OCT image of rabbit colon with (b) corresponding histology. Delineation of upper colonic mucosa (cm), muscular mucosa (mm), submucosa (sm), and muscularis externa (me) is possible. Enlarged images show the capability to visualize crypt structure. Tissue separations seen in the lower part of the histology images are due to a histology processing artifact.

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Fig. 8.
Fig. 8. (a) Rotational scanning images of rabbit esophagus and (b) proximal stomach in vivo. The esophageal lamina structure is well distinguished and high penetration is achieved. Epithelium (e), lamina propria (lp), inner muscularis (im), and outer (om) muscularis layers can be seen. Increased scattering with reduced image penetration is observed in the stomach, which is indicative of gastric mucosa architecture (gm).

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Fig. 9.
Fig. 9. (a) Rotational scan and (b) longitudinal pullback cross section of rabbit colon in vivo. Architectural detail is distinguished over the large scan field seen in the lower image. The location of the rotational image (a) in the pullback sequence is indicated by the solid line in (b). The cross-sectional imaging plane of the pullback scan is shown as a dashed line in the rotational image (a). The far right area in the pullback cross section is the animal rectum.

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