Ординатура / Офтальмология / Английские материалы / Handbook of Optical Coherence Tomography_Bouma, Tearney_2002
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tically assessing microsurgical procedures using an OCT microscope delivery system [54]. Figure 17 shows OCT images of an arterial anastomosis of a rabbit inguinal artery, demonstrating the ability of OCT to assess internal structure and luminal patency. An artery segment was bisected cross-sectionally and then reanastomosed. For precise registration of the OCT images, the specimen was positioned on a computer-controlled, motorized translation stage in an OCT microscope. The specimen was also digitally imaged with a CCCD camera. Cross-sectional OCT images of a 1 mm diameter rabbit inguinal artery are shown. Figures 17A–D were acquired transversely at different positions through the anastomosis. The images of the ends of the artery clearly show arterial morphology corresponding to the intimal, medial, and adventitial layers of the elastic artery. The image from the site of the anastomosis shows that the lumen was obstructed by a tissue flap. By assembling a series of cross-sectional twodimensional images, a three-dimensional dataset was produced. Arbitrary planes can be selected and sections displayed.
Figure 17 Microsurgical imaging. OCT images of an anastomosis in a rabbit artery as an example of microsurgical imaging. The 1 mm diameter rabbit artery was anastomosed with a continuous suture as seen in the digital image (D). The labels indicate the planes from which corresponding cross-sectional OCT images were acquired. (A,D) Opposite ends of the anastomosis show the multilayered structure of the artery with a patent lumen. (B) Partially obstructed lumen and the presence of a thrombogenic flap. (C) Fully obstructed portion of the anastomosis site. (E) Three-dimensional reconstruction of artery. (G) Virtual longitudinal section view constructed from three-dimensional image data sets shows the obstruction (o). OCT microscopy could be a powerful tool for many microsurgical procedures. (From Ref. 54.)
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Because OCT imaging can be performed at high speeds, in real time, it can be integrated directly with surgery. The feasibility of using high speed OCT imaging to guide the placement and image the dynamics of surgical laser ablation has been investigated [55]. Figure 18 shows an example of real-time OCT imaging during laser ablation of a bovine muscle specimen. The laser exposure was 2 W from the argon laser. At 0.5 s changes in optical properties of the tissue are observed due to heating, and explosive ablation begins at 2.25 s. These studies were performed using a commercially available low coherence light source (AFC Technologies Inc., Hull, Quebec, Canada) with a wavelength at 1:3 m and an axial resolution of 18 m. Using 5 mW of incident power on the specimen, a signal-to-noise ratio of 115 dB was achieved. The acquisition rate was 8 frames per second for a 250 transverse pixel image. Optical coherence tomography should improve intraoperative diagnostics by providing high resolution, subsurface, cross-sectional imaging in real time. A comprehensive discussion of OCT applications for the guidance of intervention is presented in Chapter 23.
1.12HIGH RESOLUTION CELLULAR LEVEL OCT IMAGING
The development of high resolution OCT is also an important area of active research. Increasing resolution to the cellular and subcellular levels is important
Figure 18 Real-time OCT imaging of laser ablation showing a series of OCT images taken at 8 frames per second during argon laser exposure of beef muscle. The exposure was 1 W with a 0.8 mm spot diameter on tissue. The times indicated are in seconds after exposure is initiated. At 2.25 s, the formation of a blister at the surface is observed. At 2.5 s, the blister explodes and a crater develops (5 s). These images show the ability of OCT to perform high speed, real-time imaging during interventional procedures. (From Ref. 55.)
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for many applications, including the diagnosis of early neoplasias. As discussed previously, the axial resolution of OCT is determined by the coherence length of the light source used for imaging. Light sources for OCT imaging should have a short coherence length or broad bandwidth but also must have a single spatial mode to enable interferometry. Since the signal-to-noise ratio depends on the incident power, light sources with average powers of several milliwatts are typically necessary to achieve real-time imaging One approach for achieving high resolution is to use short-pulse femtosecond solid-state lasers as light sources [22,43,44,112].
High resolution OCT imaging has been demonstrated in vivo in developmental biology specimens [22,112]. Figure 19 shows an example of high resolution OCT images of a Xenopus laevis (African frog) tadpole. The OCT system in this study was a KLM femtosecond Ti : sapphire laser that emits sub-two-cycle pulses, corresponding to bandwidths of up to 350 nm (FWHM) around 800 nm. The ultrahigh resolution OCT system supported bandwidths up to 260 nm (FWHM), achieving a 1:5 m longitudinal resolution in free space of 1 m in tissue. To overcome the depth-of- field limitations associated with the high transverse resolution, a zone focus and image fusion technique was used. The figure was constructed by fusing multiple separate images recorded with different focal depths of the optics similar to C- mode scanning in ultrasound. OCT can image nuclear and intracellular morphology as well as identify cells in different stages of mitosis and visualize mitotic activity.
Figure 19 High resolution OCT images of a Xenopus laevis (African frog) tadpole in vivo performed with 1 m axial and 3 m transverse resolution. An image fusion technique is used to overcome depth-of-field limitations associated with the high transverse resolution. Individual cells and nuclei are clearly visible. Cellular level imaging has important implications for a variety of OCT applications. (See Refs. 112 and 22.)
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The ability to image subcellular structure can be an important tool for studying mitotic activity and cell migration in developmental biology. The extension of these results to human cells has important implications. Because differentiated human cells are smaller than developing cells, additional improvements in resolution and performance are necessary. In ophthalmology, improving the resolution should improve morphometric measurements such as retinal thickness and retinal nerve fiber layer thickness, which are relevant for the detection of macular edema and glaucoma. High resolution imaging also has implications for diagnosis of neoplasia. Standard OCT image resolutions enable the imaging of architectural morphology on the 10– 15 m scale and can identify many early neoplastic changes. The extension of imaging to the cellular and subcellular levels would not only enhance the spectrum of early neoplasias and dysplasias that could be imaged but also improve sensitivity and specificity.
1.13SUMMARY
Optical coherence tomography is a fundamentally new imaging modality with rapidly emerging applications spanning a range of fields. OCT can perform micro- meter-scale imaging of internal microstructure in materials and biological tissues in situ and in real time. For medical imaging applications, OCT can function as a type of optical biopsy to obtain microstructural information with resolution approaching that of conventional histopathology. Image information is available immediately without the need for excise and histological processing of a specimen. The development of high speed OCT technology now permits real-time imaging with high pixel densities. A wide range of OCT imaging platforms and probes have been developed, including ophthalmoscopes, microscopes, laparoscopes, hand-held probes, and miniature, flexible catheter-endoscopes. These imaging probes can be used either separately or in conjunction with other medical imaging instruments such as endoscopes and bronchoscopes and can permit internal body imaging in a wide range of organ systems. There are numerous research applications of this technology in a broad range of fields as well as continuing development of the technology itself. More research remains to be done, and numerous clinical studies must be performed to identify the clinical situations in which OCT can play a role. However, the unique capabilities of OCT imaging suggest that it could have a significant impact on fundamental research as well as on health care.
ACKNOWLEDGMENTS
Our research would not have been possible without the long-term collaboration and support of a talented multidisciplinary research team. The invaluable contributions of Dr. Mark Brezinski of the Massachusetts General Hospital and Harvard Medical School, Dr. Joel Schuman and Dr. Carmen Puliafito of the New England Eye Center and Tufts University School of Medicine, and Eric Swanson of Coherent Diagnostic Technology are greatly appreciated. We acknowledge the contributions of visiting scientists, including Dr. Wolfgang Drexler of the University of Vienna, Dr. Juergen Herrmann of the University of Erlangen, and Stephan Kubasiak of the Laser Medical Center of Lubeck. Present and former group members—postdoctoral associates, MD/PhD students, and PhD students, including Dr. Stephen Boppart,
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Dr. Brett Bouma, Ravi Ghanta, Dr. David Huang, Dr. Michael Hee, Christine Jesser, Dr. Xingde Li, Constantinos Pitris, and Dr. Gary Tearney—have made invaluable contributions. This research has been supported in part by the National Institutes of Health, contracts NIH-1-RO1-CA75289-02 and NIH-1- RO1-EY11289-13, the Medical Free Electron Laser Program, Office of Naval Research contract N000014-97-1-1066, and the Air Force Office of Scientific Research, contract F4920-98-1-0139.
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