Label-Free In Vivo Imaging of Corneal Lymphatic Vessels Using Microscopic Optical Coherence Tomography.

Purpose: Corneal neovascularization, in particular lymphangiogenesis, is a limiting factor in corneal transplant survival. Novel treatment approaches focus on (selective) inhibition and regression of lymphatic vessels. Imaging clinically invisible corneal lymphatic vessels is a prerequisite for these strategies. Using a murine model, this study investigates whether corneal lymphatic vessels can be imaged using microscopic optical coherence tomography (mOCT). Methods: Corneal neovascularization was induced by intrastromal placement of 11.0 nylon sutures in one eye of BALB/c mice. After 2 weeks, cross-sectional images and volumes of the corneas with a 0.5 mm lateral and axial field of view were acquired using a custom-built mOCT system enabling a resolution of 1 µm at a B-scan rate of 165/s. Three of the six animals received an additional intrastromal injection of India ink 24 hours before the measurement to stain the corneal lymphatic system in vivo. Immunohistochemistry using CD31 and LYVE-1 was used to validate the mOCT findings. Results: Using mOCT, lymphatic vessels were visible as dark vessel-like structures with the lumen lacking a hyperreflective wall and mostly lacking cells. However, individual, slowly moving particles, which most likely are immune cells, occasionally could be observed inside the lumen. In lymphatic vessels of ink-stained corneas, hyperreflection and shadowing underneath was observed. Ink-filled lymphatic vessels were colocalized in consecutive corneal flat mounts of the same specimen. Conclusions: Corneal lymphatic vessels can be imaged using mOCT. This novel approach opens new options for noninvasive clinical imaging of corneal lymphatic vessels for diagnostic and therapeutic indications.

Intraoperative assessment of laryngeal pathologies with optical coherence tomography integrated into a surgical microscope.

OBJECTIVE: Endoscopic examination followed by tissue biopsy is the gold standard in the evaluation of lesions of the upper aerodigestive tract. However, it can be difficult to distinguish between healthy mucosa, dysplasia, and invasive carcinoma. Optical coherence tomography (OCT) is a non-invasive technique which acquires high-resolution, cross-sectional images of tissue in vivo. Integrated into a surgical microscope, it allows the intraoperative evaluation of lesions simultaneously with microscopic visualization. STUDY DESIGN: In a prospective case series, we evaluated the use of OCT integrated into a surgical microscope during microlaryngoscopy to help differentiating various laryngeal pathologies. METHODS: 33 patients with laryngeal pathologies were examined with an OCT- microscope (OPMedT iOCT-camera, HS Hi-R 1000G-microscope, Haag-Streit Surgical GmbH, Wedel, Germany) during microlaryngoscopy. The suspected intraoperative diagnoses were compared to the histopathological reports of subsequent tissue biopsies. RESULTS: Hands-free non-contact OCT revealed high-resolution images of the larynx with a varying penetration depth of up to 1.2 mm and an average of 0.6 mm. Picture quality was variable. OCT showed disorders of horizontal tissue layering in dysplasias with a disruption of the basement membrane in carcinomas. When comparing the suspected diagnosis during OCT-supported microlaryngoscopy with histology, 79% of the laryngeal lesions could be correctly identified. Premalignant lesions were difficult to diagnose and falsely classified as carcinoma. CONCLUSION: OCT integrated into a surgical microscope seems to be a promising adjunct tool to discriminate pathologies of the upper aerodigestive tract intraoperatively. However, picture quality and penetration depth were variable. Although premalignant lesions were difficult to diagnose, the system proved overall helpful for the intraoperative discrimination of benign and malignant tumors. Further studies will be necessary to define its value in the future. Lasers Surg. Med. 49:490-497, 2017. © 2017 Wiley Periodicals, Inc.

Detection of cervical intraepithelial neoplasia by using optical coherence tomography in combination with microscopy.

Optical coherence tomography (OCT) is a noninvasive high-resolution imaging technique that permits the detection of cancerous and precancerous lesions of the uterine cervix. The purpose of this study was to evaluate a new system that integrates an OCT device into a microscope. OCT images were taken from loop electrosurgical excision procedure (LEEP) specimens under microscopic guidance. The images were blinded with respect to their origin within the microscopic image and analyzed independently by two investigators using initially defined criteria and later compared to the corresponding histology. Sensitivity and specificity were calculated with respect to the correct identification of high-grade squamous intraepithelial lesions (HSIL). The interinvestigator agreement was assessed by using Cohen's kappa statistics. About 160 OCT images were obtained from 20 LEEP specimens. Sixty randomly chosen images were used to define reproducible criteria for evaluation. The assessment of the remaining 100 images showed a sensitivity of 88% (second investigator 84%) and a specificity of 69% (65%) in detecting HSIL. Surgical microscopy-guided OCT appears to be a promising technique for immediate assessment of microanatomical changes. In the gynecological setting, the combination of OCT with a colposcope may improve the detection of high-grade squamous intraepithelial lesions.

Microanatomy of the tympanic membrane in chronic myringitis obtained with optical coherence tomography.

A microscope-based optical coherence tomography (OCT) device was used to assess the microanatomy of the tympanic membrane in patients with chronic myringitis. A prospective study was designed for this purpose. OCT measurements of the tympanic membrane were done on 11 patients with myringitis with a microscope-based spectral domain OCT system. The in vivo findings were compared with those findings of a control group consisting of 36 patients with retraction pockets or atrophic tympanic membranes (n = 13), myringosclerosis (n = 12) and perforations (n = 11). In active chronic myringitis, the thickness of the tympanic membrane is increased compared to healthy membranes and to other pathological conditions of the tympanic membrane. Consistent changes of the microanatomy of the tympanic membrane were found in chronic myringitis with OCT. Serial OCT measurements revealed no biofilm suspicious findings in all patients with active chronic myringitis. Intraoperative and in vivo OCT measurements may help to detect microanatomical changes of the tympanic membrane in chronic myringitis and in other conditions of the tympanic membrane.

Future prospects: assessment of intraoperative optical coherence tomography in ab interno glaucoma surgery.

PURPOSE: Ab interno glaucoma surgery, such as trabecular aspiration or ab interno trabeculotomy, has been introduced as an alternative, non-filtering procedure to lower intraocular pressure in glaucoma patients. The purpose of the present study was to assess the feasibility of intraoperative spectral domain optical coherence tomography (iOCT) for improving ab interno glaucoma surgery. MATERIALS AND METHODS: Intraoperative optical coherence tomography was performed to visualize the anterior chamber angle, the aspiration canula during trabecular aspiration, and the trabectome device during ab interno trabeculotomy. A commercially available 840-nm iOCT was used in a clinical setting, a 1310-nm spectral-domain OCT in an experimental setting. RESULTS: In the clinical setting using the 840-nm iOCT, visualization of the aspiration canula and the trabectome device was possible, however, with rather limited visibility of the chamber angle and the trabecular meshwork. In the experimental setting, the 1310-nm OCT enabled excellent visualization of the chamber angle and detailed evaluation of the instrument-trabecular meshwork relation. CONCLUSIONS: Intraoperative use of optical coherence tomography may be helpful for ab interno glaucoma surgery. Adjustments of the commercially available iOCT, including longer wavelengths and oblique scanning options, are necessary for the clinical use.

Optimising deep anterior lamellar keratoplasty (DALK) using intraoperative online optical coherence tomography (iOCT).

BACKGROUND/AIMS: To describe the use of intraoperative online optical coherence tomography (iOCT) for improving deep anterior lamellar keratoplasty (DALK) surgery. METHODS: Retrospective case series of 6 eyes of 6 male patients with keratokonus, corneal dystrophy or herpetic stromal scars undergoing DALK were investigated using intraoperative optical coherence tomography and postsurgical image/video analysis. Main outcome measures were: visibility of surgical steps, especially, assessment of placement depth of injection needle, preparation of bare Descemet's membrane and drainage of interface fluid. RESULTS: iOCT enables real-time visualisation of all surgical steps of DALK procedure in all patients. Placement of air injection needle above Descemet's membrane was reliably monitored as was presence of bare Descemet's membrane and potential interface fluid. CONCLUSIONS: iOCT assists with visualisation of injection needle placement and with assessment of bare Descemet's membrane as well as interface fluid during the DALK procedure. Overall iOCT may be a helpful device that supports surgeons in all steps of DALK procedure.

Optimizing descemet membrane endothelial keratoplasty using intraoperative optical coherence tomography.

IMPORTANCE: Descemet membrane endothelial keratoplasty (DMEK) is a challenging procedure for the surgeon, particularly because of deficient visibility of the delicate tissue due to the natural en face view through the operating microscope. A cross-sectional view would greatly enhance intraoperative overview and enable the surgeon to better control the procedure. OBJECTIVE: To retrospectively analyze the use of intraoperative optical coherence tomography (iOCT) for improving the safety of DMEK. DESIGN: Intraoperative OCT during DMEK was performed in 26 eyes of 26 patients. We retrospectively analyzed imaging and video data. SETTING: Department of Ophthalmology, University of Cologne. PARTICIPANTS: Seven men and 19 women aged 39 to 93 years with corneal endothelial dysfunction undergoing DMEK. EXPOSURE: Descemet membrane endothelial keratoplasty. MAIN OUTCOMES AND MEASURES: Visibility of surgical steps, overall duration of DMEK, overall time for complete intraoperative air filling of the anterior chamber, and correlation between donor age and Descemet rolling behavior. RESULTS Intraoperative OCT enables visualization of all steps of the DMEK procedure. Overall mean (SD) duration of the DMEK procedure was 25.7 (6.9) minutes when using iOCT. Overall mean (SD) complete intraoperative anterior chamber air-filling time was 236 (108) seconds in contrast to 60 to 90 minutes for standard air-filling time. Descemet membrane rolling behavior showed significant inverse correlation between donor age (range, 39-93 years) and the extent of rolling (R2 = 0.5 [P = .006]). CONCLUSIONS AND RELEVANCE: Intraoperative OCT enhances the visibility of graft orientation and unfolding, thereby improving safety of the DMEK procedure. Overall, iOCT is a helpful device that may support surgeons in all steps of DMEK procedures.

[Diagnosis of the deep partial-thickness burn wound of Skh-1 mouse with Optical Coherence Tomography].

OBJECTIVE: To evaluate the application value of Optical Coherence Tomography (OCT) in the diagnosis of the depth of burn wound. METHODS: Deep partial-thickness scald models of Skh-1 mice were reproduced using self-made steam scald appliance. The scald wounds were scanned with OCT 3 hours, or 3 and 8 days after injury respectively. Scanned wound tissue was harvested for histological examination right after each episode of OCT imaging. Normal skin of mice was scanned and examined with the above-mentioned methods at the same time. RESULTS: Compared with those of the normal skin, collagen in the dermis was denatured after steam scald, and it was imaged as vanishing or reduction in birefringence in OCT detection. The structure change intensity was related to the pathological process of the wounds and consistent with the corresponding histological results. CONCLUSIONS: OCT is a noninvasive technique. It can be used to diagnose the depth of burn wound in real time.

Optical coherence tomography allows for the reliable identification of laryngeal epithelial dysplasia and for precise biopsy: a clinicopathological study of 61 patients undergoing microlaryngoscopy.

OBJECTIVES/HYPOTHESIS: A newly developed microscope-based spectral-domain optical coherence tomography (SD-OCT) device and an endoscope-based time-domain OCT (TD-OCT) were used to assess the inter-rater reliability, sensitivity, specificity, and accuracy of benign and dysplastic laryngeal epithelial lesions. STUDY DESIGN: Prospective study. METHODS: OCT during microlaryngoscopy was done on 35 patients with an endoscope-based TD-OCT, and on 26 patients by an SD-OCT system integrated into an operating microscope. Biopsies were taken from microscopically suspicious lesions allowing comparative study of OCT images and histology. RESULTS: Thickness of the epithelium was seen to be the main criterion for degree of dysplasia. The inter-rater reliability for two observers was found to be kappa = 0.74 (P <.001) for OCT. OCT provided test outcomes for differentiation between benign laryngeal lesions and dysplasia/CIS with sensitivity of 88%, specificity of 89%, PPV of 85%, NPV of 91%, and predictive accuracy of 88%. However, because of the limited penetration depth of the laser light primarily in hyperkeratotic lesions (thickness above 1.5 mm), the basal cell layer was no longer visible, precluding reliable assessment of such lesions. CONCLUSIONS: OCT allows for a fairly accurate assessment of benign and dysplastic laryngeal epithelial lesion and greatly facilitates the taking of precise biopsies. Laryngoscope, 2010.

Optical coherence tomography with online visualization of more than seven rendered volumes per second.

Nearly real-time visualization of 3-D volumes is crucial for the use of optical coherence tomography (OCT) during microsurgery. With an ultrahigh speed spectral domain OCT coupled to a surgical microscope, on-line display of 7.2 rendered volumes at 87 megapixels per second is demonstrated. Calculating the A-scans from the spectra is done on a quad-core personal computer (PC), while dedicated software for the 3-D rendering is executed on a high performance video card. Imaging speed is practically only limited by the readout of the camera. First experiments show the feasibility of real-time 3-D OCT for guided interventions.

Optical coherence tomography for process control of laser micromachining.

In situ surface imaging for nondestructive evaluation (NDE) by optical coherence tomography (OCT) before, during, and after ablative laser processing is presented. Furthermore, it is shown that the ability of in situ characterization is beneficial for samples such as optical fibers, which are difficult to handle in the standard analysis. Surface images taken by the OCT are compared with these common analysis tools such as scanning electron microscopy (SEM), reflected-light, and confocal microscopy. An axial resolution of approximately 126 nm for surface detection and a lateral resolution <2.5 microm are obtained and the potential of the setup to imaging structures with high aspect ratio is demonstrated.

Optical coherence tomography as an orientation guide in cochlear implant surgery?

CONCLUSION: With optical coherence tomography (OCT) it is basically possible to reveal parts of the cochlear morphology without opening its enveloping membranes. Thus, it may serve as a helpful guide for the surgeon to localize the scala tympani precisely before opening the fluid-filled inner ear to insert the electrode array. OBJECTIVE: To improve anatomical orientation in cochlear implant surgery before definitively opening the fluid-filled inner ear. The question was whether a new imaging technique, OCT, might provide information about the site of the underlying inner ear structures (scala tympani, scala vestibuli) and could, consequently, guide the surgeon towards the scala tympani. MATERIALS AND METHODS: In a preliminary study, OCT was carried out on human temporal bone preparations, in which a cochleostomy ('fenestration') was performed leaving the endosteum and the fluid-filled inner ear intact. OCT was applied via a prototype of a specially equipped operating microscope. The mode of OCT used in this context was spectral-domain (SD)-OCT. RESULTS: On scans, which can be read analogous to B-mode sonography, OCT provides information about structures on the inner surface of the partly exposed but still intact membranous cochlear lining - such as scala tympani or scala vestibuli.