In vivo high resolution human corneal imaging using full-field optical coherence tomography.

We present the first full-field optical coherence tomography (FFOCT) device capable of in vivo imaging of the human cornea. We obtained images of the epithelial structures, Bowman's layer, sub-basal nerve plexus (SNP), anterior and posterior stromal keratocytes, stromal nerves, Descemet's membrane and endothelial cells with visible nuclei. Images were acquired with a high lateral resolution of 1.7 µm and relatively large field-of-view of 1.26 mm x 1.26 mm - a combination, which, to the best of our knowledge, has not been possible with other in vivo human eye imaging methods. The latter together with a contactless operation, make FFOCT a promising candidate for becoming a new tool in ophthalmic diagnostics.

Multimodal imaging of a humanized orthotopic model of hepatocellular carcinoma in immunodeficient mice.

The development of multimodal strategies for the treatment of hepatocellular carcinoma requires tractable animal models allowing for advanced in vivo imaging. Here, we characterize an orthotopic hepatocellular carcinoma model based on the injection of luciferase-expressing human hepatoma Huh-7 (Huh-7-Luc) cells in immunodeficient mice. Luciferase allows for an easy repeated monitoring of tumor growth by in vivo bioluminescence. The intrahepatic injection was more efficient than intrasplenic or intraportal injection in terms of survival, rate of orthotopic engraftment, and easiness. A positive correlation between luciferase activity and tumor size, evaluated by Magnetic Resonance Imaging, allowed to define the endpoint value for animal experimentation with this model. Response to standard of care, sorafenib or doxorubicin, were similar to those previously reported in the literature, with however a strong toxicity of doxorubicin. Tumor vascularization was visible by histology seven days after Huh-7-Luc transplantation and robustly developed at day 14 and day 21. The model was used to explore different imaging modalities, including microtomography, probe-based confocal laser endomicroscopy, full-field optical coherence tomography, and ultrasound imaging. Tumor engraftment was similar after echo-guided intrahepatic injection as after laparotomy. Collectively, this orthotopic hepatocellular carcinoma model enables the in vivo evaluation of chemotherapeutic and surgical approaches using multimodal imaging.

Assessment of Sentinel Node Biopsies With Full-Field Optical Coherence Tomography.

Current techniques for the intraoperative analysis of sentinel lymph nodes during breast cancer surgery present drawbacks such as time and tissue consumption. Full-field optical coherence tomography is a novel noninvasive, high-resolution, fast imaging technique. This study investigated the use of full-field optical coherence tomography as an alternative technique for the intraoperative analysis of sentinel lymph nodes. Seventy-one axillary lymph nodes from 38 patients at Tenon Hospital were imaged minutes after excision with full-field optical coherence tomography in the pathology laboratory, before being handled for histological analysis. A pathologist performed a blind diagnosis (benign/malignant), based on the full-field optical coherence tomography images alone, which resulted in a sensitivity of 92% and a specificity of 83% (n = 65 samples). Regular feedback was given during the blind diagnosis, with thorough analysis of the images, such that features of normal and suspect nodes were identified in the images and compared with histology. A nonmedically trained imaging expert also performed a blind diagnosis aided by the reading criteria defined by the pathologist, which resulted in 85% sensitivity and 90% specificity (n = 71 samples). The number of false positives of the pathologist was reduced by 3 in a second blind reading a few months later. These results indicate that following adequate training, full-field optical coherence tomography can be an effective noninvasive diagnostic tool for extemporaneous sentinel node biopsy qualification.

Real-time cancer diagnosis during prostate biopsy: ex vivo evaluation of full-field optical coherence tomography (FFOCT) imaging on biopsy cores.

OBJECTIVES: To evaluate the diagnostic accuracy (Acc) of full-field optical coherence tomography (FFOCT) for cancer detection on prostate biopsy. MATERIALS AND METHODS: Thirty-eight consecutive patients with elevated PSA and/or suspicious digital rectal examination were prospectively included. For each patient, 1-10 cores were randomly selected and imaged with FFOCT immediately after sampling. The images obtained were de-identified and analyzed by three pathologists blinded to the results of pathological evaluation. The overall average Acc was measured, as well as sensitivity (Se), specificity (Sp), positive and negative predictive values (PPV and NPV). The Acc learning curve was assessed by multivariate logistic regression, and inter-reader concordance was assessed by Kappa index. RESULTS: One hundred and nineteen cores were imaged. Of them, 40 (33.6%) were involved with cancer. The overall average Acc of FFOCT for cancer detection was of 70.6%. Se, Sp, PPV, and NPV were of 63, 74, 55.5, and 80%, respectively. A substantial agreement was observed among pathologists (κ = 0.6, p < 0.001). On multivariate analysis, Acc was associated with the number of previously interpreted cases, with a predicted Acc of 82% at the end of learning curve. The overall average accuracy for high Gleason score (>3 + 3) determination was of 72%, although results were limited by the small amount of cases. CONCLUSIONS: FFOCT of prostate biopsy cores may provide a diagnostic accuracy greater than 80%, with a good reliability and a high NPV. TAKE HOME MESSAGE: "Full-field optical coherence tomography is a novel imaging modality that could have a potential value in real-time diagnosis of prostate cancer during prostate biopsy procedures."

A feasibility study of full-field optical coherence tomography for rapid evaluation of EUS-guided microbiopsy specimens.

BACKGROUND: Rapid on-site evaluation of cytologic specimens is a way of determining the adequacy of fine-needle aspiration (FNA). However, alternatives may be useful when the presence of a cytotechnologist and/or pathologist is not possible. OBJECTIVE: To evaluate the feasibility of using full-field optical coherence tomography (FFOCT) for FNA specimen quality assessment. DESIGN: FFOCT images were acquired on gastric, pancreatic, pelvic, and lymph-node formalin-fixed FNA specimens and were compared with histology of the same samples. SETTING: Pathology suite in a hospital. PATIENTS: Fourteen patients undergoing gastric, pancreatic, pelvic, or lymph-node EUS-guided FNA biopsy. INTERVENTIONS: FFOCT imaging on formalin-fixed samples before histologic procedures. MAIN OUTCOME MEASUREMENTS: FFOCT imaging feasibility and visibility of normal and abnormal features on images. RESULTS: FFOCT imaging was possible. Blood, mucus, muscle, collagen, and digestive mucosa could be identified as well as abnormal architectural features including infiltrative pancreatic ductal carcinoma and a neuroendocrine neoplasm. Lesions at the individual cell level could not be detected. LIMITATIONS: The study was performed on a limited number of cases. CONCLUSION: FFOCT offers rapid, noninvasive, nondestructive imaging of FNA biopsy specimens. In the future, it could be performed in the endoscopy suite to improve detection of satisfactory specimens and obviate the need for rapid on-site evaluation.

Large field, high resolution full-field optical coherence tomography: a pre-clinical study of human breast tissue and cancer assessment.

We present a benchmark pilot study in which high-resolution Full-Field Optical Coherence Tomography (FF-OCT) was used to image human breast tissue and is evaluated to assess its ability to aid the pathologist's management of intra-operative diagnoses. FF-OCT imaging safety was investigated and agreement between FF-OCT and routinely prepared histopathological images was evaluated. The compact setup used for this study provides 1 mm3 resolution and 200 mm imaging depth, and a 2.25 cm2 specimen is scanned in about 7 minutes. 75 breast specimens were imaged from 22 patients (21 women, 1 man) with a mean age of 58 (range: 25-83). Pathologists blind diagnosed normal/benign or malignant tissue based on FF-OCT images alone, diagnosis from histopathology followed for comparison. The contrast in the FF-OCT images is generated by intrinsic tissue scattering properties, meaning that no tissue staining or preparation is required. Major architectural features and tissue structures of benign breast tissue, including adipocytes, fibrous stroma, lobules and ducts were characterized. Subsequently, features resulting from pathological modification were characterized and a diagnosis decision tree was developed. Using FF-OCT images, two breast pathologists were able to distinguish normal/benign tissue from lesional with a sensitivity of 94% and 90%, and specificity of 75% and 79% respectively.

Full-field optical coherence tomography: a new technology for 3D high-resolution skin imaging.

BACKGROUND/AIMS: Full-field optical coherence tomography (FFOCT) is a new imaging technology that can provide 3D micron-level resolution and is suited for high-resolution imaging of biological tissue. The aim of this study was to evaluate its capacity and potential for imaging human epidermis and dermis and various skin pathologies in ex vivo and in vivo conditions. METHODS: Non-fixed and fixed samples of normal and pathological skin and normal in vivo skin were imaged with a FFOCT system and compared to histological slides. RESULTS: The epidermis and adnexae, the collagen bundles of the dermis and the hypodermis could be identified through architectural and cellular details. The pathological structures were distinguished from the normal structures and corresponded to their histopathological organization. CONCLUSION: FFOCT is a novel technology in the field of skin imaging that has the potential to be a relevant complement to existing non-invasive imaging modalities for clinical and cosmetic applications.

Role of ocular aberrations in photopic spatial summation in the fovea.

An adaptive optics vision simulator was used to measure the role of ocular aberrations in the photopic foveal spatial summation curve for three young subjects. After the correction of the ocular aberrations over a 6 mm pupil, the increment luminance threshold was lowered for small stimuli and the estimated area of complete summation (Ricco's area) was reduced by a factor of 2.6 on average. These results show that the image spread due to natural ocular aberrations partly accounts for the spatial summation observed in the fovea in the photopic light regime.

Experimental validation of a Bayesian model of visual acuity.

Based on standard procedures used in optometry clinics, we compare measurements of visual acuity for 10 subjects (11 eyes tested) in the presence of natural ocular aberrations and different degrees of induced defocus, with the predictions given by a Bayesian model customized with aberrometric data of the eye. The absolute predictions of the model, without any adjustment, show good agreement with the experimental data, in terms of correlation and absolute error. The efficiency of the model is discussed in comparison with image quality metrics and other customized visual process models. An analysis of the importance and customization of each stage of the model is also given; it stresses the potential high predictive power from precise modeling of ocular and neural transfer functions.

Correction of ocular and atmospheric wavefronts: a comparison of the performance of various deformable mirrors.

The main applications of adaptive optics are the correction of the effects of atmospheric turbulence on ground-based telescopes and the correction of ocular aberrations in retinal imaging and visual simulation. The requirements for the wavefront corrector, usually a deformable mirror, will depend on the statistics of the aberrations to be corrected; here we compare the spatial statistics of wavefront aberrations expected in these two applications. We also use measured influence functions and numerical simulations to compare the performance of eight commercially available deformable mirrors for these tasks. The performance is studied as a function of the size of the optical pupil relative to the actuated area of the mirrors and as a function of the number of modes corrected. In the ocular case it is found that, with the exception of segmented mirrors, the performance is greatly enhanced by having a ring of actuators outside the optical pupil, as this improves the correction of the pupil edge. The effect is much smaller in the case of Kolmogorov wavefronts. It is also found that a high Strehl ratio can be obtained in the ocular case with a relatively low number of actuators if the stroke is sufficient. Increasing the number of actuators has more importance in the Kolmogorov case, even for the relatively weak turbulence considered here.

Measured double-pass intensity point-spread function after adaptive optics correction of ocular aberrations.

The double-pass intensity point-spread function was recorded in four subjects using a monochromatic source emitting at 543 nm, through a 6.7-mm diameter pupil i) at the fovea after adaptive optics correction of the ocular aberrations, ii) at the fovea without adaptive optics correction, and iii) at 2 degrees of eccentricity with adaptive optics correction. The half-width at half-maximum of the double-pass point-spread function was narrower after correction of the ocular aberrations. At 2 degrees of eccentricity this width was larger than at the fovea. The minimum widths were about 1.1 arcmin in dark pigmented eyes and 1.6 arcmin in light pigmented eyes. These values are 6 to 9 times larger than the width expected from diffraction alone.

Use of a customized vision model to analyze the effects of higher-order ocular aberrations and neural filtering on contrast threshold performance.

Customized optical filtering and light-dependent neural filtering were implemented in an ideal-observer model for an L-alternative forced-choice visual task. The model was applied to a contrast threshold visual task with adaptive optics correction of ocular higher-order (HO) aberrations under different light regimes, for which experimental data have previously been obtained (J. Mod. Opt.55, 791, 2008). A separability measure was used to assess the model-observer performance and to investigate the joint effect of optical and neural filtering. The numerical results were consistent with the experimental data in the assessment of the effect of HO aberrations as a function of light level.

Comparative analysis of deformable mirrors for ocular adaptive optics.

We have evaluated the ability of three commercially available deformable mirrors to compensate the aberrations of the eye using a model for aberrations developed by Thibos, Bradley and Hong. The mirrors evaluated were a 37 actuator membrane mirror and 19 actuator piezo mirror (OKO Technologies) and a 35 actuator bimorph mirror (AOptix Inc). For each mirror, Zernike polynomials and typical ocular aberrated wavefronts were fitted with the mirror modes measured using a Twyman-Green interferometer. The bimorph mirror showed the lowest root mean square error, although the 19 actuator piezo device showed promise if extended to more actuators. The methodology can be used to evaluate new deformable mirrors as they become available.