Full-field Optical Coherence Tomography Research Articles

基于FPGA的快速全场光学相干层析方法

基于FPGA的快速全场光学相干层析方法

Rapid and high-resolution imaging of human liver specimens by full-field optical coherence tomography.

We report rapid and high-resolution tomographic en face imaging of human liver specimens by full-field optical coherence tomography (FF-OCT). First, the arrangement of the FF-OCT system was described and the performance of the system was measured. The measured axial and lateral resolutions of the system are 0.8 and 0.9 μm, respectively. The system has a sensitivity of ∼60 dB and can achieve an imaging rate of 7 fps and a penetration depth of ∼80 μm. The histological structures of normal liver can be seen clearly in the en face tomographic images, including central veins, cords of hepatocytes separated by sinusoidal spaces, and portal area (portal vein, the hepatic arteriole, and the bile duct). A wide variety of histological subtypes of hepatocellular carcinoma was observed in en face tomographic images, revealing notable cancerous features, including the nuclear atypia (enlarged convoluted nuclei), the polygonal tumor cells with obvious resemblance to hepatocytes with enlarged nuclei. In addition, thicker fibrous bands, which make the cytoplasmic plump vesicular nuclei indistinct, were also seen in the images. Finally, comparison between the portal vein in a normal specimen versus that seen in the rare type of cholangiocarcinoma was made. The results show that the cholangiocarcinoma presents with a blurred pattern of portal vein in the lateral direction and an aggregated distribution in the axial direction; the surrounding sinusoidal spaces and nuclei of cholangiocarcinoma are absent. The findings in this work may be used as additional signs of liver cancer or cholangiocarcinoma, demonstrating capacity of FFOCT device for early cancer diagnosis and many other tumor-related studies in biopsy.

Image contrast reduction mechanism in full-field optical coherence tomography.

Correct interpretation of image contrast obtained with full-field optical coherence tomography (FFOCT) technique is required for accurate medical diagnosis applications. In this work, first, the characteristics of microscopic structures of tissue that generate the contrast in en-face tomographic image obtained with FFOCT are discussed. Then an overview is given of the parameters that affect image contrast. Finally, the contrast correction factor for correct image interpretation and the contrast limits to practical FFOCT systems are outlined.

Fingerprint imaging from the inside of a finger with full-field optical coherence tomography.

Imaging below fingertip surface might be a useful alternative to the traditional fingerprint sensing since the internal finger features are more reliable than the external ones. One of the most promising subsurface imaging technique is optical coherence tomography (OCT), which, however, has to acquire 3-D data even when a single en face image is required. This makes OCT inherently slow for en face imaging and produce unnecessary large data sets. Here we demonstrate that full-field optical coherence tomography (FF-OCT) can be used to produce en face images of sweat pores and internal fingerprints, which can be used for the identification purposes.

Control of 3D IC process steps by optical metrologies

This paper evaluates various optical metrology techniques for in-line control of the uniformity of 3D stacked structures. Advanced packaging technologies are rapidly evolving and 3D architectures require very well controlled process steps. Optical metrology techniques are now used for TSV but also for interconnect processing. In engineering mode, at the process implementation step, these techniques must be evaluated and then used to get uniform and repeatable processes. Among the 3D TSV Via middle process flow, the temporary bonding, wafer thinning and TSV reveal are key 3D process steps to get a uniform backside copper nail signature. High aspect ratio TSV are etched in Front End of the line and deep reactive ion etching systems generates radial non uniformities signatures. The wafer backside processing challenge consists in compensating this issue among the entire wafer surface. In this paper, all these process steps were characterized in order to quantify their specific intra-wafer dispersion signature .of the related key parameters. Cross correlation between the various intra-wafer process step signatures was then analyzed to verify the data set consistency and several process parameters analyzed to get a simple model. This model was then used to get a very uniform copper nail signature. For small diameter TSV and corresponding copper nails, a comparison has been done between full field OCT and confocal chromatic techniques.

Label-free characterization of vitrification-induced morphology changes in single-cell embryos with full-field optical coherence tomography.

Vitrification is an increasingly popular method of embryo cryopreservation that is used in assisted reproductive technology. Although vitrification has high post-thaw survival rates compared to other freezing techniques, its long-term effects on embryo development are still poorly understood. We demonstrate an application of full-field optical coherence tomography (FF-OCT) to visualize the effects of vitrification on live single-cell (2 pronuclear) mouse embryos without harmful labels. Using FF-OCT, we observed that vitrification causes a significant increase in the aggregation of structures within the embryo cytoplasm, consistent with reports in literature based on fluorescence techniques. We quantify the degree of aggregation with an objective metric, the cytoplasmic aggregation (CA) score, and observe a high degree of correlation between the CA scores of FF-OCT images of embryos and of fluorescence images of their mitochondria. Our results indicate that FF-OCT shows promise as a label-free assessment of the effects of vitrification on embryo mitochondria distribution. The CA score provides a quantitative metric to describe the degree to which embryos have been affected by vitrification and could aid clinicians in selecting embryos for transfer.

Full-Field Swept Source Master-Slave Optical Coherence Tomography

We apply the principle of master-slave (MS) interferometry to a full-field swept source optical coherence tomography (OCT) setup equipped with a fast 2-D camera. MS interferometry does not involve Fourier transformations and, therefore, eliminates the need for spectrum data resampling required by the conventional spectral domain OCT. Using this method in a full-field OCT setup, en face images are acquired in vivo from finger skin, Drosophila melanogaster larvae, and pupae, with no spectrum resampling and no mechanical scanning. The signal processing is based on a comparison operation of the shapes of channeled spectra for each camera pixel, with a set of reference signals (masks). In this way, en face OCT images can be obtained with no need for the volumetric assembly and software cutting the en face images from an image volume, which are required by the conventional spectral domain OCT method. We demonstrate that the sensitivity and axial resolution of the MS method in a full-field swept source OCT setup are similar to the values obtained using the conventional Fourier-transformation-based swept source OCT method in a full-field setup. Multiple en face images can be produced in parallel by using multiple stored shapes of channeled spectra for the depths of interest. The full-field MS-OCT method presented here opens the possibility of parallel processing for all image points in a 3-D volume of the object.

Dark-field full-field optical coherence tomography.

Full-field optical coherence tomography (FF-OCT) provides en face images from deep in the tissue with high spatial resolution. Specular reflections, however, may reduce image contrast as it can be much stronger than the backscattered signal from a specimen. To this end, we demonstrate dark-field FF-OCT (d-FF-OCT) that can block specular reflections by the help of an opaque disk in the pupil-conjugated plane. The reference mirror is replaced by a blazed grating, which eliminates a walk-off between the sample and the reference beams on a camera that otherwise limits the imaging field-of-view (FOV). We show that d-FF-OCT can suppress specular reflections efficiently from the glass-specimen interface by at least two orders of magnitude and yield higher contrast images compared to the conventional FF-OCT.

Fourier spectrum analysis of full-field optical coherence tomography for tissue imaging

We propose a model of the full-field optical coherence tomography (FFOCT) technique for tissue imaging, in which the fractal model of the spatial correlation function of the refractive index of tissue is employed to approximate tissue structure. The results may be helpful for correctly interpreting en face tomographic images obtained with FFOCT.

Elastography of multicellular aggregates submitted to osmo-mechanical stress

Tumors are subjected to mechanical stress generated by their own growth in a confined environment, and by their surrounding tissues. Recent works have focused on the study of the growth of spherical aggregates of cells, spheroids, under controlled confinement or stress. In this study we demonstrate the measurement of spatially and temporally resolved deformation maps inside spheroids while applying an osmo-mechanical stress. We use full field optical coherence tomography, a high resolution imaging technique well suited for real-time measurements of deformation in living tissues under stress. Using the spherical symmetry of the experiment, we compare our data to a mechanical modeling of the spheroid as a continuous medium. We estimate the viscoelastic parameters of spheroids and discuss the apparent tissue anisotropy after the osmo-mechanical stress.

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

To evaluate the diagnostic accuracy (Acc) of full-field optical coherence tomography (FFOCT) for cancer detection on prostate biopsy. 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. 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. FFOCT of prostate biopsy cores may provide a diagnostic accuracy greater than 80%, with a good reliability and a high NPV. "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."

Applying LED in full-field optical coherence tomography for gastrointestinal endoscopy

Optical coherence tomography (OCT) has become an important medical imaging technology due to its non-invasiveness and high resolution. Full-field optical coherence tomography (FF-OCT) is a scanning scheme especially suitable for en face imaging as it employs a CMOS/CCD device for parallel pixels processing. FF-OCT can also be applied to high-speed endoscopic imaging. Applying cylindrical scanning and a right-angle prism, we successfully obtained a 360° tomography of the inner wall of an intestinal cavity through an FF-OCT system with an LED source. The 10-μm scale resolution enables the early detection of gastrointestinal lesions, which can increase detection rates for esophageal, stomach, or vaginal cancer. All devices used in this system can be integrated by MOEMS technology to contribute to the studies of gastrointestinal medicine and advanced endoscopy technology.

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.

Rapid evaluation of fresh ex vivo kidney tissue with full-field optical coherence tomography

Background: Full-field optical coherence tomography (FFOCT) is a real-time imaging technique that rapidly generates images reminiscent of histology without any tissue processing, warranting its exploration for evaluation of ex vivo kidney tissue. Methods: Fresh tissue sections from tumor and adjacent nonneoplastic kidney (n = 25 nephrectomy specimens; clear cell renal cell carcinoma (CCRCC) = 12, papillary RCC (PRCC) = 4, chromophobe RCC (ChRCC) = 4, papillary urothelial carcinoma (PUC) = 1, angiomyolipoma (AML) = 2 and cystic nephroma = 2) were imaged with a commercial FFOCT device. Sections were submitted for routine histopathological diagnosis. Results: Glomeruli, tubules, interstitium, and blood vessels were identified in nonneoplastic tissue. In tumor sections, the normal architecture was completely replaced by either sheets of cells/trabeculae or papillary structures. The former pattern was seen predominantly in CCRCC/ChRCC and the latter in PRCC/PUC (as confirmed on H&E). Although the cellular details were not very prominent at this resolution, we could identify unique cytoplasmic signatures in some kidney tumors. For example, the hyper-intense punctate signal in the cytoplasm of CRCC represents glycogen/lipid, large cells with abundant hyper-intense cytoplasm representing histiocytes in PRCC, and signal-void large polygonal cell representing adipocytes in AML. According to a blinded analysis was performed by an uropathologist, all nonneoplastic tissues were differentiated from neoplastic tissues. Further, all benign tumors were called benign and malignant were called malignant. A diagnostic accuracy of 80% was obtained in subtyping the tumors. Conclusion: The ability of FFOCT to reliably differentiate nonneoplastic from neoplastic tissue and identify some tumor types makes it a valuable tool for rapid evaluation of ex vivo kidney tissue e.g. for intraoperative margin assessment and kidney biopsy adequacy. Recently, higher resolution images were achieved using an experimental FFOCT setup. This setup has the potential to further increase the diagnostic accuracy of FFOCT.

提高全场光学相干层析系统成像质量的方法

提高全场光学相干层析系统成像质量的方法

Anisotropic polyvinyl alcohol hydrogel phantom for shear wave elastography in fibrous biological soft tissue: a multimodality characterization

Shear wave elastography imaging techniques provide quantitative measurement of soft tissues elastic properties. Tendons, muscles and cerebral tissues are composed of fibers, which induce a strong anisotropic effect on the mechanical behavior. Currently, these tissues cannot be accurately represented by existing elastography phantoms. Recently, a novel approach for orthotropic hydrogel mimicking soft tissues has been developed (Millon et al J. Biomed. Mater. Res. B 305–11). The mechanical anisotropy is induced in a polyvinyl alcohol (PVA) cryogel by stretching the physical crosslinks of the polymeric chains while undergoing freeze/thaw cycles. In the present study we propose an original multimodality imaging characterization of this new transverse isotropic (TI) PVA hydrogel. Multiple properties were investigated using a large variety of techniques at different scales compared with an isotropic PVA hydrogel undergoing similar imaging and rheology protocols. The anisotropic mechanical (dynamic and static) properties were studied using supersonic shear wave imaging technique, full-field optical coherence tomography (FFOCT) strain imaging and classical linear rheometry using dynamic mechanical analysis. The anisotropic optical and ultrasonic spatial coherence properties were measured by FFOCT volumetric imaging and backscatter tensor imaging, respectively. Correlation of mechanical and optical properties demonstrates the complementarity of these techniques for the study of anisotropy on a multi-scale range as well as the potential of this TI phantom as fibrous tissue-mimicking phantom for shear wave elastographic applications.

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

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. To evaluate the feasibility of using full-field optical coherence tomography (FFOCT) for FNA specimen quality assessment. FFOCT images were acquired on gastric, pancreatic, pelvic, and lymph-node formalin-fixed FNA specimens and were compared with histology of the same samples. Pathology suite in a hospital. Fourteen patients undergoing gastric, pancreatic, pelvic, or lymph-node EUS-guided FNA biopsy. FFOCT imaging on formalin-fixed samples before histologic procedures. FFOCT imaging feasibility and visibility of normal and abnormal features on images. 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. The study was performed on a limited number of cases. 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.

Full-Field Optical Coherence Tomography of Human Donor and Pathological Corneas

Purpose: To evaluate the performance of a full-field optical coherence tomography (FF-OCT) system in the study of human donor and pathological corneas and assess its suitability for use in eye banks.Methods: Our study was carried out using an FF-OCT system developed for non-invasive imaging of tissue structures in depth with ultrahigh resolution (1 µm in all directions). Images were acquired from eight stored human donor corneas (either edematous or after deswelling) and five surgical specimens of corneas with various diseases (bullous keratopathy, lattice corneal dystrophy, stromal scar after keratitis, keratoconus and Fuchs dystrophy). They were compared with standard histology and pre-operative spectral domain OCT.Results: The FF-OCT device enabled a precise visualization of the cells and the different structures (epithelium, basement membrane, Bowman’s layer, stroma, Descemet’s membrane and endothelium) in normal corneas. Specific lesions in various corneal diseases could also be easily identified, such as corneal edema, epithelium and Bowman’s layer irregularities, breaks, or scars (keratoconus), stromal opacities, deposits, fibrosis (stromal corneal scar, bullous keratopathy, lattice corneal dystrophy) and Descemet’s membrane thickening and guttae (Fuchs dystrophy). FF-OCT image features were comparable to the details provided by conventional histology. Higher resolution could be demonstrated with FF-OCT when compared with spectral domain OCT.Conclusion: FF-OCT is a powerful non-invasive imaging tool that allows detailed study of corneal structures. Images correlate well with conventional histology. Further studies should evaluate the benefit of this technique as a complement to current assessment methods of human donor corneas.

Detection of plasmonic nanoparticles with full field-OCT: optical and photothermal detection.

Detecting the signal backscattered by nanoparticles immersed in highly scattering media such as biological tissue remains a challenge. In this article we report on the use of Full Field OCT (FF-OCT) to slice in depth in phantoms and in tissues in order a) to selectively observe the particles through the backscattered light at suitable wavelengths, and b) to detect the effects of the time-dependent response to full field optical heating through the strong absorption cross-section of these plasmonic nanoparticles. The analysis of the thermal wave behavior leads to the localization of the heat sources even when FF-OCT signals cannot reach the heated area.

Effects of temporal and spatial coherence on resolution in full-field optical coherence tomography

The contrast enhanced imaging of microscopic structures of tissue and the high-resolution optical sectional imaging of thin slices within tissue are two main advantages of full-field optical coherence tomography (FFOCT) over the conventional optical microscopy. In this work, first a detailed description of the development of FFOCT technique is reviewed. The imaging signal model of FFOCT is then described, which reveals the roles of both the light-intensity distribution in the pupil plane of the objective lenses and the light spectrum of the source on the system resolution. Then the effects of the temporal and spatial coherence on the lateral resolution and depth resolution of FFOCT are considered. Finally, the change of separation between the temporal coherence envelope and the longitudinal spatial coherence envelope is analyzed induced by the mismatch of the refractive index of tissue and the surrounding medium.