LC-OCT is a recently developed technology for the diagnosis and monitoring of skin pathologies. 3D LC-OCT images with isotropic cellular resolution allow visualization of skin layers and tissue characterization at the cellular level. A dermoscopic imaging system has been coupled to LC-OCT, allowing localization and coverage of lesions, together with characterization of their margins. Finally, segmentations of the epidermal layers and keratinocyte nuclei performed using artificial intelligence allow diagnosis and follow-up of the treatment of skin pathologies in a non-invasive and quantitative way. These tools could help improve the accuracy of clinical diagnosis, allowing early detection of malignant skin tumors.
Line-field Confocal Optical Coherence Tomography (LC-OCT) is a non-invasive optical technique for imaging the skin in vivo in depth and at high resolution. Confocal Raman Microspectroscopy (CRM) is a label-free technique that provides point-wise information on the molecular content of the analyzed sample. We present the development of a prototype platform allowing the co-localized acquisition of LC-OCT images and Raman spectra on ex vivo skin samples with an accuracy of ± 20 μm. We present results obtained on healthy skin as well as preliminary results obtained on a basal cell carcinoma surgery, aiming at identifying molecular tumor markers.
KEYWORDS: Skin, Optical coherence tomography, Confocal microscopy, In vivo imaging, Imaging systems, 3D image processing, Target acquisition, Reflectivity, Real time imaging, Optical properties
A combined LC-OCT and dermoscopy system is presented. The LC-OCT modality allows to obtain real-time in-depth cross-sectional and 3D images with an isotropic cellular resolution, providing a comprehensive view of the in-depth tridimensional morphology of the skin at a resolution similar to histology. The dermoscopy modality provides color images of the surface of the skin, serving as a guiding tool for LC-OCT, and complementing the in-depth cellular information of LC-OCT with a high-resolution view of the skin surface. The system has been applied to in vivo imaging of various skin lesions, including carcinomas and melanomas.
Significance: Line-field confocal optical coherence tomography (LC-OCT) is a recently introduced high-resolution imaging modality based on a combination of low-coherence optical interferometry and reflectance confocal optical microscopy with line illumination and line detection. Capable of producing three-dimensional (3D) images of the skin with cellular resolution, in vivo, LC-OCT has been mainly applied in dermatology and dermo-cosmetology. The LC-OCT devices capable of acquiring 3D images reported so far are based on a Linnik interferometer using two identical microscope objectives. In this configuration, LC-OCT cannot be designed to be a very compact and light device, and the image acquisition speed is limited.Aim: The objective of this work was to develop a more compact and lighter LC-OCT device that is capable of acquiring images faster without significant degradation of the resolution and with optimized detection sensitivity.Approach: We developed an LC-OCT device based on a Mirau interferometer using a single objective. Dynamic adjustment of the camera frequency during the depth scan is implemented, using a faster camera and a more powerful light source. The reflectivity of the beam-splitter in the Mirau interferometer was optimized to maximize the detection sensitivity. A galvanometer scanner was incorporated into the device for scanning the illumination line laterally. A stack of adjacent B-scans, constituting a 3D image, can thus be acquired.Results: The device is able to acquire and display B-scans at 17 fps. 3D images with a quasi-isotropic resolution of ∼1.5 μm (1.3, 1.9, and 1.1 μm in the x , y, and z directions, respectively) over a field of 940 μm × 600 μm × 350 μm (x × y × z) can be obtained. 3D imaging of human skin at cellular resolution, in vivo, is reported.Conclusions: The acquisition rate of the B-scans, at 17 fps, is unprecedented in LC-OCT. Compared with the conventional LC-OCT devices based on a Linnik interferometer, the reported Mirau-based LC-OCT device can acquire B-scans ∼2 times faster. With potential advantages in terms of compactness and weight, a Mirau-based device could easily be integrated into a smaller and lighter handheld probe for use by dermatologists in their daily medical practice.
Line-field confocal optical coherence tomography (LC-OCT) is a high-resolution imaging technique based on a combination of time-domain optical coherence tomography and confocal optical microscopy, with line illumination using a spatially coherent broadband light source and line detection using a line camera. We present a LC-OCT device based on a Mirau interferometer consisting of an immersion microscope objective incorporating a miniature interferometer. The device can acquire 17 B-scans per second, which is the fastest acquisition rate reported to date in LC-OCT. By stacking multiple adjacent B-scans, a 3D image with a lateral field of view of 940 μm × 600 μm over a depth of 350 μm can be acquired. Compared to the conventional LC-OCT devices based on a Linnik interferometer, this Mirau-based device has advantages in terms of compactness, weight, and B-scan acquisition speed. Imaging of skin tissue with near-isotropic resolution of ~1.5 micron is demonstrated in vivo.
Line-field confocal optical coherence tomography (LC-OCT) is an imaging technique based on a combination of confocal microscopy and OCT, allowing three-dimensional cellular-resolution imaging of the skin in vivo. We present the latest advances in LC-OCT to facilitate the use of the technique by dermatologists and improve the diagnosis and analysis of skin lesions. A video camera was incorporated into a handheld probe to acquire dermoscopic images in parallel with LCOCT images. A confocal Raman spectrometer was associated with a LC-OCT device to record morphological images of the skin in which points of interest can be subjected to molecular characterization.
Line-field confocal optical coherence tomography (LC-OCT) is an optical technique generating three-dimensional (3D) images of the skin at cellular resolution. Confocal Raman microspectroscopy (CRM) is an optical modality that provides a point-wise molecular fingerprint of samples. We have developed a method to co-localize data acquired by separate LC-OCT and CRM systems. LC-OCT allows for recording 3D morphological overview images in which points of interest (POIs) can be localized for molecular analysis using CRM. A biopsy of skin with a red-colored tattoo with unknown ink composition was analyzed using co-localized LC-OCT and CRM. After acquisition of a 3D LC-OCT image, specific POIs were targeted inside the biopsy, based on their morphological features: POIs located in the epidermis where no ink was expected to be found as well as POIs located in bright areas of the dermis, down to 275 μm in depth, likely indicating the presence of tattoo ink. Analysis of the spectra at these specific POIs confirmed the absence of tattoo ink in the epidermis and its presence in the bright areas in the dermis. A stronger molecular signal of ink in the brightest areas of the dermis identified with LC-OCT was also revealed, suggesting a higher amount of ink. The combination and co-localization of LC-OCT and CRM brings a new level of characterization of the skin, enabling molecular analysis of POIs based on their morphological aspect at cellular-resolution within a volume, which could be of great interest in dermatology and dermo-cosmetics.
KEYWORDS: Skin, Image segmentation, 3D modeling, 3D image processing, In vivo imaging, Optical coherence tomography, Artificial intelligence, Confocal microscopy, 3D acquisition
Line-field confocal optical coherence tomography (LC-OCT) is an imaging technique based on a combination of reflectance confocal microscopy and optical coherence tomography, allowing three-dimensional (3D) imaging of skin in vivo with an isotropic spatial resolution of about 1.3 micron and up to 400 microns in depth. Cellular-resolution 3D images obtained with LC-OCT offer a considerable amount of information for description and quantification of the upper layers of in vivo skin using morphological metrics, which can be critical for better understanding the skin changes leading to aging or some pathologies. This study introduces metrics for the quantification of the epidermis, and uses them to describe the variability of healthy epidermis between different body sites. These metrics include the stratum corneum thickness, the undulation of the dermal-epidermal junction (DEJ), and the quantification of the keratinocyte network. In order to generate relevant metrics over entire 3D images, an artificial intelligence approach was applied to automate the calculation of the metrics. We were able to quantify the epidermis of eight volunteers on seven body areas on the head, the upper limbs and the trunk. Epidermal thicknesses and DEJ undulation variations were observed between different body sites. The cheek presented the thinnest stratum corneum the least undulated DEJ, while the back of the hand presented the thickest stratum corneum and the back the most undulated DEJ. The process of keratinocyte maturation was evidenced in vivo. These 3D in vivo quantifications open the door in clinical practice to diagnose and monitor pathologies for which the epidermis is impaired.
Line-field confocal optical coherence tomography (LC-OCT) is an optical technique based on a combination of reflectance confocal microscopy and optical coherence tomography to generate cellular-resolution images, in either vertical (x×z) or horizontal (x×y) sections, or in three-dimensions (3D), with a field of view of 1.2×0.5×0.4 mm3 (x×y×z). LC-OCT was originally designed for in vivo skin imaging. Here, we present a novel implementation of LC-OCT that enables ex vivo biological tissue imaging with an extended field of view. In this implementation, a specific sample holder is used so that the head of the LC-OCT device is not in contact with the sample to be imaged. The sample can thus be displaced independently of the LC-OCT head in a controlled manner using multi-axes motorized translations, while acquiring LCOCT images. A stitching algorithm is used to extend the field of view of vertical section images, horizontal section images, and also 3D images. In addition, as the device can also acquires color images of the sample surface (dermoscopic images) in parallel with the tomographic LC-OCT images, a similar mosaicking approach can be applied to the surface images. The method allows the reconstruction of a wide-field surface image of the sample, precisely collocated to the LC-OCT mosaic. This approach allows full characterization of entire skin punch biopsies of several millimeters with ~ 1 micron resolution, in 3D, and over a depth of 0.4 mm, with an associated dermoscopy-like image of the entire tissue surface.
Line-field confocal optical coherence tomography (LC-OCT) is an imaging technique that combines the principles of time-domain OCT and reflectance confocal microscopy (RCM). LC-OCT was designed to generate threedimensional (3D) morphological images of the skin, in vivo, with a spatial resolution of ∼ 1 μm. As in OCT and RCM, LC-OCT image contrast originates from the backscattering of incident light by the sample microstructures, which is determined by the optical scattering properties of the sample, namely the scattering coefficient μs and the scattering anisotropy parameter g. When imaging biological tissues, these properties can provide insight into tissue organization and structure, and could be used for quantitative tissue characterization in vivo. We present a method for obtaining spatially-resolved measurements of optical scattering parameters from LC-OCT images. Our approach is based on a calibration using a test sample with known optical scattering properties and on the application of a theoretical model previously developed for focus-tracking mode OCT and RCM. Assuming a single-scattering regime, this model allows to derive the optical scattering parameters μs and g from the intensity depth profiles acquired by LC-OCT. Spatially-resolved measurements are achieved by dividing the 3D LC-OCT image into “macro-voxels” and analyzing the different sample layers separately, leading to 3D distributions of μs and g. This method was experimentally tested against integrating spheres and collimated transmission measurements and validated on a set of mono- and bi-layered scattering phantoms.
Line-field confocal optical coherence tomography (LC-OCT) is an optical technology developed for in vivo skin imaging. Combining the principles of OCT and reflectance confocal microscopy, LC-OCT generates high-resolution three-dimensional (3D) images with an isotropic spatial resolution of 1.3 micron and up to 500 microns in depth. Confocal Raman microspectroscopy is a label-free optical technique which allows for point measurement of the molecular content of a sample with micrometer resolution. While LC-OCT provides morphological information, Raman spectroscopy brings chemical information but lacks image guidance for targeting specific points of interest in the sample. Combining the two modalities would therefore provide complementary information and guidance for Raman measurements. We present a method to co-localize LC-OCT and Raman acquisitions for ex vivo applications. This co-localization approach allows acquisition of Raman spectra at specific locations targeted in a 3D LC-OCT image, with an accuracy of ± 20 μm. The co-localization method was developed using a LC-OCT device designed for ex vivo imaging and a custom Raman system. The principle of co-localization relies on both the use of a specific sample holder that can be positioned under each device with high repeatability and of a coordinate-based calibration between the two devices. Co-localization was validated using pig skin samples containing tattoo ink of known composition. LC-OCT images allowed to target specific regions in the samples where the presence of tattoo ink was revealed by detection of a specific Raman signature.
This paper reports on line-field confocal optical coherence tomography (LC-OCT), a recently invented imaging technology now capable of generating either a horizontal (en face) section image at an adjustable depth, or a vertical section image (B-scan) at an adjustable lateral position. For both operating modes, images are acquired in real-time (10 frames/second), with real-time control of the depth and lateral positions. Using a supercontinuum laser as a broadband light source and a high numerical microscope objective, an isotropic spatial resolution of ∼ 1 μm is achieved. The imaging fields of view are 1.2×0.5 mm² (x×y, horizontal) and 1.2×0.5 mm² (x×z, vertical). LC-OCT has been used in dermatology for skin imaging.
Line-field confocal optical coherence tomography (LC-OCT) is an imaging technique based on time-domain OCT with line illumination and line detection. The focus is adjusted during the scan of the sample depth to image with high lateral resolution (~ 1 μm), similar to the axial resolution, at a central wavelength of ~ 800 nm. The LC-OCT prototypes reported so far were all based on a Linnik-type interferometer. We present in this paper a LC-OCT device based on a Mirau interferometer. This Mirau-based LC-OCT device has the advantage of being more compact and lighter. In vivo imaging of human skin with a resolution of 1.3 μm × 1.1 μm (lateral × axial) is demonstrated at 12 frames per second over a field of 0.9 mm × 0.4 mm (lateral × axial).
Line-field confocal optical coherence tomography (LC-OCT) is an imaging method based on dynamically-focused line-field time-domain OCT to generate vertical section images of biological tissues with cellular spatial resolution. We demonstrate here an improvement in the LC-OCT technology that makes it possible to also generate horizontal (en face) section images, as in conventional optical coherence microscopy (OCM) or reflectance confocal microscopy (RCM). This improvement is based on the implementation of a lateral scanning device using a mirror galvanometer to scan the laser beam laterally at a given depth. The reference mirror of the interferometer is mounted on a piezoelectric transducer to generate a sinusoidal phase modulation during lateral scanning in order to extract the tomographic signal. The LC-OCT device is capable of generating either a horizontal section image at an adjustable depth in the sample, or a vertical section image at an adjustable lateral position in the sample. For both operating modes, images are acquired in real-time (8 frames per second), with real-time control of the depth/lateral position. The user can switch between the two operating modes by clicking on a button. Using a supercontinuum laser as a light source and 0.5 NA microscope objectives, the image resolution is 0.9 × 0.9 × 1.1 μm3 (x × y × z), with fields of view of 1.2 × 0.5 mm2 (x × y, horizontal) and 1.2 × 0.4 mm2 (x × z, vertical). In vivo cellular-level imaging of human skin is demonstrated for both modes of operation.
Line-field confocal optical coherence tomography (LC-OCT) is an imaging method based on dynamically-focused line-field time domain OCT to generate cellular resolution images of biological tissues, either in vertical sections (cross-sectional) or in horizontal sections (en face), with field of views of 1.2 × 0.5 mm2 (horizontal) and 1.2 × 0.4 mm2 (vertical). A handheld LC-OCT probe has recently been developed to facilitate the use of this imaging technique in dermatology for non invasive detection of skin cancers. We present here a video mosaicking method to reconstruct LC-OCT images with extended fields of view from a sequence of images obtained by a free displacement by the user of the LC-OCT probe on the skin surface. The lateral field of view is extended in both vertical and horizontal section images to ~ 5 mm, while maintaining an isotropic spatial resolution of ~ 1 μm. LC-OCT imaging with video-mosaicking is demonstrated for following the edges of large skin structure, as a proof-of-principle of in vivo tumor margin delineation, and for extending the lateral field of vertical section images, in order to approach the field of view of histology in this direction.
The use of non-invasive imaging techniques in dermatology has been reported to improve the diagnostic accuracy and the practice of biopsies, and at the same time to reduce the need for tissue excision. However, the current clinically-available imaging techniques do not yet entirely meet the need for early and accurate, non-invasive detection of all skin cancers. A handheld line-field confocal optical coherence tomography (LC-OCT) device has been designed for high-resolution non-invasive imaging of human skin, in vivo. LC-OCT delivers tomographic images in real-time (10 frames/s) with a quasi isotropic spatial resolution of ~ 1 μm, revealing a comprehensive morphological mapping of skin tissues at a cellular level, down to a depth of ~500 μm. The device has been applied to the in vivo imaging of various skin lesions. Surgical excisions of the lesions have then been performed followed by tissue processing to realize H&E-stained histopathological images. The spatial resolution, orientation, and imaging contrast mechanism of the LC-OCT images have allowed for a good level of similarity with the conventional histopathological images. LC-OCT was able to show most of the histopathological elements that allow for medical diagnosis. Using handheld LC-OCT as an adjunct tool in dermatology could help improve clinical diagnostic accuracy, allowing for the early detection of malignant skin tumors and a reduction in the number of surgical excisions of benign lesions.
We present an improved time-domain optical coherence tomography technique designed for ultrahigh-resolution B-scan imaging in real-time. The technique, called line-field confocal optical coherence tomography, is based on a Linnik-type interference microscope with line illumination using a supercontinuum laser and line detection using a line-scan camera. Bscan imaging with dynamic focusing is achieved by acquiring multiple A-scans in parallel. In vivo cellular level resolution imaging of skin is demonstrated at 10 frame/s with a penetration depth of ∼ 500 μm, with a spatial resolution of 1.3 μm × 1.1 μm (transverse × axial).
This work reports on the feasibility of dynamic imaging using conventional reflectivity-based tomographic images
obtained with full-field optical coherence microscopy (FF-OCM). Implementation of speckle variance for flow mapping
with an imaging rate of 180 Hz is demonstrated by mapping 20% intralipid flowing into 100-μm-diameter microcapillary
tubes at speeds up to ~ 50 mm/s. This constitutes a significant advance in high-resolution, real-time microvasculature
mapping, using FF-OCM. The acquisition scheme in FF-OCM is particularly appropriate for en face visualization of the
microvasculature, as FF-OCM directly acquires en face tomographic images unlike conventional OCT which usually
requires reslicing of a three-dimensional data set to get en face images.
This work reports on a compact full-field optical coherence microscopy (FF-OCM) setup specifically designed to meet
the needs for in vivo imaging, illuminated by a high-brightness broadband light emitting diode (LED). Broadband LEDs
have spectra potentially large enough to provide imaging spatial resolutions similar to those reached using conventional
halogen lamps, but their radiance can be much higher, which leads to high speed acquisition and makes in vivo imaging
possible. We introduce a FF-OCM setup using a 2.3 W broadband LED, with an interferometer designed to be as
compact as possible in order to provide the basis for a portable system that will make it possible to fully benefit from the
capacity for in vivo imaging by providing the ability to image any region of interest in real-time. The interferometer part
of the compact FF-OCM setup weighs 210 g for a size of 11x11x5 cm3. Using this setup, a sub-micron axial resolution
was reached, with a detection sensitivity of 68 dB at an imaging rate of 250 Hz. Due to the high imaging rate, the
sensitivity could be improved by accumulation while maintaining an acquisition time short enough for in vivo imaging. It
was possible to reach a sensitivity of 75 dB at a 50 Hz imaging rate. High resolution in vivo human skin images were
obtained with this setup and compared with images of excised human skin, showing high similarity.
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