Confocal Optical Coherence Tomography Research Articles

Digital imaging of ink spot lentigo: Confocal microscopy and line‐field confocal optical coherence tomography assessment

Digital imaging of ink spot lentigo: Confocal microscopy and line‐field confocal optical coherence tomography assessment

Corneal Nerve Assessment by Aesthesiometry: History, Advancements, and Future Directions.

The measurement of corneal sensation allows clinicians to assess the status of corneal innervation and serves as a crucial indicator of corneal disease and eye health. Many devices are available to assess corneal sensation, including the Cochet-Bonnet aesthesiometer, the Belmonte Aesthesiometer, the Swiss Liquid Jet Aesthesiometer, and the newly introduced Corneal Esthesiometer Brill. Increasing the clinical use of in vivo confocal microscopy and optical coherence tomography will allow for greater insight into the diagnosis, classification, and monitoring of ocular surface diseases such as neurotrophic keratopathy; however, formal esthesiometric measurement remains necessary to assess the functional status of corneal nerves. These aesthesiometers vary widely in their mode of corneal stimulus generation and their relative accessibility, precision, and ease of clinical use. The development of future devices to optimize these characteristics, as well as further comparative studies between device types should enable more accurate and precise diagnosis and treatment of corneal innervation deficits. The purpose of this narrative review is to describe the advancements in the use of aesthesiometers since their introduction to clinical practice, compare currently available devices for assessing corneal innervation and their relative limitations, and discuss how the assessment of corneal innervation is crucial to understanding and treating pathologies of the ocular surface.

Criteria for melanocytic lesions in LC-OCT.

Line-field confocal optical coherence tomography (LC-OCT) is an emerging diagnostic tool with imaging depth reaching ~400 μm and a novel three-dimensional (3D) cube providing cellular resolution. As far as we are aware, there are only a limited number of papers that have reported diagnostic criteria for melanocytic lesions using this technique, and none of them have been multicentric. Our aim was to establish the diagnostic criteria for melanocytic lesions using LC-OCT and identify the most significant architectural and cytologic features associated with malignancy. A retrospective evaluation of 80 consecutive melanocytic lesions from a prospective multicentric data set spanning three European centres was conducted. We excluded facial, acral and mucosal lesions from the study. Dermoscopic and LC-OCT images were evaluated by a consensus of four observers. Multivariate logistic regression with backward elimination was employed. The main melanoma diagnostic criteria include detecting >10 pagetoid cells in 3D acquisition, irregular 3D epidermal architecture, disrupted dermoepidermal junction (DEJ) and clefting. Significant risk factors were irregular 3D epidermal architecture, >10 pagetoid cells, dendritic cells at DEJ without underlying inflammation. Novel malignancy criteria in vertical view were DEJ disruption and clefting around atypical melanocyte nests. Exclusive melanoma features were epidermal nests, epidermal consumption, dense dermal nests with atypia. Protective features in the absence of any malignancy indicators were DEJ ring pattern, cobblestone, elongated rete ridges (vertical), well-defined DEJ and wave pattern (vertical). A series of diagnostic criteria for the identification of melanocytic lesions with LC-OCT have been established. Validation of these criteria in clinical practice through future studies is essential to further establish their utility.

Non-Invasive Diagnosis of Angioma Serpiginosum Plantaris: High-Resolution Dermoscopy, High-Frequency Ultrasound, and Line-Field Confocal Optical Coherence Tomography

We present a comprehensive study of acral serpiginous hemangioma, a rare vascular pathology. This distinctive manifestation was examined for the first time using high-resolution video dermoscopy (HRVD), high-frequency ultrasound (HFUS), and line-field confocal optical coherence tomography (LC-OCT). The combined application of these techniques provides new and detailed morphological features able to determine the in vivo structure of lesional skin. This innovative non-invasive approach emphasizes the importance of incorporating advanced diagnostic tools able to provide a virtual histology in order to avoid unnecessary biopsies in benign lesions, often causing pain and functional/aesthetic discomfort.

Spiral scanning improves subject fixation in widefield retinal imaging.

Point scanning retinal imaging modalities, including confocal scanning light ophthalmoscopy (cSLO) and optical coherence tomography, suffer from fixational motion artifacts. Fixation targets, though effective at reducing eye motion, are infeasible in some applications (e.g., handheld devices) due to their bulk and complexity. Here, we report on a cSLO device that scans the retina in a spiral pattern under pseudo-visible illumination, thus collecting image data while simultaneously projecting, into the subject's vision, the image of a bullseye, which acts as a virtual fixation target. An imaging study of 14 young adult volunteers was conducted to compare the fixational performance of this technique to that of raster scanning, with and without a discrete inline fixation target. Image registration was used to quantify subject eye motion; a strip-wise registration method was used for raster scans, and a novel, to the best of our knowledge, ring-based method was used for spiral scans. Results indicate a statistically significant reduction in eye motion by the use of spiral scanning as compared to raster scanning without a fixation target.

An overview of cutaneous squamous cell carcinoma imaging diagnosis methods.

Cutaneous squamous cell carcinoma, a type of non-melanoma skin cancer, is a form of keratinocyte carcinoma that stands as one of the most prevalent cancers, exhibiting a rising frequency. This review provides an overview of the latest literature on imaging methods for diagnosing squamous cell carcinoma (SCC) and actinic keratosis (AK). It discusses the diagnostic criteria, advantages, and disadvantages of various techniques such as dermatoscopy, skin ultrasound (US), in vivo and ex-vivo reflectance confocal microscopy (RCM), and line-field confocal optical coherence tomography (LC-OCT). These methods offer benefits including non-invasiveness, rapidity, comprehensive lesion imaging, and enhanced sensitivity, but face challenges like high costs and the need for specialized expertise. Despite obstacles, the use of these innovative techniques is expected to increase with ongoing technological advancements, improving diagnosis and treatment planning for keratinocyte carcinomas. Standardizing LC-OCT imaging algorithms for AK, Bowen's disease, and SCC remains an area for further research.

Assessment of atopic dermatitis in all phototypes using line‐field confocal optical coherence tomography (LC‐OCT) and reflectance confocal microscopy (RCM)

Assessment of atopic dermatitis in all phototypes using line‐field confocal optical coherence tomography (LC‐OCT) and reflectance confocal microscopy (RCM)

Combining Reflectance Confocal Microscopy, Optical Coherence Tomography and Ex-Vivo Fluorescence Confocal Microscopy for Margin Assessment in Basal Cell Carcinoma Excision.

Recent developments of noninvasive, high-resolution imaging techniques, such as reflectance confocal microscopy (RCM) and optical coherence tomography (OCT), have enhanced skin cancer detection and precise tumor excision particularly in highly aggressive and poorly defined basal cell carcinomas (BCCs). The aim of this pilot study is to assess the feasibility and reproducibility of a systematic clinical workflow combining noninvasive (RCM-OCT) and invasive fluorescence confocal microscopy (FCM) imaging modalities in pre- and intra-surgical evaluations of the lateral and deep margins of BCC. Superficial incisions were made 2 mm beyond the clinical-dermoscopic BCC margins. Lateral margins were then explored with OCT and RCM. In positive margins, a further cut was made 2 mm distal from the previous. A final RCM/OCT-based double-negative margin was drawn around the entire perimeter of the lesion before referring to surgery. The freshly excised specimen was then examined with FCM (ex-vivo) for the evaluation of the deep margin. Histopathologic examination eventually confirmed margin involvement. The study included 22 lesions from 13 patients. At the end of the study, 146 margins-106 negative (73%) and 40 positive (27%) at RCM/OCT-were collected. The RCM/OCT margin evaluation showed an overall sensitivity of 100% and a specificity of 96.3%. The overall positive margins diagnostic accuracy was 98.2%. Reproducibility was evaluated on recorded images and the raters showed a substantial inter-observer agreement on both RCM (κ = 0.752) and OCT images (κ = 0.724). The combined RCM/OCT/FCM ex-vivo approach noninvasively facilitates the presurgical and intrasurgical lateral and deep margin assessment of poorly defined BCCs.

Line-field confocal optical coherence tomography in dermato-oncology: A literature review towards harmonized histopathology-integrated terminology.

Non-invasive diagnostics like line-field confocal optical coherence tomography (LC-OCT) are being implemented in dermato-oncology. However, unification of terminology in LC-OCT is lacking. By reviewing the LC-OCT literature in the field of dermato-oncology, this study aimed to develop a unified terminological glossary integrated with traditional histopathology. A PRISMA-guided literature-search was conducted for English-language publications on LC-OCT of actinic keratosis (AK), keratinocyte carcinoma (KC), and malignant melanoma (MM). Study characteristics and terminology were compiled. To harmonize LC-OCT terminology and integrate with histopathology, synonymous terms for image features of AK, KC, and MM were merged by two authors, organized by skin layer and lesion-type. A subset of key LC-OCT image-markers with histopathological correlates that in combination were typical of AK, squamous cell carcinoma insitu (SCCis), invasive squamous cell carcinoma (SCC), basal cell carcinoma (BCC), and MM in traditional histopathology, were selected from the glossary by an experienced dermatopathologist. Seventeen observational studies of AK (7 studies), KC (13 studies), MM (7 studies) utilizing LC-OCT were included, with 117 terms describing either AK, KC, or MM. These were merged to produce 45 merged-terms (61.5% reduction); 5 assigned to the stratum corneum (SC), 23 to the viable epidermis, 2 to dermo-epidermal junction (DEJ) and 15 to the dermis. For each lesion, mandatory key image-markers were a well-defined DEJ and presence of mild/moderate but not severe epidermal dysplasia for AK, severe epidermal dysplasia and well-defined DEJ for SCCis, interrupted DEJ and/or dermal broad infiltrative strands for invasive SCC, dermal lobules connected and/or unconnected to the epidermis for BCC, as well as single atypical melanocytes and/or nest of atypical melanocytes in the epidermis or dermis for MM. This review compiles evidence on LC-OCT in dermato-oncology, providing a harmonized histopathology-integrated terminology and key image-markers for each lesion. Further evaluation is required to determine the clinical value of these findings.

In vivo assessment of early effects of diamond-tipped microdermabrasion through the lens of line-field confocal optical coherence tomography.

Microdermabrasion is a cosmetic procedure that has gained popularity for skin rejuvenation by causing repetitive intraepidermal injury to stimulate the proliferation of fibroblasts and collagen production. Various clinical studies have demonstrated microdermabrasion's effectiveness in skin rejuvenation; however, most of these studies rely on clinical observation and scoring by observers rather than histologic or microscopic analysis. In our single-center prospective study, we used line-field confocal optical coherence tomography (LC-OCT), to non-invasively visualize the early effects of one microdermabrasion treatment on the facial epidermal and dermal structure. Using LC-OCT, this study aims to elucidate the microscopic and histological effects of microdermabrasion on epidermal and dermal structures, including epidermal thickness, as well as collagen and vascular patterns. Eight volunteers (Fitzpatrick skin types II-V) underwent one treatment of microdermabrasion. LC-OCT and VISIA imaging were performed before and 10 min after microdermabrasion, and at 48-h follow-up. Subjective evaluations of skin texture and adverse reactions were assessed 1 week posttreatment via a telephone call. Compared to LC-OCT images before treatment, images captured after one treatment of microdermabrasion showed a decrease in thickness and number of undulations in the stratum corneum. In the superficial dermis, enhancement in fibrillar collagen, as demonstrated by an increased prominence of crisscrossing hyper-refractile strands, was visualized. This was consistent with subjective and objective improvement in facial rhytids calculated by VISIA skin analysis. Treatment monitoring with LC-OCT demonstrated consistent histopathological changes with clinical visual improvement. Therefore, LC-OCT, has the potential to enable long-term histopathological monitoring of microdermabrasion and other cosmetic procedures without biopsy.

Effects of refractive index mismatch between sample and immersion medium in line-field confocal optical coherence tomography

Line-field confocal optical coherence tomography (LC-OCT) is an optical technique based on low-coherence interference microscopy with line illumination, designed for tomographic imaging of semi-transparent samples with micrometer-scale spatial resolution. A theoretical model of the signal acquired in LC-OCT is presented. The model shows that a refractive index mismatch between the sample and the immersion medium causes a dissociation of the coherence plane and the focal plane, leading to a decrease in the signal amplitude and a degradation of the image’s lateral resolution. Measurements are performed to validate and illustrate the theoretical predictions. A mathematical condition linking various experimental parameters is established to ensure that the degradation of image quality is negligible. This condition is tested experimentally by imaging a phantom. It is verified theoretically in the case of skin imaging, using experimental parameters corresponding to those of the commercially available LC-OCT device.

Features of tattoo-associated cutaneous lymphoid hyperplasia on reflectance confocal microscopy and line-field confocal optical coherence tomography.

The popularity of tattoos has led to an increase in associated skin reactions, including complications such as infection, allergic reactions and rare conditions such as tattoo-induced cutaneous lymphoid hyperplasia (CLH). CLH is a benign lymphoproliferative reaction with clinical features resembling malignant cutaneous lymphomas. Non-invasive diagnostic tools like reflectance confocal microscopy (RCM) and the new line-field confocal optical coherence tomography (LC-OCT) are being studied in dermatology better to understand the morphological patterns of many dermatological diseases. Between September 2021 and May 2023, patients with suspicious lesions for tattoo-related CLH were analysed using RCM and LC-OCT before confirming the diagnosis of CLH through skin biopsy and histopathological examination. The study included five cases of CLH. It focused on the analysis of high-quality LC-OCT images/videos and RCM images to investigate the features of CLH in tattooed individuals. Most (80%) cases exhibited a mixed T and B lymphocyte infiltration subtype, while 20% showed a predominant T infiltration subtype. RCM and LC-OCT revealed characteristic features, including architectural disarray, fibrosis, lymphoid infiltrates, and pigment deposits in the epidermis and dermis. Non-invasive tools such as RCM and LC-OCT are valuable in diagnosing tattoo-related CLH. While skin biopsy remains the current standard for diagnosis, RCM and LC-OCT can serve as helpful adjuncts in identifying the most representative area for biopsy. They may potentially become alternative diagnostic options in the future, offering benefits in terms of cost, diagnostic efficiency, aesthetics and patient satisfaction as the prevalence of tattoo-related adverse reactions continues to rise.

A Novel Flexible Endomicroscopic Scanning Instrument with Visual Servoing Control

Optical biopsy using probe‐based confocal laser endomicroscopy (pCLE) and optical coherence tomography (OCT) technologies performs in vivo tissue scanning of organ surfaces and plays a crucial role in early cancer diagnosis. Herein, a flexible endomicroscopic scanning instrument based on a 3‐degree‐of‐freedom flexible hinge structure and visual servoing control is presented. The instrument integrates OCT and pCLE. OCT information compensates for the imaging distance of pCLE, which can combine the advantages of the two technologies. The mechanism allows for tissue surface scanning with a wire‐driven active bending part and dynamic adjustment of the probe‐tissue distance using a combination of wire and spring‐driven, which satisfy large‐area scanning while maintaining consistence tissue contacts. Local optimization‐based visual servoing control is proposed to optimize the tissue scanning. Through the scanning experiments, high‐quality mosaics of uneven surfaces are obtained with an effective area that is larger than 4 mm2. In vivo animal trial further verifies the feasibility and clinical potential of the instrument.

Combined reflectance confocal microscopy and optical coherence tomography in the evaluation of cutaneous squamous cell carcinoma

Combined reflectance confocal microscopy and optical coherence tomography in the evaluation of cutaneous squamous cell carcinoma

Diagnostic Imaging of Agminated Blue Lesions and Blue Lesions with Satellitosis: Case Series with a Concise Review of the Current Literature

Background: Agmination and/or satellitosis in pigmented blue lesions is a phenomenon rarely mentioned in the literature and not well known. This phenomenon can be expressed by several benign and malignant pigmented blue lesions, such as blue nevi, Spitz nevi, melanocytoma and melanoma. On this spectrum, dermoscopy, reflectance confocal microscopy (RCM) and dynamic Optical coherence tomography (D-OCT) represent non-invasive imaging technologies, which may help clinicians in the diagnosis of melanoma and non-melanoma skin cancers in daily clinical practice. Methods: Currently, in the literature there is a lack of new data about agminated blue lesions and blues lesions with satellitosis, as well as the lack of a recent and updated review of the literature about this topic. Therefore, considering that clinicians must be confident with the diagnosis of these rare skin lesions, we decided to carry out this work. Results: In this paper, four new cases of agminated pigmented cutaneous lesions were described. Moreover, a review of the current literature on this topic was performed. Conclusions: A clinical–pathological correlation is often needed to reach a correct diagnosis; currently, dermoscopy and non-invasive diagnostic techniques, such as reflectance confocal microscopy and optical coherence tomography, due to the depth of these skin lesions in the dermis, can only make a partial and limited contribution.

Skin dark spot mapping and evaluation of brightening product efficacy using Line-field Confocal Optical Coherence Tomography (LC-OCT).

Facial dark spots remain a significant challenge for the cosmetic industry, in terms of providing effective treatment. Using Line-field Confocal Optical Coherence Tomography (LC-OCT), we investigated the internal structural features of photo-aging spot areas and evaluated the efficacy of a skin-brightening cosmetic product. Twenty-six Asian female volunteers, aged between 29 and 65 years, applied a cosmetic product on their entire face twice a day for 2 months. LC-OCT was used to evaluate the dermal-epidermal junction (DEJ) undulation and the volume density of melanin in the epidermis at D0 and D56. Skin brightening and redness were also assessed by photography (SkinCam). Using LC-OCT technology, various microscopic dark spot morphologies, spanning from minimally deformed DEJ to complex DEJ patterns, were identified. Dark spots characterized by slight deformities in the DEJ were predominantly observed in the youngest age group, while older volunteers displayed a wavier pattern. Furthermore, a total of 44 spots were monitored to evaluate the brightening product efficacy. A statistically significant reduction in melanin volumetric density of 7.3% in the spots and 12.3% in their surrounding area was observed after 56 days of product application. In line with these results, an analysis of color parameters using SkinCam reveals a significant increase in brightening and decrease in redness in both pigmented spots and the surrounding skin following application. LC-OCT proves to be a valuable tool for in-depth dark spots characterization and assessment of skin brightening products, enabling various applications in the field of dermatological sciences.

Line-field confocal optical coherence tomography, a novel non-invasive tool for the diagnosis of onychomycosis.

Onychomycosis is common and important to distinguish from other nail diseases. Rapid and accurate diagnosis is necessary for optimal patient treatment and outcome. Non-invasive diagnostic tools have increasing potential for nail diseases including onychomycosis. This study evaluated line-field confocal optical coherence tomography (LC-OCT) as a rapid non-invasive tool for diagnosing onychomycosis as compared to confocal laser scanning microscopy (CLSM), optical coherence tomography (OCT), and conventional methods. In this prospective study 86 patients with clinically suspected onychomycosis and 14 controls were examined using LC-OCT, OCT, and CLSM. KOH-preparation, fungal culture, PCR, and histopathology were used as comparative conventional methods. LC-OCT had the highest sensitivity and negative predictive value of all methods used, closely followed by PCR and OCT. Specificity and positive predictive value of LC-OCT were as high as with CLSM, while OCT scored much lower. The gold standard technique, fungal culture, showed the lowest sensitivity and negative predictive value. Only PCR and culture allowed species differentiation. LC-OCT enables quick and non-invasive detection of onychomycosis, with advantages over CLSM and OCT, and similar diagnostic accuracy to PCR but lacking species differentiation. For accurate nail examination, LC-OCT requires well-trained and experienced operators.

3D Visualization of cyanobacterial biofilms using micro-computed tomography with contrast-enhancing staining agents

Currently, biofilms colonizing surfaces are mainly imaged in 2D by conventional techniques, such as optical or scanning electron microscopy. Confocal laser scanning microscopy or optical coherence tomography can visualize biofilms in 3D, but they suffer from a limited penetration depth and cannot visualize biofilms in opaque materials. Micro-computed tomography (µCT) can overcome these issues, but µCT cannot easily distinguish biofilm structures from water due to a lack of contrast difference. Within this research, five contrast-enhancing staining agents (CESAs) were evaluated for their staining potential of cyanobacterial biofilms, aiming to visualize these biofilms in 3D. Isotonic Lugol and 1:2 hafnium(IV)-substituted Wells-Dawson polyoxometalate (Hf-WD 1:2 POM) were the most promising, as they allowed visualization of the biofilms and revealed structures in the stained biofilms. Staining with isotonic Lugol could clearly visualize bundles of filaments within the biofilm, while Hf-WD 1:2 POM revealed a smooth biofilm. It is assumed that both CESAs have a different affinity towards the biofilms and could thus be used complementary. Monolacunary Wells-Dawson polyoxometalate (Mono-WD POM) showed moderate discrimination while staining with cationic iodinated CA4+ and Hexabrix® (Guerbet) containing anionic ioxaglate did not allow to distinctly visualize the biofilms. These results indicate that µCT, together with CESAs such as isotonic Lugol and Hf-WD 1:2 POM, can be used as a tool to image extensive biofilms or microbial mats in 3D. Further research will determine whether these CESAs are suitable for visualizing biofilms within opaque porous media.

From Clinical Suspicion to Diagnosis: A Review of Diagnostic Approaches and Challenges in Fungal Keratitis.

Fungal keratitis is a relatively rare yet severe ocular infection that can lead to profound vision impairment and even permanent vision loss. Rapid and accurate diagnosis plays a crucial role in the effective management of the disease. A patient's history establishes the initial clinical suspicion since it can provide valuable clues to potential predisposing factors and sources of fungal exposure. Regarding the evaluation of the observed symptoms, they are not exclusive to fungal keratitis, but their timeline can aid in distinguishing fungal keratitis from other conditions. Thorough clinical examination of the affected eye with a slit-lamp microscope guides diagnosis because some clinical features are valuable predictors of fungal keratitis. Definitive diagnosis is established through appropriate microbiological investigations. Direct microscopic examination of corneal scrapings or biopsy specimens can assist in the presumptive diagnosis of fungal keratitis, but culture remains the gold standard for diagnosing fungal keratitis. Advanced molecular techniques such as PCR and MALDI-ToF MS are explored for their rapid and sensitive diagnostic capabilities. Non-invasive techniques like in vivo confocal microscopy (IVCM) and optical coherence tomography (OCT) are useful for real-time imaging. Every diagnostic technique has both advantages and drawbacks. Also, the selection of a diagnostic approach can depend on various factors, including the specific clinical context, the availability of resources, and the proficiency of healthcare personnel.

Frequency selection rule for the HDHF Lissajous scanning imaging with a low-voltage one axis actuated PZT scanner based on an asymmetric fiber cantilever.

Lissajous micro scanners are very attractive in compact laser scanning applications for biomedical endoscopic imaging, such as confocal microscopy, endomicroscopy or optical coherence tomography. The scanning frequencies have a very important effect on the quality of the resulting Lissajous scanning imaging. In this paper, we propose a frequency selection rule for high definition and high frame-rate (HDHF) Lissajous scanning imaging, by deriving the relationship among the scanning field of view (FOV), actuation frequencies and pixel size based on the characteristics of the scanning trajectory. The minimum sampling rate based on the proposed frequency selection rule is further discussed. We report a lead zirconate titanate piezoelectric ceramic (PZT) based Lissajous fiber scanner to achieve HDHF Lissajous scanning imaging. Based on the frequency selection rule, different frequency combinations are calculated, under which the Lissajous fiber scanner can work at the frame rate (FR) of 10 Hz, 20 Hz, 40 Hz and 52 Hz. The trajectory evolution of the Lissajous scanning at the frame rate of 10 Hz has been obtained to verify the applicability of the proposed rule. The measured resolution of the scanner is 50.8 lp/mm at the unit optical magnification, and the measured FOV at the FR of 10 Hz and 40 Hz are 1.620 mm ×1.095 mm and 0.405 mm ×0.27 mm, respectively. HDHF Lissajous scanning images of the customized spatial varying binary pattern are obtained and reconstructed at the FR of 10 Hz and 40 Hz, demonstrating the practicability of the frequency selection rule.