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Introduction to reflectance confocal microscopy and its use in clinical practice
摘要： Re?ectance confocal microscopy (RCM) is a novel technology that provides noninvasive, in vivo imaging of the skin at nearly histologic resolution. In 2016, the US Centers for Medicare and Medicaid Services (CMS) established reimbursement codes for RCM image acquisition and for the reading and interpretation of images. The combination of RCM imaging with dermoscopy has improved the accuracy of skin cancer diagnosis while reducing the number of biopsies of benign skin lesions. With that, we are starting to see more dermatologists and dermatopathologists using RCM in clinical practice. This editorial is to serve as an introduction on RCM imaging with a focus on its usefulness in both the diagnosis and management of skin cancers. We end by brie?y describing the characteristic RCM features of normal skin to serve as a building block for later cases that will explore both the bene?ts and drawbacks of incorporating RCM imaging for benign and malignant lesions.
关键词： innovative technology,noninvasive imaging,skin cancer,melanoma,re?ectance confocal microscopy,lentigo maligna,nonmelanoma skin cancer
更新于2025-09-23 15:21:01
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Laser Scanning Confocal Microscopy 3D Surface Metrology Applications
摘要： Surface metrology, measurement of solid surfaces topography, has become an important topic for many material scientists and engineers. Characterization of the surfaces can help researchers find new functional materials, improve device performance and so forth. ‘Seeing is believing.’ Visualization of fine 3-dimensional (3D) details of surfaces is critical in surface metrology studies. Among various observation and measurement techniques, Laser Scanning Confocal Microscopy (LSCM) becomes more and more popular due to the fact that it is non-contact and non-destructive to the samples, requires minimal sample preparation, and efficient automations and provides single nanometer level resolution. In this paper, we will introduce these features in detail, by using Olympus LEXT OLS5000, the newest 3D laser confocal scanning microscope.
关键词： Olympus LEXT OLS5000,3D Surface Metrology,Laser Scanning Confocal Microscopy
更新于2025-09-23 15:19:57
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Rapid identification of human mast cell degranulation regulators using functional genomics coupled to high-resolution confocal microscopy
摘要： Targeted functional genomics represents a powerful approach for studying gene function in vivo and in vitro. However, its application to gene expression studies in human mast cells has been hampered by low yields of human mast cell cultures and their poor transfection efficiency. We developed an imaging system in which mast cell degranulation can be visualized in single cells subjected to shRNA knockdown or CRISPR–Cas9 gene editing. By using high-resolution confocal microscopy and a fluorochrome-labeled avidin probe, one can directly assess the alteration of functional responses, i.e., degranulation, in single human mast cells (10–12 weeks old). The elimination of a drug or marker selection step avoids the use of potentially toxic treatment procedures, and the brief hands-on time of the functional analysis step enables high-throughput screening of shRNA or CRISPR–Cas9 constructs to identify genes that regulate human mast cell degranulation. The ability to analyze single cells substantially reduces the total number of cells required and enables the parallel visualization of the degranulation profiles of both edited and non-edited mast cells, offering a consistent internal control not found in other protocols. Moreover, our protocol offers a flexible choice between RNA interference (RNAi) and CRISPR–Cas9 genome editing for perturbation of gene expression using our human mast cell single-cell imaging system. Perturbation of gene expression, acquisition of microscopy data and image analysis can be completed within 5 d, requiring only standard laboratory equipment and expertise.
关键词： confocal microscopy,human mast cells,CRISPR–Cas9,degranulation,shRNA,functional genomics
更新于2025-09-23 15:19:57
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Effect of galvanotaxic graphene oxide on chloroplast activity: Interaction quantified with Biolayer-Interferometry coupled confocal microscopy
摘要： The knowledge on the effect of graphene oxide (GO) on plants are limited to germination, growth, and toxicity. Since, chloroplast generates sugar by the reduction of CO2 with the optical stimulation through a series of electron transport chain and GO being the 2D material with electron transport property, it is reasonable to check their interaction. Here, the effect of GO without and with amine conjugation (AGO) having opposite charges were allowed to interact with chloroplast. The uptake is documented by using biolayer interferometry coupled with confocal imaging. The ex vivo chloroplast activity with GO and AGO has been tested and found that the GO treatment shows 1.3 times more activity than control. In contrast, AGO function as efficient electron conductor and cause imbalance in the redox beyond the capacity of the antioxidant rich chloroplast solute. Finally, in vivo toxicity has been evaluated in the spinach plants, which highlights the chance of AGO application as herbicide to remove any unwanted plants.
关键词： Graphene oxide,Confocal microscopy,Herbicide,Biolayer-Interferometry,Chloroplast activity
更新于2025-09-23 15:19:57
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Versatile high-speed confocal microscopy using a single laser beam
摘要： We present a new flexible high speed laser scanning confocal microscope and its extension by an astigmatism particle tracking velocimetry (APTV) device. Many standard confocal microscopes use either a single laser beam to scan the sample at a relatively low overall frame rate or many laser beams to simultaneously scan the sample and achieve a high overall frame rate. The single-laser-beam confocal microscope often uses a point detector to acquire the image. To achieve high overall frame rates, we use, next to the standard 2D probe scanning unit, a second 2D scan unit projecting the image directly onto a 2D CCD-sensor (re-scan configuration). Using only a single laser beam eliminates crosstalk and leads to an imaging quality that is independent of the frame rate with a lateral resolution of 0.235 μm. The design described here is suitable for a high frame rate, i.e., for frame rates well above the video rate (full frame) up to a line rate of 32 kHz. The dwell time of the laser focus on any spot in the sample (122 ns) is significantly shorter than those in standard confocal microscopes (in the order of milli- or microseconds). This short dwell time reduces phototoxicity and bleaching of fluorescent molecules. The new design opens up further flexibility and facilitates coupling to other optical methods. The setup can easily be extended by an APTV device to measure three dimensional dynamics while being able to show high resolution confocal structures. Thus, one can use the high resolution confocal information synchronized with an APTV dataset.
关键词： APTV,high-speed imaging,astigmatism particle tracking velocimetry,confocal microscopy,single laser beam
更新于2025-09-23 15:19:57
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In vivo confocal microscopy classification in the diagnosis of meibomian gland dysfunction
摘要： Aim Meibomian gland dysfunction (MGD) is one of the most common disorders in ophthalmology. The aim of this study was to evaluate the use of this in vivo confocal microscopy (IVCM)-MGD description to classify patients affected by clinical MGD and measure the correlation with standard clinical criteria and subjective symptoms. Methods One hundred eyes of 100 patients suffering from MGD and 15 eyes of normal subjects were included. A comprehensive evaluation with the ocular surface disease index (OSDI), Schirmer test, tear break-up time (TBUT), tear osmolarity, Oxford score, Meibomian gland expression, palpebral IVCM, and meibography was performed. Then each patient was classi?ed using a new IVCM classi?cation: type 0 for normality, type 1 for meibum obstruction, type 2 for in?ammation, and type 3 for ?brosis. Results The mean age of patients was 52 ± 20 years old, the OSDI was 38 ± 23, the BUT 5 ± 2.6 s, the Schirmer test 13 ± 7 mm, tear osmolarity 300 ± 11 osmol/L, the Oxford score 0.5 ± 0.6, the meibum expression score 1.7 ± 1.02, and the meibography score 1.3 ± 0.9. The IVCM MG classi?cation of the 15 normal subjects was 0. For MGD patients, 29% were in type 1, 40% were type 2, and 31% were type 3. The patients in IVCM MG type 2 had a higher OSDI (p = 0.001) compared with the other types. There was a strong correlation between the IVCM score and the meibography score (r = 0.71 p < 0.0001). Conclusion This new IVCM classi?cation provided a practical pathophysiological system for MGD. By giving objective criteria, this IVCM classi?cation may help advance the understanding of patients’ symptoms and enhance treatment effectiveness in MGD.
关键词： classification,Meibomian gland dysfunction,diagnosis,in vivo confocal microscopy
更新于2025-09-23 15:19:57
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Apport de la microscopie confocale par réflectance et de l’OCT dans le diagnostic d’un carcinome basocellulaire nodulaire kystique
摘要： The role of reflectance confocal microscopy and OCT in the diagnosis of nodular kystic BCC
关键词： Basal cell carcinoma,Noninvasive imaging,Optical coherence tomography,In vivo confocal microscopy,Dermoscopy,In vivo reflectance confocal microscopy,Cystic basal cell carcinoma
更新于2025-09-19 17:15:36
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In Vivo Corneal Confocal Microscopy: Pre- and Post-operative Evaluation in a Case of Corneal Neurotization
摘要： In this report, we analyse the pre- and post-operative corneal changes observed using in vivo confocal corneal microscopy in a patient with neurotrophic keratitis submitted to a corneal reinnervation surgical procedure. We describe favourable trophic changes observed at different levels of the patient’s cornea, particularly in the sub-basal nerve plexus; complete absence of these neurological structures was observed before surgery, but appeared largely restored six months thereafter.
关键词： Corneal confocal microscopy,corneal neurotization,neurotrophic keratitis
更新于2025-09-19 17:15:36
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Confidence Distance Matrix for outlier identification: A new method to improve the characterizations of surfaces measured by confocal microscopy
摘要： This paper proposes a statistical method for outlier identification for surface measurement data obtained by confocal microscopy. The implemented statistical method is Confidence Distance Matrix (CDM) which were widely used in statistics and many engineering areas, such as signal processing, sensor data fusion, information problems, etc. However, no investigations on identifying outliers in measured surface data using CDM have been found. This paper introduces and simplifies the mathematical model of CDM method. Algorithms for identifying random outliers using Monte Carlo method for uncertainty evaluation and for identifying outliers in a unique measured surface are developed and validated. For validation of the algorithms, a synthetic data SG_3-3 provided by National Institute of Standards and Technology and a data of artificial stochastic surface generated by our own algorithms are implemented. The difference of Sq of the data with outliers is 2.3342% and after deletion of outliers is 0.0037% with reference to the certified value. A type C1 spacing standard with dust dropped is measured and processed using CDM. The difference of Sa decreases from 29.65% to 3.52% after processing outliers with reference to the certified value Ra. An iron surface is measured and processed. Surface slopes and curvatures of the data in the two validations and two experiments are compared. All those parameters, the surface reconstructions, histogram of heights, and QQ plot of the measured surface data versus the data after deletion of outliers indicate our proposed method working well.
关键词： Confidence Distance matrix,outlier detection,areal surface characterization,threshold determination,Imaging Confocal Microscopy,Monte Carlo method
更新于2025-09-19 17:15:36
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Microneedle fractional radiofrequency-induced micropores evaluated by in vivo reflectance confocal microscopy, optical coherence tomography, and histology
摘要： Background: Microneedle fractional radiofrequency (MNRF) is a minimally invasive technique that delivers radiofrequency (RF) energy into the skin via microneedles. Reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) enable the characterization of device-tissue interactions in in vivo skin. The aim of this study is to describe MNRF-induced micropores using RCM and OCT imaging. Materials and methods: Five healthy participants were treated with a 7 × 7 array of 1500 μm microneedles on two adjacent areas of the right hip. One area received MNRF using high RF energy while the other underwent MNRF at low RF energy. Micropore morphology was evaluated qualitatively and quantitatively with RCM and OCT. To relate imaging with histology, one participant underwent punch biopsy in both areas. Results: Reflectance confocal microscopy visualized shape, content, and thermal-induced coagulation zone (CZ) of MNRF micropores. At high RF energy, micropores showed concentric shape, contained hyperreflective granules, and coagulated tissue from epidermis to dermo-epidermal junction (diameter 63-85 μm). Micropores at low RF energy, presented with a stellate shape, no content and CZs that were visible only in epidermis (CZ thickness 9 μm, IQR 8-21 μm). Evaluating OCT, high RF energy showed deeper (150 μm), more easily identifiable micropores compared to low RF energy micropores (70 μm). Histology showed tissue coagulation to a depth of 1500 μm at high RF energy, while at low RF energy, disruption was only visible in epidermis. Conclusion: Microneedle fractional radiofrequency micropores show distinct characteristics in both RCM and OCT, depending on RF energy. These in vivo imaging modalities are complementary and allow combined, qualitative, and quantitative evaluation.
关键词： microneedle fractional radiofrequency,histology,optical coherence tomography,micropore,reflectance confocal microscopy
更新于2025-09-19 17:15:36