- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Image and data processing algorithms for identifying cell-bound membrane vesicle trajectories and movement information
摘要： This DIB article provides details about the trajectory identification and data processing algorithms used in the article "Dynamic single-vesicle tracking of cell-bound membrane vesicles on resting, activated, and cytoskeleton-disrupted cells" (Zhang et al.) [1]. The algorithm identifies vesicles on cell membranes from series of undyed grayscale images captured by the confocal microscope based on contrast differences and then trajectories of vesicles are obtained by analyzing their positions in consecutive images. Once the trajectories have been obtained, other quantitative movement information, such as moving speed, direction and acceleration, are derived by standard dynamic relations.
关键词： MATLAB algorithms,Vesicle tracking,Trajectory tracing,Confocal microscopy,Image processing
更新于2025-09-23 15:22:29
-
Quantitative imaging of membrane micropolarity in living cells and tissues by spectral phasors analysis
摘要： Intracellular micropolarity is essential in several metabolic processes, as it controls membrane permeability, fluxes of molecules and energy. Here we describe a method for the determination of the micropolarity in living cells using spectral confocal microscopy. The method is based on a phasor analysis of spectrally resolved images of live cells, labelled with the solvatochromic probe Nile Red. An application is provided to extract a polarity profile from the acquired Spectral datasets, which represent the contribution of hyperpolar, polar and non-polar lipids, and to generate a micropolarity map at submicrometric spatial resolution. A metabolic parameter, representing a quantitative index of the fatty acid-triacylglycerol turnover, is also furnished. This method allows a functional profiling of cells and tissues and the detection of metabolic imbalances between lipid storage and usage.
关键词： Membranes micropolarity,Nile Red,Fatty acids,Triglycerides,Lipid droplets,Confocal microscopy,Metabolic imaging,Spectral phasors,Lipids
更新于2025-09-23 15:22:29
-
Reflectance confocal microscopy of cutaneous melanoma. Correlation with dermoscopy and histopathology
摘要： In vivo Confocal Microscopy is a method for non-invasive, real-time visualization of microscopic structures and cellular details of the epidermis and dermis, which has a degree of resolution similar to that obtained with histology. We present a case of cutaneous melanoma in which diagnosis was aided by confocal microscopy examination. We also correlate the observed features with the dermoscopic and histopathological findings. Confocal microscopy proved to be an useful adjunct to dermoscopy, playing an important role as a method 'between clinical evaluation and histopathology'.
关键词： Melanoma,Diagnosis,Dermoscopy,Confocal microscopy
更新于2025-09-23 15:22:29
-
Cutaneous erysipeloid metastasis of cholangiocarcinoma and evaluation by in?vivo reflectance confocal microscopy
摘要： Cutaneous metastases are relatively uncommon, occurring in only 0.7% to 9% of all internal malignancies. Cholangiocarcinoma is a rare bile duct neoplasm that accounts for less than 2% of malignancies. Although it is well known that cholangiocarcinoma metastasizes to the lungs, liver, peritoneum, and retroperitoneal lymph nodes, a retrospective review of the literature from 1978 to 2014 indicates only 30 cases of cutaneous cholangiocarcinoma, with 17 cases presenting without concurrent metastasis in other sites. Among these patients, the cutaneous metastatic disease occurred evenly at adjacent and distant sites, presenting as 0.3-cm to 4-cm erythematous papules or nodules with or without ulceration. The median overall survival after diagnosis of cutaneous cholangiocarcinoma metastasis is 4 months. Paradoxically, single-site metastases carry a significantly worse prognosis than multiple-site metastases and may be attributed to difficulty in identifying a singular lesion. In vivo reflectance confocal microscopy (RCM) is a novel, noninvasive diagnostic alternative to skin biopsy with comparable insurance reimbursement that captures real-time, high-resolution, cellular-level images from the skin surface down to the reticular dermis (up to 300 μm depth). This modality forgoes traumatic biopsy and has been used for diagnosis and monitoring of skin cancers and inflammatory dermatoses.
关键词： cholangiocarcinoma,erysipeloid,cutaneous metastasis,reflectance confocal microscopy
更新于2025-09-23 15:21:21
-
Corneal Confocal Microscopy detects a Reduction in Corneal Endothelial Cells and Nerve Fibres in Patients with Acute Ischemic Stroke
摘要： Endothelial dysfunction and damage underlie cerebrovascular disease and ischemic stroke. We undertook corneal confocal microscopy (CCM) to quantify corneal endothelial cell and nerve morphology in 146 patients with an acute ischemic stroke and 18 age-matched healthy control participants. Corneal endothelial cell density was lower (P < 0.001) and endothelial cell area (P < 0.001) and perimeter (P < 0.001) were higher, whilst corneal nerve fibre density (P < 0.001), corneal nerve branch density (P < 0.001) and corneal nerve fibre length (P = 0.001) were lower in patients with acute ischemic stroke compared to controls. Corneal endothelial cell density, cell area and cell perimeter correlated with corneal nerve fiber density (P = 0.033, P = 0.014, P = 0.011) and length (P = 0.017, P = 0.013, P = 0.008), respectively. Multiple linear regression analysis showed a significant independent association between corneal endothelial cell density, area and perimeter with acute ischemic stroke and triglycerides. CCM is a rapid non-invasive ophthalmic imaging technique, which could be used to identify patients at risk of acute ischemic stroke.
关键词： Corneal Confocal Microscopy,corneal endothelial cells,acute ischemic stroke,corneal nerve fibres
更新于2025-09-23 15:21:21
-
[Methods in Molecular Biology] Barley Volume 1900 (Methods and Protocols) || Preparation of Barley Pollen Mother Cells for Confocal and Super Resolution Microscopy
摘要： Recombination (crossover) drives the release of genetic diversity in plant breeding programs. However, in barley, recombination is skewed toward the telomeric ends of its seven chromosomes, restricting the re-assortment of about 30% of the genes located in the centromeric regions of its large 5.1 Gb genome. A better understanding of meiosis and recombination could provide ways of modulating crossover distribution and frequency in barley as well as in other grasses, including wheat. While most research on recombination has been carried out in the model plant Arabidopsis thaliana, recent studies in barley (Hordeum Vulgare) have provided new insights into the control of crossing over in large genome species. A major achievement in these studies has been the use of cytological procedures to follow meiotic events. This protocol provides detailed practical steps required to perform immunostaining of barley meiocytes (pollen mother cells) for confocal or structured illumination microscopy.
关键词： Antibodies,Barley,3D-SIM,Immuno-cytochemistry,Confocal Microscopy,Meiosis
更新于2025-09-23 15:21:21
-
Imaging Supramolecular Morphogenesis with Confocal Laser Scanning Microscopy at Elevated Temperatures
摘要： The morphogenesis of supramolecular assemblies is a highly dynamic process that has only recently been recognized, and our understanding of this phenomenon will require imaging techniques capable of crossing scales. Shape transformations depend both on the complex energy landscapes of supramolecular systems and the kinetically controlled pathways that define their structures and functions. We report here the use of confocal laser scanning microscopy coupled with a custom-designed variable-temperature sample stage that enables in situ observation of such shape changes. The submicrometer resolution of this technique allows for real-time observation of the nanostructures in the native liquid environments in which they transform with thermal energy. We use this technique to study the temperature-dependent morphogenic behavior of peptide amphiphile nanofibers and photocatalytic chromophore amphiphile nanoribbons. The variable-temperature confocal microscopy technique demonstrated in this work can sample a large volume and provides real-time information on thermally induced morphological changes in the solution.
关键词： in situ microscopy,confocal microscopy,nanofibers,nanoribbons,supramolecular assembly
更新于2025-09-23 15:21:01
-
Increasing fluorescence lifetime for resolution improvement in STED nanoscopy
摘要： Super-resolution microscopy (SRM) has had a substantial impact on the biological sciences due to its ability to observe tiny objects less than 200 nm in size. Stimulated emission depletion (STED) microscopy represents a major category of these SRM techniques that can achieve diffraction-unlimited resolution based on a purely optical modulation of fluorescence behaviors. Here, we investigated how the laser beams affect fluorescence lifetime in both confocal and STED imaging modes. The results showed that with increasing illumination time, the fluorescence lifetime in two kinds of fluorescent microspheres had an obvious change in STED imaging mode, compared that in confocal imaging mode. As a result, the reduction of saturation intensity induced by the increase of fluorescence lifetime can improve the STED imaging resolution at the same depletion power. The phenomenon was also observed in Star635P labeled human Nup153 in fixed HeLa cells, which can be treated as a reference for the synthesis of fluorescent labels with the sensitivity to the surrounding environment for resolution improvement in STED nanoscopy.
关键词： confocal microscopy,super-resolution,fluorescence lifetime,fluorescence microscopy
更新于2025-09-23 15:21:01
-
Laser Scanning Confocal Microscopy Was Used to Validate the Presence of Burkholderia pseudomallei or B. mallei in Formalin-Fixed Paraffin Embedded Tissues
摘要： Burkholderia pseudomallei and B. mallei are Gram-negative, facultative intracellular bacteria that cause melioidosis and glanders, respectively. Currently, there are no vaccines for these two diseases. Animal models have been developed to evaluate vaccines and therapeutics. Tissues from infected animals, however, must be fixed in formalin and embedded in paraffin (FFPE) before analysis. A brownish staining material in infected tissues that represents the exopolysaccharide of the pathogen was seen by bright field microscopy but not the actual microorganism. Because of these results, FFPE tissue was examined by laser scanning confocal microscopy (LSCM) in an attempt to see the microorganism. Archival FFPE tissues were examined from ten mice, and five nonhuman primates after exposure to B. pseudomallei or B. mallei by LSCM. Additionally, a historical spleen biopsy from a human suspected of exposure to B. mallei was examined. B. pseudomallei was seen in many of the infected tissues from mice. Four out of five nonhuman primates were positive for the pathogen. In the human sample, B. mallei was seen in pyogranulomas in the spleen biopsy. Thus, the presence of the pathogen was validated by LSCM in murine, nonhuman primate, and human FFPE tissues.
关键词： melioidosis,Burkholderia pseudomallei,Burkholderia mallei,animal models,microorganism,formalin-fixed paraffin embedded tissue,laser scanning confocal microscopy,glanders
更新于2025-09-23 15:21:01
-
Calibration of a Chromatic Confocal Microscope for Measuring a Colored Specimen
摘要： In this paper, a color correction method is proposed to improve measurement accuracy for chromatic confocal microscopy (CCM) when measuring a colored specimen. Characteristic curve shifting due to selective re?ection from color surfaces was analyzed based on a laboratory CCM system developed by the authors’ team. Theoretically, when the color of the targeted surface is different from that represented by the central wavelength of the light source, the characteristic curve of CCM would have a notable deviation from that of an achromatic surface. In this study, this conclusion was veri?ed through both simulation and experiments. Using a set of standard color calibration pieces, a color correction method was proposed accordingly to quantify the characteristic curve shifting. To validate the proposed method, a laser scanning confocal microscope Carl Zeiss LSM700 was used as the referencing system. Experimental data showed that with the color correction method, measurement errors can be controlled within 10 nm. Compared with the measurement without color correction, the measurement accuracy is signi?cantly improved.
关键词： characteristic curve shifting,Chromatic confocal microscopy,colour correction curve.,colour specimen,spectral information
更新于2025-09-23 15:21:01