Background and aims: Probe-based confocal laser endomicroscopy (pCLE) is used to differentiate between neoplastic and non-neoplastic colorectal polyps during colonoscopy. We aimed to assess the accuracy of two endoscopists starting to use real-time pCLE for differentiation of colorectal polyps and to determine the negative predictive value (NPV) for neoplasia in ... read more polyps ≤ 5 mm. Methods: Patients undergoing colonoscopy in a tertiary hospital were included in this prospective trial. After a training session, two colonoscopists assessed 50 polyps between August 2012 and April 2014. They sequentially used narrow-band imaging (NBI) and real-time pCLE to differentiate non-adenomatous, adenomatous, and carcinomatous polyps during colonoscopy. Histologic diagnosis by a gastrointestinal pathologist was the gold standard. Results were compared to post-hoc pCLE by a panel of gastroenterologists and pathologists. Results: The accuracy of real-time pCLE was 76 %, compared to 73 % for NBI, and ...
TY - JOUR. T1 - Probe-based confocal laser endomicroscopy (pCLE) images of submucosal growth of a duodenal mucous neck cell adenoma. AU - Tahara, Tomomitsu. AU - Horiguchi, Noriyuki. AU - Nagasaka, Mitsuo. AU - Nakagawa, Yoshihito. AU - Tsukamoto, Tetsuya. AU - Shibata, Tomoyuki. AU - Ohmiya, Naoki. PY - 2016/1/1. Y1 - 2016/1/1. UR - http://www.scopus.com/inward/record.url?scp=84955452020&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=84955452020&partnerID=8YFLogxK. U2 - 10.1055/s-0041-111031. DO - 10.1055/s-0041-111031. M3 - Comment/debate. C2 - 26800196. AN - SCOPUS:84955452020. VL - 48. SP - E19-E21. JO - Endoscopy. JF - Endoscopy. SN - 0013-726X. ER - ...
TY - JOUR. T1 - Pre- and post-training session evaluation for interobserver agreement and diagnostic accuracy of probe-based confocal laser endomicroscopy for biliary strictures. AU - Talreja, Jayant P.. AU - Turner, Brian G.. AU - Gress, Frank G.. AU - Ho, Sammy. AU - Sarkaria, Savreet. AU - Paddu, Naveen. AU - Natov, Nikola. AU - Bharmal, Sheila. AU - Gaidhane, Monica. AU - Sethi, Amrita. AU - Kahaleh, Michel. PY - 2014/7. Y1 - 2014/7. N2 - Background and Aim Current diagnostic modalities for indeterminate biliary strictures offer low accuracy. Probe-based confocal laser endomicroscopy (pCLE) permits microscopic assessment of mucosal structures by obtaining real-time high-resolution images of the mucosal layers of the gastrointestinal tract. Previously, an interobserver study demonstrated poor to fair agreement even among experienced confocal endomicroscopy operators. Our objective was to assess interobserver agreement and diagnostic accuracy upon completion of a pCLE training session. Methods ...
New approaches to the chemotherapy of glioblastoma [Elektronische Ressource] : investigations on doxorubicin nanoparticles, inhibition of PDGF receptors and kinesin Eg5, with emphasis on confocal laser-scanning microscopy / vorgelegt von Dietmar Gross : New Approaches to the Chemotherapy of Glioblastoma: investigations on doxorubicin nanoparticles, inhibition of PDGF receptors and kinesin Eg5, with emphasis on confocal laser-scanning microscopy Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. nat.) der Naturwissenschaftlichen Fakultät IV - Chemie und Pharmazie - der Universität Regensburg
TY - JOUR. T1 - In vivo reflectance confocal microscopy for varicella prompt diagnosis and treatment in a severely immunosuppressed patient. AU - Abraham, Leonardo Spagnol. AU - Costa, Mariana Carvalho. AU - Agozzino, Marina. AU - Amorosi, Beatrice. AU - Cota, Carlo. AU - Ardigo, Marco. PY - 2012/8. Y1 - 2012/8. UR - http://www.scopus.com/inward/record.url?scp=84863550245&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=84863550245&partnerID=8YFLogxK. U2 - 10.1111/j.1600-0846.2011.00581.x. DO - 10.1111/j.1600-0846.2011.00581.x. M3 - Article. C2 - 22092934. AN - SCOPUS:84863550245. VL - 18. SP - 386. EP - 388. JO - Skin Research and Technology. JF - Skin Research and Technology. SN - 0909-752X. IS - 3. ER - ...
A Wavelet-Based Approach to 3-D Confocal Microscopy Image Reconstruction , A Wavelet-Based Approach to 3-D Confocal Microscopy Image Reconstruction , کتابخانه دیجیتال دانشگاه آزاد اسلامی خوراسگان
TY - JOUR. T1 - Interobserver agreement of confocal laser endomicroscopy for detection of head and neck neoplasia. AU - Moore, Charles. AU - Mehta, Vikas. AU - Ma, Xiaohui. AU - Chaudhery, Shabnum. AU - Shi, Runhua. AU - Moore-Medlin, Tara. AU - Lian, Timothy. AU - Nathan, Cherie Ann O.. PY - 2016/3/1. Y1 - 2016/3/1. N2 - Objectives/Hypothesis We have described the feasibility of using the probe-based confocal laser endomicroscopy (pCLE) in differentiating benign from malignant lesions of the head and neck. Therefore, we wanted to determine the interobserver agreement of pCLE offline images of noncancerous, precancerous, and cancerous lesions of the head and neck. Study Design Single tertiary referral center. Methods In the feasibility study, image criteria for nondysplasia, dysplasia, and cancer were defined. The pCLE was performed before lesions were biopsied. Fifty offline images and 10 videos of good quality were selected. Seven surgeons and one pathologist were asked to review and ...
A novel, noninvasive imaging technique for intraoperative assessment of parathyroid glands: confocal reflectance microscopy. Surgery. 2000 Dec; 128(6):1088-1100; discussion 1100-1 ...
INTRODUCTION: Endoscopic evaluation with high-definition white light endoscopy and random 4-quadrant biopsy (Seattle Protocol) is the current standard of care for the detection of Barretts esophagus (BE). Recently, enhanced imaging technologies have become available to provide real-time diagnosis of intestinal metaplasia (IM) and dysplasia, reducing the need for tissue biopsy. Probe-based confocal laser endomicroscopy (pCLE) provides dynamic microscopic mucosal views, rapidly capturing digital images that become optical biopsies. This study examined the role of pCLE in BE screening and surveillance as compared to the Seattle Protocol.. METHODS: Patients undergoing BE screening or surveillance endoscopy were enrolled at eight US centers. Optical biopsy using pCLE was interpreted in real time. Endoscopists performing pCLE were new users with a median experience of 8.5 months and no formal training in surgical pathology. Seattle Protocol biopsies were then taken. Recorded pCLE images were reviewed ...
TY - JOUR. T1 - First experiences using reflectance confocal microscopy on equivocal skin lesions in Queensland. AU - Curchin, Claudia E S. AU - Wurm, Elisabeth M T. AU - Lambie, Duncan L J. AU - Longo, Caterina. AU - Pellacani, Giovanni. AU - Soyer, H. Peter. PY - 2011/5. Y1 - 2011/5. N2 - Background/Objectives: Reflectance confocal microscopy (RCM) is a non-invasive method of imaging human skin in vivo. The purpose of this study was to observe the experience of using RCM on equivocal skin lesions in a tertiary clinical setting in Queensland. Methods: Fifty equivocal lesions on 42 patients were imaged using a reflectance confocal microscope immediately prior to being excised. The images were then analysed blind to the histopathological diagnosis. The experience and problems encountered when using RCM on skin lesions for the first time was also observed. Results: On RCM analysis 12/13 melanomas (92.3% sensitivity, 75% specificity), 19/22 benign naevi (86% sensitivity, 95% specificity), 6/9 basal ...
Representative confocal microscopy images of cells exposed to green fluorescent nano-particles (40 nm in diameter). (A-C) Dural epithelial cells. (D-F) Dura
A prospective randomized study of 26 patients treated with flex in one eye, and smile in the other at the Department of Ophthalmology, Aarhus University Hospital. Preoperative spherical equivalent refraction averaged -7.50 D (range -6.00 to -9.75 D) in both groups. All patients had stable myopia and no other ocular diseases. A VisuMax® femtosecond laser (Carl Zeiss Meditec, Jena, Germany) was used. Lenticule diameters were the same in both eyes and ranged from 6.00 to 6.50 mm. Flap thickness was 110 to 120 ṁm, and flap/cap diameter ranged from 7.3 to 8.0 mm. Images of the subbasal corneal nerves were acquired by continuous through-focusing of the cornea using in vivo confocal laser-scanning microscopy (Heidelberg Engineering, Heidelberg, Germany). The detachable objective, the so called Rostock cornea module (RCM) , was used to optimize image acquisition. The NeuronJ Java program (http://www.imagescience.org/meijering/software/neuronj) was used to trace and quantify subbasal nerves. Mean ...
The pUL97 protein kinase encoded by human cytomegalovirus is a multifunctional determinant of the efficiency of viral replication and phosphorylates viral as well as cellular substrate proteins. Here, we report that pUL97 is expressed in two isoforms with molecular masses of approximately 90 and 100 kDa. ORF UL97 comprises an unusual coding strategy in that five in-frame ATG start codons are contained within the N-terminal 157 aa. Site-directed mutagenesis, transient expression of point and deletion mutants and proteomic analyses accumulated evidence that the formation of the large and small isoforms result from alternative initiation of translation, with the start points being at amino acids 1 and 74, respectively. In vitro kinase assays demonstrated that catalytic activity, in terms of autophosphorylation and histone substrate phosphorylation, was indistinguishable for the two isoforms. An analysis of the intracellular distribution of pUL97 by confocal laser-scanning microscopy demonstrated that both
Glomeruli are anatomical and possibly functional modules in the vertebrate olfactory bulb. We investigated the spatial arrangement of glomeruli in the olfactory bulbs of adult zebrafish (Brachydanio rerio). A solution of the lipophilic tracer Dil was injected into the nasal cavities. Axons of sensory neurons projecting from the olfactory epithelium into the bulb were traced anterogradely, thus labeling the whole population of glomeruli. The glomerular distribution was analyzed in detail by confocal laser-scanning microscopy. We find that a typical olfactory bulb contains a small number of about 80 glomeruli that have a stereotyped configuration in all animals investigated. All glomeruli exhibit bilateral symmetry. Twenty-two single glomeruli could be identified from animal to animal by their characteristic position and morphology. The remaining glomeruli either are embedded in glomerular plexus and therefore cannot be delineated reliably, or belong to a densely clustered subpopulation of on ...
We present a low-cost, high-speed, retrofitted laser scanning module for microscopy. The cage-mounted system, with various available fiber-coupled sources, offers a real-time imaging alternative to costly commercial systems with capabilities for conventional or confocal reflectance and fluorescence applications as well as advanced laser scanning microscopy implementations. Reflectance images of a resolution target and confocal images of fluorescent polystyrene beads are presented for system characterization. Confocal fluorescence image stacks of T84 epithelial cancer cells are presented to demonstrate application to biological studies. This laser scanning module is a flexible, scalable, high-speed alternative to commercial laser scanning systems suitable for applications requiring a simple imaging tool and for teaching laboratories.. © 2005 Optical Society of America. Full Article , PDF Article ...
Cell culture and toxicity studies. Human embryonic kidney cells (HEK293) were cultivated in DMEM supplemented with 10% FCS and 2 mM L-glutamine at 37°C and 5% CO2. Primary neuron cultures were prepared from hippocampi or cortex of C57BL/6 mice and cultured in neurobasal medium supplemented with B-27, N-2, and L-glutamine. All reagents were supplied by Life Technologies (Thermo Fisher Scientific). Neurons were cultivated at different densities depending on which culture plate was used (30,000/well for 96-well plates, 75,000/well for 24-well plates, and 900,000/well for 6-well plates). For confocal laser-scanning microscopy, neurons and HEK293 cells were cultivated on poly-L-lysine-coated and collagenized coverslips, respectively. Unless otherwise stated, all neuronal studies were conducted after 9 to 10 days in vitro (DIV).. Toxicity studies using the MTT assay were conducted in 96-well plates for 12 hours in the presence of 200 μM of each investigated metabolite, unless otherwise stated. For ...
Confocal laser-scanning microscopy was used to carry out a comparative study of the immunostaining for three families of neuropeptides, viz., allatostatin-A (AS-A), allatostatin-C (AS-C) and allatotropin (AT), in adult female mosquitoes of Aedes aegypti and Anopheles albimanus. The specific patterns of immunostaining for each of the three peptides were similar in both species. The antisera raised against AT, AS-A, and AS-C revealed intense immunoreactivity in the cells of each protocerebral lobe of the brain and stained cells in each of the ventral ganglia and neuronal projections innervating various thoracic and abdominal tissues. Only the AS-A antiserum labeled immunoreactive endocrine cells in the midgut. The distribution of the peptides supports the concept that they play multiple regulatory roles in both species.
To quantify the degree of alignment in the data sets, the authors use a grayscale moments analysis of the data to obtain a histogram of the orientations of the fibers in the sample. This yields values of the azimuthal angle ,math,\phi,/math, defined within the imaging plane and of the polar angle ,math,\theta,/math, defined with respect to the perpendicular (z) axis. Since the area of a surface element for a unit sphere is ,math,sin \theta d\theta d\phi,/math,, an isotropic network should show a sine distribution for ,math,\theta,/math, and a uniform distribution for ,math,\phi,/math,. The distributions are shown in Figure 2. The ,math,\phi,/math, distribution is fairly flat for both imaging methods, revealing that the sample is isotropic within the focal plane. For ,math,\theta,/math,, CFM data follows a sine distribution whereas CRM data deviates strongly. To test whether the anisotropy in the CRM data is an imaging artifact, the authors rotate the sample by 90 degrees. Whereas the CFM data ...
The enamel defects (EDs) may present with a variety of clinical manifestations with increasing severity from the sole appearance of pale discoloration to remarkable structural alterations. EDs are responsible for higher caries receptivity. In vivo reflectance confocal microscopy (RCM) allows to image in vivo at microscopic resolution of the dental surface, thus avoiding the tooth extraction and the sample preparation because of its ability to optically scan living tissues along their depth. Aim of this study is the in vivo assessment at microscopic resolution of dental surfaces affected by EDs without resorting to invasive methods such as teeth extractions, to define histological findings occurring in chromatic and/or structural EDs. For the purpose, 15 children, referring at the Dental Clinic of the Second University of Naples, affected by several degrees of EDs, were enrolled and underwent in vivo RCM imaging to microscopically define the ED confocal features using a commercially available ...
Buchner AM, Gomez V, Heckman MG, Shahid MW, Achem S, Gill KR, Jamil LH, Kahaleh M, Lo SK, Picco M, Riegert-Johnson D, Raimondo M, Sciemeca D, Wolfsen H, Woodward T, Wallace MB. The learning curve of in vivo probe-based confocal laser endomicroscopy for prediction of colorectal neoplasia. Gastrointest Endosc. 2011 Mar; 73(3):556-60 ...
Plants are valuable systems for analyzing the acentriolar mitotic spindle. We have developed methods for imaging the mitotic spindle in living tobacco (Nicotiana tabacum) suspension culture cells expressing GFP-?-tubulin. The methods allow the spindle to be observed in living cells at high spatial and temporal resolution and rely on water immersion objectives, spinning disk optics, and high-sensitivity cameras. Here, we describe these methods and provide step-by-step protocols for certain key steps. We also describe a method for application and removal of inhibitors ...
Breast cancer is the most prevalent and deadly cancer among women worldwide. The current standard for breast lesion diagnosis is histologic assessment with hematoxylin and eosin (H&E) staining. Histology has high diagnostic accuracy, but requires extensive time and resources to perform. The objective of this work was to improve diagnosis of early breast cancers by developing approaches to rapidly image and characterize neoplastic tissue and the tumor microenvironment in high resolution optical images. Confocal fluorescence microscopy can image optical sections of tissue without the need for extensive tissue processing. Three studies were performed to evaluate if confocal microscopy images contain sufficient information to identify neoplasia in breast tissue. In a 31 patient study, five pathologists identified neoplasia with high accuracy in confocal and histologic images. In another study, an expert pathologist estimated tumor cellularity in core biopsies with moderate agreement between confocal ...
The use of blochemical fractionation, immunofluorescence laser-scanning confocal microscopy, and immunoelectron microscopy with mouse anti-human bcl-2 monoclonal antibody to analyze the subcellular localization of the bcl-2 gene product revealed the protein prominently in the nuclear envelope, endoplasmic reticulum membrane, and mitochondrial membranes. Electron microscopy at high magnification more precisely localized bcl-2 to the nuclear outer membrane as confirmed by the biochemical fractionation, as well as to mitochondrial outer and, to a lesser degree, inner membrane. This multisite membrane distribution of bcl-2 suggests an important role for this protein in several different membrane compartments.. ...
When T cells, B cells, and natural killer (NK) cells of the immune system interact with target cells, signaling molecules are accumulated in the plasma membrane at structures known as the immunological synapse. Evidence is accumulating that proteins, as well as signals, are transferred between the interacting cells at such contacts. NK cells receive inhibitory signals from cells that express self major histocompatibility complex (MHC) molecules on their surface. Earlier evidence had shown that NK cells can actually acquire MHC class I proteins during interactions with target cells. Now Vanherberghen et al. show that the exchange goes both ways and that NK receptors are transferred to cells that express MHC class I ligands. The authors monitored transfer of biotinylated killer Ig-like receptor (KIR) KIR2DL1 by immunoblotting or green fluorescent protein-tagged receptor by fluorescence-activated cell sorting or laser-scanning confocal microscopy and observed transfer of KIRs. The NK cell receptor ...
Areas of active investigation include: use of laser-scanning confocal microscopy to measure calcium sparks, which are brief localized increases in fluorescence from a Ca-indicator such as fluo-3 that are thought to be reflective of the transient opening of one or a few RyRs (=ryanodine receptors), the Ca release channels of the sarcoplasmic reticulum (SR); the possibility that the mechanism of activation of RyRs involves both voltage-gating and Ca-gating; the nature of the mechanism whereby SR Ca release is inactivated by a rise in myoplasmic free [Ca]; the possibility that either activation or inactivation of SR Ca release may vary with the RyR isoform composition (RyR1, RyR3, etc.); estimation of local Ca movements within the sarcomere by means of computer modeling, including estimation of the kinetics of binding of Ca to the intracellular Ca buffers troponin, parvalbumin, ATP, and the SR Ca pump ...
Plant parasites secrete proteins known as effectors into host tissues to manipulate host cell structures and functions. One of the major goals in effector biology is to determine the host cell compartments and the protein complexes in which effectors accumulate. Here, we describe a five-step pipeline that we routinely use in our lab to achieve this goal, which consists of (1) Golden Gate assembly of pathogen effector-green fluorescent protein (GFP) fusions into binary vectors, (2) Agrobacterium-mediated heterologous protein expression in Nicotiana benthamiana leaf cells, (3) laser-scanning confocal microscopy assay, (4) anti-GFP coimmunoprecipitation-liquid chromatography-tandem mass spectrometry (coIP/MS) assay, and (5) anti-GFP western blotting ...
Cells were fixed with 3% PFA in warm PHEM buffer (60 mM Pipes-KOH, pH 6.9, 25 mM Hepes, 10 mM EDTA, and 2 mM MgCl2) containing 0.1% Triton X-100. EB1 staining required fixation with methanol at −20°C for 5 min. Primary antibodies to EB1 (1A11/4; Santa Cruz Biotechnology, Inc.), paxillin (349; BD), SEPT2 (N5N; gift from M. Kinoshita, Nagoya University, Nagoya, Japan), SEPT6 (S6CU; gift from M. Kinoshita), SEPT7 (Immuno-Biological Laboratories, Inc.), GFP (Invitrogen), α-tubulin (DM1A; Sigma-Aldrich), and secondary donkey DyLight 488-, 549-, 594-, or 649-conjugated F(ab′)2 to mouse or rabbit IgG (Jackson ImmunoResearch Laboratories, Inc.) were diluted in PBS with 2% BSA. Staining with antibodies of the same species was performed with an antibody-labeling kit (Zenon; Invitrogen). Coverslips were mounted with FluorSave (EMD) or Vectashield (Vector Laboratories) and imaged on a confocal laser-scanning microscope (FluoView 1000; Olympus) using a Plan Apochromat 60×/1.42 NA objective. ...
Fluorescence confocal laser scanning endomicroscopy is a novel tool combining confocal microscopy and endoscopy for in-vivo subcellular structure imaging with comparable resolution as the traditional microscope. In this paper, we propose a three-channel fluorescence confocal microscopy system based on fiber bundle and two excitation laser lines of 488nm and 650nm. Three fluorescent photomultiplier detecting channels of red, green and blue can record multi-color fluorescence signals from single sample site simultaneously. And its ability for in-vivo multi-channel fluorescence detection at subcellular level is verified. Moreover, the system has achieved an effective field of view of 154μm in diameter with high resolution. With its multi-laser scanning, multi-channel detection, flexible probing, and in-vivo imaging abilities it will become a powerful tool in bio-chemical research and diagnostics, such as the investigation of the transport mechanism of nano-drugs in small animals.. ...
By enhancing the signal of genetically encoded markers expressed in defined circuit elements and quickly mapping them in large volumes of brain tissue, it enables sparse reconstructions of genetically defined circuit motifs. These motifs can be further used as road maps for targeted imaging of tissue subsets at high resolution, thus restricting imaging and segmentation time by enabling directed unsupervised reconstructions. As a proof of principle, this technique has been used to automatically reconstruct and visualize interneurons of different mouse retinas. This approach is not restricted to the retina and can be used to track long-range projections anywhere in the brain, e.g., from the retina to visual recipient layers located several millimeters away. Further, the same approach for unsupervised image processing can be applied to a variant of spectral confocal reflectance microscopy, facilitating the long range tracing of myelinated axons, as well as the automatic assessment of myelin ...
A variety of microscopic techniques have been utilized to study cyanobacterial associations with plant roots, but confocal laser scanning microscopy (CLSM) is the least used due to the unavailability of a suitable fluorescent dye. Commonly used lectins have problems with their binding ability with root cells and their visualization under CLSM. DTAF (5-(4,6-dichlorotriazinyl) aminofluorescein) is a fluorescent dye that has been widely used for staining various biological samples for fluorescent microscopy. It reacts with polysaccharides and peptides at ordinary conditions. The possible application and efficiency of DTAF for CLSM studies were examined in various aspects of cyanobacterial-plant interactions. Seedlings of Pisum sativum, Vigna rediata and Triticum aestivum were co-cultivated and stained with DTAF as a fluorochrome. Extracellular and intracellular interactions of cyanobacteria and the plant root surface were observed by CLSM. Results were compared with staining by other commonly used ...
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For these reasons, in vivo observation of the skin in its physiology and responses to stimuli has always been somewhat of a challenge. What Corcuff, P et al experimented with was a method of observing the skin of the forearm before sun exposure and for a month afterward using confocal microscopy techniques. Observations were made in regards to melanasome (melanin-containing organelle) caps within basal keratinocytes (skin cells in the deep epidermis). The use of in vivo confocal microscopy also affords new insight to the role of melanin and its gradual migration after sun exposure In vivo confocal microscopy involved reflected singals being used to create an image. Contrast was improved due to absorption and scattering (minimal loss of photons). Another feature that was regarded was realtime imaging to prevent blurring from motion caused by blood pulses and involuntary movement. ...
Multi-dimensional fluorescence imaging of live animal models demands strong optical sectioning, high spatial resolution, fast image acquisition, and minimal photobleaching. While conventional laser scanning microscopes are capable of deep penetration and sub-cellular resolution, they are generally too slow and causing excessive photobleaching for volumetric or time-lapse imaging. We demonstrate the performance of an augmented line-scan focal modulation microscope (aLSFMM), a high-speed imaging platform that affords above video-rate imaging speed by the use of line scanning. Exceptional background rejection is accomplished by combining a confocal slit with focal modulation. The image quality is further improved by merging the information from simultaneously acquired focal modulation and confocal images. Such a hybrid imaging scheme makes it possible to use very low power excitation light in high-speed imaging, and therefore leads to reduced photobleaching that is desirable for three-dimensional ...
Intra-cellular and inter-cellular protein translocation can be observed by microscopic imaging of tissue sections prepared immunohistochemically. A manual densitometric analysis is time-consuming, subjective and error-prone. An automated quantification is faster, more reproducible, and should yield results comparable to manual evaluation. The automated method presented here was developed on rat liver tissue sections to study the translocation of bile salt transport proteins in hepatocytes. For validation, the cholestatic liver state was compared to the normal biological state. An automated quantification method was developed to analyze the translocation of membrane proteins and evaluated in comparison to an established manual method. Firstly, regions of interest (membrane fragments) are identified in confocal microscopy images. Further, densitometric intensity profiles are extracted orthogonally to membrane fragments, following the direction from the plasma membrane to cytoplasm. Finally, several
In this issue, Wen and colleagues demonstrate that inhibition of the prolactin receptor by G129R initiates FOXO3a-mediated cell death in ovarian cancer. The confocal microscopy image on this months cover demonstrates treatment with G129 results in nuclear accumulation of FOXO3a (in red) in a uterine cancer cell line. They further demonstrate FOX3a and EIF-4EBP1 play a critical role in the cytotoxic response to G129R. Read the full article on page 1943. ...
Synthesis of a 1:1 mixture of lactose/mannose-QDs 20 b and lactose/maltotriose-QDs 21 b and confocal microscopy images after 2 h incubation with AS cells: A)
We established a collection of 7,000 transgenic lines of Drosophila melanogaster. Expression of GAL4 in each line is controlled by a different, defined fragment of genomic DNA that serves as a transcriptional enhancer. We used confocal microscopy of dissected nervous systems to determine the expression patterns driven by each fragment in the adult brain and ventral nerve cord. We present image data on 6,650 lines. Using both manual and machine-assisted annotation, we describe the expression patterns in the most useful lines. We illustrate the utility of these data for identifying novel neuronal cell types, revealing brain asymmetry, and describing the nature and extent of neuronal shape stereotypy. The GAL4 lines allow expression of exogenous genes in distinct, small subsets of the adult nervous system. The set of DNA fragments, each driving a documented expression pattern, will facilitate the generation of additional constructs for manipulating neuronal function. synapse was substantially ...
Opterra II swept field scanning confocal microscope Brochure 2.8 MB Opterra II is the latest advancement in high-speed fluorescence microscopy designed specifically for live-cell microscopy. It uniquely combines the resolution of traditional confocal systems with the speed typically associated with wide-field imaging. ...
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The FCCM Facility was developed to assist investigators with the use of several different sophisticated instruments designed to detect and measure fluorescent light emission. A BD FACSCalibur flow cytometer is available for analysis of cells and particles from each other based on size, internal complexity, and/or up to four different fluorescent signals. Cells labeled either internally or externally with fluorescent antibodies, calcium or pH specific probes, fluorescent proteins, or with DNA specific probes and dyes. Cells or particles can also be aseptically sorted to obtain pure populations for either further analysis or subsequent culture. A Leica SP2 laser scanning confocal microscope and an automated Zeiss Axiovert 200M driven by OpenLab software are available for imaging of live or fixed cells or other fluorescent materials. A spinning disk confocal and TIRF microscopy system will be added to the facility in 2008.
The Opterra II swept-field confocal microscope utilizes proprietary one-dimensional pinhole array technology to combine the resolution of traditional confocal systems with the speed typically associated with wide-field imaging. With its short acquisition times and cell-protecting minimization of photobleaching and phototoxicity, Opterra II is ideal for advanced live-cell imaging. ...
The first four members of the myeloid differentiation marker 88 (MyD88) family of cytosolic adaptor proteins are well known for their roles in mediating Toll-like receptor (TLR) signaling in the immune system. Kim et al. set out to characterize the more enigmatic fifth member of this family, MyD88-5. Northern and Western blotting demonstrated a higher abundance of MyD88-5 mRNA and protein in the brains of mice and humans than in any other organ. In human blood, real-time reverse transcription polymerase chain reaction (RT-PCR) assays showed that MyD88-5 mRNA was preferentially found in lymphocytes and not in myeloid cells. The authors developed a transgenic mouse that expressed a green fluorescent protein (GFP)-tagged MyD88-5 protein. Confocal microscopic analysis of brain sections showed that GFP fluorescence was strongest in the hippocampus and cerebellum and found only in neurons, within which MyD88-5-GFP was cytoplasmic and had a punctate distribution. When expressed in COS-1 cells, ...
HCS systems are basically fully automated microscopes: an automated stage moves the multi-well plate from position to position (according to a previously defined protocol) and at each position the same imaging procedure is performed. Dichroic mirrors and filters are automatically switched, the sample is illuminated at one or more excitation wavelengths and the images are recorded with a CCD camera. Optionally also brightfield images are recorded. A combination of a laser-based autofocus system and a software-based autofocus procedure ensure that the sample is always in focus during imaging. HCS systems are available in widefield configuration or as confocal setup as for certain experiments optical sections or 3D imaging is required. The confocal HCS systems usually employ a spinning disk technique to ensure fast image acquisition. For fluorescence excitation the systems are equipped either with Mercury or Xenon light sources or with lasers, which are frequently used in confocal HCS systems. When ...
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One of the key frontiers in optical imaging is to maximize the spatial information retrieved from a sample while minimizing acquisition time. Confocal laser scanning microscopy is a powerful imaging modality that allows real-time and high-resolution acquisition of two-dimensional (2D) sections. However, in order to obtain information from threedimensional (3D) volumes it is currently limited by a stepwise process that consists of acquiring multiple 2D sections from different focal planes by slow z-focus translation. Here, we present a novel method that enables the capture of an entire 3D sample in a single step. Our approach is based on an acoustically-driven varifocal lens integrated in a commercial confocal system that enables axial focus scanning at speeds of 140 kHz or above. Such high-speed allows for one or multiple focus sweeps on a pixel by pixel basis. By using a fast acquisition card, we can assign the photons detected at each pixel to their corresponding focal plane allowing ...
In the MBS program, Barthel, who didnt take many molecular courses as an undergraduate, enrolled in a variety of molecular and cell biology, spectroscopy, cell physiology, and other courses.. The incredible access to University faculty and resources helped him get a paper published with Karen Mesce and several others about a new confocal microscopy technique used to image Golgi-stained neurons.. It seems like everyone you work with around here is a genius. My knowledge base now has far surpassed what it was.. He also found a full-time job on campus at the Imaging Center, where he provided training, as well as imaged specimens such as intact and optically clear mouse brains in order to study viral induced genetic modification. (Imaging Center program director, Mark Sanders, was also a co-author of the paper and instrumental in Barthels success there.). They have a lot of impressive high-end research equipment that they teach people how to use, in addition to providing sample preparation, ...
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Using the 3D spinning disk confocal microscope at CBST, we are able to observe this process in much greater detail (more than 10 times the useful information in terms of spatial and temporal resolution), and reconstruct video images to focus on the structure and behavior of the virological synapse, which forms at the interface of cells in contact. By uncovering the different modes of Gag movements in the donor cell, we may be able to identify new targets or strategies for antiviral therapy.. ...
Magnified intelligence chromoendoscopy (FICE) plus probe-based confocal laser endomicroscopy (pCLE) for Gastric Intestinal Metaplasia (GIM) diagnosis: a feasibility trial. Research Question:. Is confocal endomicroscope feasible to diagnose gastric intestinal metaplasia?. Objective:. To evaluate the feasibility of confocal endomicroscope in diagnose gastric intestinal metaplasia.. Hypothesis:. Confocal endomicreosocpe can provide the accurate diagnosis of gastric intestinal metaplasia.. Research design:. Diagnostic study. Sample size:. The investigators follow the population in recent study from Imraporn et al.: Validity of magnify NBI for gastric intestinal metaplasia targeted biopsy (N= 50). Data analysis:. Confocal Barretts esophagus classification was used to evaluate agreement of confocal endomicroscopic finding in gastric intestinal metaplasia. The accuracy of new criteria for GIM by confocal endomicroscope was evaluated in relation to pathological report, a gold standard for diagnosis, ...
TY - CONF. T1 - Semi-adaptive optics in confocal reflection and two photon microscopy in rat brain. AU - Vijverberg, J.. AU - Poland, S.P.. AU - Wright, A.. AU - Girkin, J.M.. PY - 2007. Y1 - 2007. N2 - Paper concerned with semi-adaptive optics in confocal reflection and two photon microscopy in rat brain.. AB - Paper concerned with semi-adaptive optics in confocal reflection and two photon microscopy in rat brain.. KW - semi-adaptive optics. KW - confocal reflection. KW - photon microscopy. KW - rat brain. KW - microscopy. M3 - Paper. T2 - Photon 06. Y2 - 4 September 2006 through 7 September 2006. ER - ...
This study aims to investigate the roles of the Notch-Hes1 pathway in the advanced glycation end product (AGE)-mediated differentiation of neural stem cells (NSCs). We prepared pLentiLox3.7 lentiviral vectors that express short hairpin RNA (shRNA) against Notch1 and transfected it into NSCs. Cell differentiation was analyzed under confocal laser-scanning microscopy. The percentage of neurons and astrocytes was quantified by normalizing the total number of TUJ1+ (Neuron-specific class III β-tubulin) and GFAP+ (Glial fibrillary acidic protein) cells to the total number of Hoechst 33342-labeled cell nuclei. The protein and gene expression of Notch-Hes1 pathway components was examined via western blot analysis and real-time PCR. After 1 week of incubation, we found that AGE-bovine serum albumin (BSA) (400 μg/mL) induced the astrocytic differentiation of cultured neurospheres and inhibited neuronal formation. The expression of Notch-Hes1 pathway components was upregulated in the cells in the AGE-BSA
TY - JOUR. T1 - Two-photon laser confocal microscopy of micropermeability of resin-dentin bonds made with water or ethanol wet bonding. AU - Sauro, Salvatore. AU - Watson, Timothy F.. AU - Mannocci, Francesco. AU - Miyake, Katsuya. AU - Huffman, Bradford P.. AU - Tay, Franklin R.. AU - Pashley, David H.. N1 - Copyright: Copyright 2015 Elsevier B.V., All rights reserved.. PY - 2009/7. Y1 - 2009/7. N2 - This study evaluated the micropermeability of six etch-and-rinse adhesives bonded to dentin. There were two principal groups: wet bonding with water or wet bonding with absolute ethyl alcohol. After bonding and the creation of composite build-ups, the pulp chambers were filled with 0.1% lucifer yellow. The contents of the pulp chamber were kept under 20 cm H2O pressure to simulate pulpal pressure for 3 h. The specimens were vertically sectioned into multiple 0.5-mm thick slabs that were polished and then examined using a twophoton confocal laser scanning microscope (TPCLSM). The results showed that ...
Clinical applications of corneal confocal microscopy Mitra Tavakoli1, Parwez Hossain2, Rayaz A Malik11Division of Cardiovascular Medicine, University of Manchester and Manchester Royal Infirmary, Manchester, UK; 2University of Southampton, Southampton Eye Unit, Southampton General Hospital, Southampton, UKAbstract: Corneal confocal microscopy is a novel clinical technique for the study of corneal cellular structure. It provides images which are comparable to in-vitro histochemical techniques delineating corneal epithelium, Bowman’s layer, stroma, Descemet’s membrane and the corneal endothelium. Because, corneal confocal microscopy is a non invasive technique for in vivo imaging of the living cornea it has huge clinical potential to investigate numerous corneal diseases. Thus far it has been used in the detection and management of pathologic and infectious conditions, corneal dystrophies and ecstasies, monitoring contact lens induced corneal changes and for pre and post surgical evaluation
Confocal laser endomicroscopy (CLE) is a new diagnostic technique that allows microscopic examination of the digestive mucosa during ongoing endoscopy. Different types of tissue and diseases can be diagnosed immediately, and analysis of the in vivo microarchitecture is helpful to better target standard biopsies and reduce the number of biopsies required. CLE necessitates an intravenous injection of a fluorescent marker, e.g. fluorescein, to obtain optical biopsies with a high level of magnification (up to 1000 fold). To date, more than 1000 endomicroscopy procedures have been performed in the world and different publications have shown the safety, feasibility and excellent diagnostic yield of CLE. No complication related to IV injection of fluorescein has been reported. However, all these data come from a very limited number of expert centres and need to be confirmed and validated at the multicenter level. The aims of this multicenter trial are: 1) to standardize CLE in all centres equipped in ...
TY - JOUR. T1 - A non-contact measuring system for in-situ surface characterization based on laser confocal microscopy. AU - Fu, Shaowei. AU - Cheng, Fang. AU - Tjahjowidodo, Tegoeh. AU - Yu, Zhou. AU - Butler, David. PY - 2018/8/13. Y1 - 2018/8/13. N2 - The characterization of surface topographic features on a component is typically quantified using two-dimensional roughness descriptors which are captured by off-line desktop instruments. Ideally any measurement system should be integrated into the manufacturing process to provide in-situ measurement and real-time feedback. A non-contact in-situ surface topography measuring system is proposed in this paper. The proposed system utilizes a laser confocal sensor in both lateral and vertical scanning modes to measure the height of the target features. The roughness parameters are calculated in the developed data processing software according to ISO 4287. To reduce the inherent disadvantage of confocal microscopy, e.g. scattering noise at steep ...
Random biopsies with targeted biopsies of any suspicious lesions is a reasonable alternative if chromoendoscopy is not available or if the yield of chromoendoscopy is reduced by significant underlying inflammation, pseudopolyposis, or poor preparation (ASGE, 2015). While there is emerging evidence that chromoendoscopy may yield higher polyp detection rates, at this time it is not known if the additional polyps detected are clinically significant and whether a higher detection rate results in a meaningful clinical outcome benefit. Confocal Laser Endomicroscopy Confocal laser endomicroscopy (also known as confocal fluorescent endomicroscopy) is an endoscopic technique that makes it possible to carry out microscopic examination of a smaller, focused spot of the mucosal layer during endoscopic procedures. Rasmussen and colleagues (2015) conducted a systematic review of the current indications and perspectives of confocal laser endomicroscopy (CLE) for IBD. Only studies reporting original clinical ...
This well established symposium will cover various aspects of modern Raman microscopy and detail the advantages of confocal Raman imaging and its applications. Scientific talks from distinguished speakers in academia and industry, the contributed talk and poster sessions, and the instrument demonstration will give the participants a deeper understanding of confocal Raman imaging.
Confocal Raman microscopy can identify particles in the 5-50 ?m range and can bridge the gap between micro-FT-IR and SEM-EDS analyses.
Scanning confocal laser microscopy (SCLM) was used to visualize fully hydrated microbial biofilms. The improved rejection of out-of-focus haze and the increased resolution of SCLM made it preferable to conventional phase microscopy for the analysis of living biofilms. The extent of image improvement was dependent on the characteristics of individual biofilms and was most apparent when films were dispersed in three dimensions, when they were thick, and when they contained a high number of cells. SCLM optical sections were amenable to quantitative computer-enhanced microscopy analyses, with minimal interference originating from overlying or underlying cell material. By using SCLM in conjunction with viable negative fluorescence staining techniques, horizontal (xy) and sagittal (xz) sections of intact biofilms of Pseudomonas aeruginosa, Pseudomonas fluorescens, and Vibrio parahaemolyticus were obtained. These optical sections were then analyzed by image-processing techniques to assess the ...
Author: Egner, A. et al.; Genre: Journal Article; Published in Print: 2002-04-01; Keywords: axial resolution; confocal microscopy; multifocal multiphoton microscopy|br/|; Title: Comparison of the axial resolution of practical Nipkow-disk confocal fluorescence microscopy with that of multifocal multiphoton microscopy: theory and experiment
TY - JOUR. T1 - Adaptations of diaphragm neuromuscular junction following inactivity. AU - Prakash, Y.s.. AU - Zhan, W. Z.. AU - Miyata, H.. AU - Sieck, Gary C. PY - 1995. Y1 - 1995. N2 - We hypothesized that differences exist in the morphological adaptations of neuromuscular junctions (NMJs) on different fiber types in response to prolonged inactivation. Two weeks of inactivity of both phrenic motoneurons and diaphragm muscle was induced by spinal cord hemitransection at C2 (spinal isolation; SI). A three-color fluorescent immunocytochemical technique, combined with laser-scanning confocal microscopy, was used to create two- (2D) and three-dimensional (3D) images of NMJs and obtain morphological information concerning: (1) innervating axons and presynaptic nerve terminals; (2) motor endplates (postsynaptic apparatus consisting of acetylcholine receptors), and (3) myosin heavy chain (MHC) phenotype of muscle fibers. In both sham controls (CTL) and SI animals, planar (2D) and surface (3D) areas ...
With the use of HRTII-RCM, we have obtained in vivo images of the central cornea in eyes affected by TSPK. Although dendritic cells were observed in the unaffected healthy corneas of the two patients with unilateral TSPK, these cells were greatly increased in number and formed aggregates in both the basal cell layer of the corneal epithelium and in Bowmans layer of the affected eyes. The dendritic cells in Bowmans layer appeared associated with the nerve fibers that form the nerve plexus of the corneal epithelium. Medical intervention reduced the numbers of these dendritic cells to normal values. With use of HRTII-RCM, we were able to observe abnormalities of cell shape and cell density in the eyes affected by TSPK. In the affected eyes, accumulation of dendritic cells was apparent in the focal lesions. Tandem scanning confocal microscopy of eyes affected by TSPK previously revealed multiple highly reflective filamentous structures in Bowmans layer of the corneal epithelium, but these ...
White identified the first gene with a demonstrated role in determining synaptic specificity. He studied the role of cell-cell interaction in determining the lineage pattern, stimulating a wide field of research. In more recent work, John and his co-workers partially confirmed his earlier model of cytokinesis; they discovered genes controlling cytokinesis and found features previously thought specific to plant cell division. Recognising the potentialities of laser-scanning confocal microscopy, John built a prototype microscope: with William Bradshaw Amos he developed this into a commercially produced instrument now widely used.[5] ...
Background and study aims: Confocal laser endomicroscopy (CLE) allows mucosal barrier defects along the intestinal epithelium to be visualized in vivo during endoscopy. Training in CLE interpretation can be achieved didactically or through self-directed learning. This study aimed to compare the effectiveness of expert-led didactic with self-directed audiovisual teaching for training inexperienced analysts on how to recognize mucosal barrier defects on endoscope-based CLE (eCLE). Materials and methods: This randomized controlled study involved trainee analysts who were taught how to recognize mucosal barrier defects on eCLE either didactically or through an audiovisual clip ...
Four types of confocal microscopes are commercially available: Confocal laser scanning microscopes use multiple mirrors (typically 2 or 3 scanning linearly along the x and the y axis) to scan the laser across the sample and descan the image across a fixed pinhole and detector.. Spinning-disk (Nipkow disk) confocal microscopes use a series of moving pinholes on a disc to scan spots of light. Since a series of pinholes scans an area in parallel each pinhole is allowed to hover over a specific area for a longer amount of time thereby reducing the excitation energy needed to illuminate a sample when compared to laser scanning microscopes. Decreased excitation energy reduces photo-toxicity and photo-bleaching of a sample often making it the preferred system for imaging live cells or organisms.. Microlens enhanced or dual spinning disk confocal microscopes work under the same principles as spinning-disk confocal microscopes except a second spinning disk containing micro-lenses is placed before the ...
The Advanced Light Microscopy Facility (ALMF) is available to all workgroups of the Institute of Biology of the Humboldt-University Berlin. Providing sufficient free capacity ALFM also provides access to its equipment for other groups of the Humboldt-University Berlin and for collaborating users not associated with the Humboldt-University of Berlin. We offer various methods of light microscopy, for instance epifluorescence microscopy, confocal laser scanning microscopy (CLSM) and spinning disk confocal laser microscopy (SDM). The instruments are equipped for the following advanced techniques: ...
Intravital microscopy of leukocyte-endothelial dynamics using the Heidelberg confocal laser microscope in scleritis and allergic conjunctivitis Journal Articles Refereed ...
Confocal Raman microscopy is a high-resolution imaging technique that is widely used for the characterization of materials and specimens in terms of thei...
Ice rheology, the integrity of polar ice core records, and ice−atmosphere interactions are among the phenomena controlled by the morphology and composition of interstitial fluids threading polycrystalline ice. Herein, we investigate how ionic impurities affect such features via time-resolved confocal fluorescence microscopy of freezing electrolyte solutions doped with a pH probe. We find that the 10 μM probe accumulates into 12 μm thick glassy channels in frozen water, but it is incorporated into randomly distributed ,1 μm diameter inclusions in freezing 1 mM NaCl. We infer that morphology is largely determined by the dynamic instabilities generated upon advancing ice by the rejected solute, rather than by thermodynamics. The protracted alkalinization of the fluid inclusions reveals that the excess negative charge generated by the preferential incorporation of Cl^− over Na^+ in ice is neutralized by the seepage of the OH^− slowly produced via H_2O → H^+ + OH^− thermal dissociation. ...
The |i|Journal of Biomedical Optics|/i| (JBO) is an open access journal that publishes peer-reviewed papers on the use of novel optical systems and techniques for improved health care and biomedical research.
Gastroenterology Research and Practice is a peer-reviewed, Open Access journal that provides a forum for researchers and clinicians working in the areas of gastroenterology, hepatology, pancreas and biliary, and related cancers. The journal welcomes submissions on the physiology, pathophysiology, etiology, diagnosis, and therapy of gastrointestinal diseases.
Author(s): Oegema, K; Whitfield, WG; Alberts, B | Abstract: CP190, a protein of 1,096 amino acids from Drosophila melanogaster, oscillates in a cell cycle-specific manner between the nucleus during interphase, and the centrosome during mitosis. To characterize the regions of CP190 responsible for its dynamic behavior, we injected rhodamine-labeled fusion proteins spanning most of CP190 into early Drosophila embryos, where their localizations were characterized using time-lapse fluorescence confocal microscopy. A single bipartite 19-amino acid nuclear localization signal was detected that causes nuclear localization. Robust centrosomal localization is conferred by a separate region of 124 amino acids; two adjacent, nonoverlapping fusion proteins containing distinct portions of this region show weaker centrosomal localization. Fusion proteins that contain both nuclear and centrosomal localization sequences oscillate between the nucleus and the centrosome in a manner identical to native CP190. Fusion
Green fluorescent protein (GFP) is a molecular marker used in synthetic biology to report gene expression and successful transformation, and in fluorescence confocal microscopy to label or characterise proteins and pathways. Equipment used to measure its expression is usually present in most laboratories, making it an accessible marker to use. Fluorescein, a bright green fluorophore, and its derivatives for example Fluorescein Isothiocyanate (FITC), have high absorptivity, excellent fluorescence and good water solubility, characteristics making them the most common fluorescent reagents for biological research (ThermoFisher Scientific). Together, these markers can be used as powerful tools for synthetic biology measurement and analysis, especially when used in technical standards. Without such standards, it can be difficult to obtain reliable and repeatable results, as cells are inherently complex biological systems that give differing results depending on the methods and machinery used. Absolute ...
Damage to peripheral nerves caused during a surgical intervention often results in function loss. Fluorescence imaging has the potential to improve intraoperative identification and preservation of these structures. However, only very few nerve targeting agents are available. This study describes the in vivo nerve staining capabilities of locally administered fluorescent lectin-analogues. To this end WGA, PNA, PHA-L and LEL were functionalized with Cy5 (λex max 640 nm; λem max 680 nm). Transfer of these imaging agents along the sciatic nerve was evaluated in Thy1-YFP mice (n = 12) after intramuscular injection. Migration from the injection site was assessed in vivo using a laboratory fluorescence scanner and ex vivo via fluorescence confocal microscopy. All four lectins showed retrograde movement and staining of the epineurium with a signal-to-muscle ratio of around two. On average, the longest transfer distance was obtained with WGA-Cy5 (0.95 cm). Since WGA also gave minimal uptake in the lymphatic
AIM: Endothelial glycocalyx (eGLX), the inner lining of glomerular capillaries, regulates glomerular albumin permselectivity and is damaged in diabetes. Recently, it has been shown that the eGLX can be modified and repaired by different substances, in particular angiopoietinl (Angl) and hydrocortisone. We hypothesise that direct restoration of eGLX in the glomerulus will improve glomerular permeability in diabetes. To be able to test this hypothesis Angl and hydrocortisone were tested in vitro, then the best candidate was chosen for further ex vivo studies. To be able to test this compound ex vivo I also designed, characterised and validated a new albumin permeability assay based on quantitative fluorescence confocal microscopy. RESULTS: Angl was found to induce the most significant eGLX changes in vitro. Angl increased the expression of surface glycosaminoglycans (GAG) and significantly increased their release into the media of human conditionally immortalised glomerular endothelial cells ...
The Human Protein Atlas portal is an open-access database supplying protein and RNA expression data for postnatal human tissues and cell lines.. The protein expression data includes millions of high-resolution images showing the spatial distribution of proteins in 46 different postnatal normal human tissues, 20 different cancerous tissues, and 47 different human cell lines. For a large fraction of proteins, a results of a protein array assay and an immunofluorescence-based confocal microscopy image are also provided. In LifeMap database, only data from normal tissues are presented. Additionally, only genes with moderate to strong staining intensity and high confidence level are shown (described as supportive/high reliability; see website for further information about the calculation methods).. This experiment card only contains the protein expression data. See RNA expression data in Large Scale Dataset: Human Protein Atlas - RNA Sequencing Data.. ...
The Human Protein Atlas portal is an open-access database supplying protein and RNA expression data for postnatal human tissues and cell lines.. The protein expression data includes millions of high-resolution images showing the spatial distribution of proteins in 46 different postnatal normal human tissues, 20 different cancerous tissues, and 47 different human cell lines. For a large fraction of proteins, a results of a protein array assay and an immunofluorescence-based confocal microscopy image are also provided. In LifeMap database, only data from normal tissues are presented. Additionally, only genes with moderate to strong staining intensity and high confidence level are shown (described as supportive/high reliability; see website for further information about the calculation methods).. This experiment card only contains the protein expression data. See RNA expression data in Large Scale Dataset: Human Protein Atlas - RNA Sequencing Data.. ...
These are overlaid confocal microscopy images of E-cadherin, Actin and DAPI staining in fibronectin-depleted MCF-10A-Ca1h (Ca1h) breast cancer cells. In the article, beginning on page 1579, Shinde and colleagues demonstrate that wild type Ca1h cells are not metastatic, express high amounts of intracellular fibronectin, and display a very mesenchymal phenotype. In contrast, when fibronectin is depleted from the Ca1h cells they undergo a mesenchymal-epithelial transition (as shown) characterized by a return of junctional E-cadherin expression and an enhanced ability to form pulmonary tumors following tail vein injection. As opposed to the tumor promoting role of fibronectin in the extracellular matrix, this study suggests that when epithelial-derived carcinoma cells undergo epithelial-mesenchymal transition, constitutive expression of fibronectin stabilizes their mesenchymal phenotype and inhibits the cell autonomous ability of those cells to complete the final steps of the metastatic process. ...
Expression and localization of proteins in a large variety of normal human tissues, cancer cells and cell lines with the aid of immunohistochemistry (IHC) images and immunofluorescence (IF) confocal microscopy images.. ...
Characterisation of cornerstone adhesions. a Spinning disk images of hPSC plated on vitronectin (VTN) and stained for F-actin, paxillin, DAPI, and the pluripotency marker Nanog. Scale bar 10 μm. b Structured illumination microscopy images of hPSC plated on VTN and stained for filamentous actin (F-actin) and paxillin. Scale bar 10 μm. The white square highlights a region of interest (ROI), which is magnified. c-f Live-cell imaging of endogenously tagged paxillin in hPSC. Images were acquired every minute using a spinning disk microscope. Colonies were recorded for at least 105 min (n = 16 independent colonies from three biologically independent experiments). A representative video is available as supplementary information (Supplementary Movie 1). c A representative image is displayed (ROI highlighted by white square and magnified; size, 30 µm) in addition to a mask image highlighting paxillin-positive adhesions (colony edge FA, green; colony centre FA, magenta) and merged images depicting ...
a) Illustration of the orientation and the dimension of the x-z optical section transverse to the capillary in the mesentery. The mesentery is arranged horizontally on the surface of the coverslip that forms the base of the observation stage. The optical section is composed of multiple line scans of the laser along the x axis, collected as the objective is lowered along the z axis. (b) When the fluorescein-dyed α-lactalbumin is perfused into the vessel lumen, the dye will travel across the vessel wall and accumulate in the tissue space surrounding the vessel. (c) An x-z confocal image after 13s perfusion of FITC-α-lactalbumin. It shows that the fluorescence dye fills the microvessel lumen and spreads in the surrounding tissue [schematic in (b)]. ...
Glutamate induces de novo growth of functional spines in developing cortex Nature 474, 7349 (2011). doi:10.1038/nature09986 Authors: Hyung-Bae Kwon & Bernardo L. Sabatini. Mature cortical pyramidal neurons receive excitatory inputs onto small protrusions emanating from their dendrites called spines. Spines undergo activity-dependent remodelling, stabilization and pruning during development, and similar structural changes can be triggered by learning and changes in sensory experiences. However, the biochemical triggers and mechanisms of de novo spine formation in the developing brain and the functional significance of new spines to neuronal connectivity are largely unknown. Here we develop an approach to induce and monitor de novo spine formation in real time using combined two-photon laser-scanning microscopy and two-photon laser uncaging of glutamate. Our data demonstrate that, in mouse cortical layer 2/3 pyramidal neurons, glutamate is sufficient to trigger de novo spine growth from the ...
METHODS: The experimental animal species were small adult pigs (n=10; body weight 30-35 kg; 4-5 months old) used for their relative physiological and metabolic resemblance to man. The following experimental methods were used for diagnostic verification of gastrointestinal lesions (damage scale: 1 - erosions, red spots, inflammatory infiltration, 2 - single ulcers, 3 - strings of ulcers): endoscopic image for the diagnostics of gastro-duodenal segment (in vivo conditions), confocal laser microscopy (ex vivo imaging) and optical light microscopy (in vitro), small intestinal imaging by means of wireless capsule enteroscopy (in vivo), macroscopic findings and optical light microscopy (after animal sacrifice ...
Confocal laser endomicroscopy (pCLE) provides real-time histologic imaging of human tissues at a depth of 60-70 μm during endoscopy. pCLE of the extrahepatic bile duct after fluorescein injection demonstrated a reticular pattern within fluorescein-filled sinuses that had no known anatomical correlate. Freezing biopsy tissue before fixation preserved the anatomy of this structure, demonstrating that it is part of the submucosa and a previously unappreciated fluid-filled interstitial space, draining to lymph nodes and supported by a complex network of thick collagen bundles. These bundles are intermittently lined on one side by fibroblast-like cells that stain with endothelial markers and vimentin, although there is a highly unusual and extensive unlined interface between the matrix proteins of the bundles and the surrounding fluid. We observed similar structures in numerous tissues that are subject to intermittent or rhythmic compression, including the submucosae of the entire gastrointestinal tract and
Hi all,I have a modified RO (reverse osmosis) membrane and I want to check biofilm growth on the membrane surface. For this purpose, Im using two bacterial strains (E.coli and Pseudomonas aeruginosa) in my system. I want to see and analyze (using confocal microscope) which strain is causing more biofilm on the membrane surface when grown in mixed culture. But, since Im not microbiologist I dont really know which stains I have to use. - Do I have to stain each of the bacteria before mixing and growing them in the system (e.g. red for E.coli and green for P.aeruginosa)? - Are the stains (e.g. SYTO 59) permanent stains? Thanks a lot for your help! Ruth ...
Cell Culture, Drug Treatment, and Cytotoxicity Assay. Human hepatoma HepG2 cells were purchased from American Type Culture Collection (Manassas, VA) and cultured according to a method described previously (Chen et al., 2000). Ketamine was dissolved in phosphate-buffered saline (0.14 M NaCl, 2.6 mM KCl, 8 mM Na2HPO4, and 1.5 mM KH2PO4). Concentrations of ketamine (≤100 μM), which correspond to clinical plasma concentrations (Domino et al., 1982; Grant et al., 1983), were chosen as the treated dosages in this study. To avoid drug interaction, when HepG2 cells were exposed to ketamine, the culture medium did not contain serum and antibiotics. Levels of γ-glutamyltranspeptidase and lactate dehydrogenase in the culture medium were measured to evaluate the cytotoxicity of ketamine to HepG2 cells as described previously (Wu et al., 2007).. Confocal Microscopic Analyses of F-Actin and Microtubular Cytoskeletons. The F-actin and microtubular cytoskeletons in HepG2 cells were visualized as described ...
Many new drugs are being developed that may slow PD progression. Currently, the main way to measure whether these drugs are working is to examine individuals at regular intervals but this examination varies greatly from one day to the next, such that it takes hundreds of participants over several years to find out if a new drug works. We hope to use a simple eye test to measure whether these drugs can slow nerve damage associated with PD. By using a more reliable and accurate measure, we hope to make an earlier diagnosis possible and to improve clinical trials to speed up the approval of new drugs.. Next Steps for Development: ...
A Quick Overview of Cancer Protocols and Upcoming Changes in Tumor Staging by Julie Ann Walby, M.D.. MUSE and Confocal Laser Endomicroscopy by Luis Brandi, M.D.. ...
Precision Raman Equipment for Confocal Raman Spectroscopy. Find out about our Raman Spectroscopy Equipment, including Confocal Raman Microscope.
분광기는 기초과학인 물리학과 화학에서는 기본장비로 활용되지만 응용분야인 바이오,나노,환경,재료 등 시험평가 부분에서는 Raman(라만),Photo Luminescence(광발광),Fluorescence(형광발광)시스템으로 핵심적인 역할을 한다. 이에 따라 동우옵트론은 마이크로 라만(Micro Raman) 공촛점 라만(Confocal Raman) PL/Raman mapping 시스템을 개발,이를 국내외에 공급함으로써 여러분야 발전에 이바지 하고 있다.. ...
Raman spectroscopy has long been applied in geoscience, for example for the identification and characterization of minerals, or in the observation of mineral phase transitions in high and ultra-high pressure/temperature experiments. In most cases because measurements have been carried out in a micro-Raman set up very little detail on the spatial distribution and association components or mineral phases, or chemical variation could be observed. In this application note from WITec, by means of Con
The |i|Journal of Biomedical Optics|/i| (JBO) publishes peer-reviewed papers on the use of novel optical systems and techniques for improved health care and biomedical research.
The Zeiss LSM 710 Confocal Laser Scanning Microscope. Laboratories for Molecular Medicine, Room 111. The Zeiss LSM 710 confocal laser scanning microscope is designed for imaging fluorescence, either in fixed or live samples. A pinhole removes the out-of-focus light and allows investigators to acquire thin optical sections at various focal planes. Stacks of images can be acquired for 3D visualization.. The confocal module is mounted on a Zeiss Axio Observer Z1 inverted microscope with high quality objectives (listed below). A heated stage is available for imaging live samples at 37°C. The microscope has multiple lasers for excitation at 405, 458, 488, 514, 561, 594, and 633 nm, and is equipped with a 34-channel QUASAR detector, which can be set to collect emitted light of any wavelength within the visible spectrum. The QUASAR detector is capable of acquiring spectral information and its flexibility allows imaging of a broad range of fluorescent indicators, including the Alexa-Fluors, ...