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https://www.readbyqxmd.com/read/28230342/ultrasensitive-detection-of-aminopeptidase-n-activity-in-urine-and-cells-with-a-ratiometric-fluorescence-probe
#1
Xinyuan He, Yanhui Xu, Wen Shi, Huimin Ma
An ultrasensitive ratiometric fluorescent probe (CVN) has been designed and synthesized by incorporating alanine into the cresyl violet fluorophore. The probe shows ratiometric fluorescence response toward aminopeptidase N (APN) through the increase of fluorescent intensity ratio of 626/575 nm. The sensitivity of the probe is ultrahigh with a detection limit of 33 pg/mL, which can quantify the contents of APN in 500-fold diluted human urine samples. Furthermore, by using ratiometric fluorescence imaging, the probe reveals significantly higher contents of APN in HepG2 cells than those in LO2 cells, which has been further used to distinguish these two types of cells in mixed cocultures...
February 23, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28230071/probing-the-molecular-mechanism-of-human-soluble-guanylate-cyclase-activation-by-no-in-vitro-and-in-vivo
#2
Jie Pan, Hong Yuan, Xiaoxue Zhang, Huijuan Zhang, Qiming Xu, Yajun Zhou, Li Tan, Shingo Nagawa, Zhong-Xian Huang, Xiangshi Tan
Soluble guanylate cyclase (sGC) is a heme-containing metalloprotein in NO-sGC-cGMP signaling. NO binds to the heme of sGC to catalyze the synthesis of the second messenger cGMP, which plays a critical role in several physiological processes. However, the molecular mechanism for sGC to mediate the NO signaling remains unclear. Here fluorophore FlAsH-EDT2 and fluorescent proteins were employed to study the NO-induced sGC activation. FlAsH-EDT2 labeling study revealed that NO binding to the H-NOX domain of sGC increased the distance between H-NOX and PAS domain and the separation between H-NOX and coiled-coil domain...
February 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28229450/endogenous-nitric-oxide-formation-in-cardiac-myocytes-does-not-control-respiration-during-%C3%AE-adrenergic-stimulation
#3
Michael Kohlhaas, Alexander G Nickel, Stefanie Bergem, Barbara Casadei, Ulrich Laufs, Christoph Maack
Endothelial nitric oxide (NO) controls cardiac oxygen (O2 ) consumption in a paracrine way by slowing respiration at the mitochondrial electron transport chain. While NO synthases (NOS) are also expressed in cardiac myocytes, it is unclear whether they control respiration in an intracrine way. Furthermore, the existence of a mitochondrial NOS is controversial. Here, by combining fluorescence imaging with electrical field stimulation, the patch-clamp method and knock-out technology, we determined the sources and consequences of intracellular NO formation during workload transitions in isolated murine and guinea pig cardiac myocytes and mitochondria...
February 22, 2017: Journal of Physiology
https://www.readbyqxmd.com/read/28228854/a-postsynthetically-2-clickable-uridine-with-arabino-configuration-and-its-application-for-fluorescent-labeling-and-imaging-of-dna
#4
Heidi-Kristin Walter, Bettina Olshausen, Ute Schepers, Hans-Achim Wagenknecht
The arabino-configured analog of uridine with a propargyl group at the 2'-position was synthesized and incorporated into DNA by solid-phase chemistry. The fluorescence quantum yields of DNA strands that were postsynthetically modified by blue and green emitting cyanine-styryl dyes were improved due to the arabino-configured anchor. These oligonucleotides were used as energy transfer donors in hybrids with oligonucleotides modified with acceptor dyes that emit in the yellow-red range. These combinations give energy transfer pairs with blue-yellow, blue-red and green-red emission color changes...
2017: Beilstein Journal of Organic Chemistry
https://www.readbyqxmd.com/read/28228843/a-new-mechanism-for-efficient-hydrocarbon-electro-extraction-from-botryococcus-braunii
#5
Alexis Guionet, Bahareh Hosseini, Justin Teissié, Hidenori Akiyama, Hamid Hosseini
BACKGROUND: Recent understanding that specific algae have high hydrocarbon production potential has attracted considerable attention. Botryococcus braunii is a microalga with an extracellular hydrocarbon matrix, which makes it an appropriate green energy source. RESULTS: This study focuses on extracting oil from the microalgae matrix rather than the cells, eliminating the need for an excessive electric field to create electro-permeabilization. In such a way, technical limitations due to high extraction energy and cost can be overcome...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28228597/new-near-infrared-dyes-light-up-deep-tissue-imaging
#6
Conor L Evans
A newly synthesized class of near-infrared dyes offers strong absorption and efficient fluorescent emission for deep-tissue molecular imaging applications.
February 22, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/28228408/a-method-for-quantifying-limbal-stem-cell-niches-using-oct-imaging
#7
Michel Haagdorens, Joséphine Behaegel, Jos Rozema, Veerle Van Gerwen, Sofie Michiels, Sorcha Ní Dhubhghaill, Marie-José Tassignon, Nadia Zakaria
AIMS: To evaluate the efficacy of Fourier domain-optical coherence tomography (FD-OCT) in imaging and quantifying the limbal palisades of Vogt and to correlate these images with histological findings. METHODS: The superior and inferior limbal region of both eyes of 50 healthy volunteers were imaged by FD-OCT. Images were processed and analysed using Matlab software. In vitro immunofluorescent staining of a cadaveric donor limbus was analysed to correlate the presence of stem cells in the visualised structures...
February 22, 2017: British Journal of Ophthalmology
https://www.readbyqxmd.com/read/28228173/optimization-of-vascular-casting-for-three-dimensional-fluorescence-cryo-imaging-of-collateral-vessels-in-the-ischemic-rat-hindlimb
#8
Janina C V Schwarz, Monique G J T B van Lier, Erik N T P Bakker, Judith de Vos, Jos A E Spaan, Ed VanBavel, Maria Siebes
Development of collateral vessels, arteriogenesis, may protect against tissue ischemia, however, quantitative data on this process remain scarce. We have developed a technique for replicating the entire arterial network of ischemic rat hindlimbs in three dimensions (3D) based on vascular casting and automated sequential cryo-imaging. Various dilutions of Batson's No. 17 with methyl methacrylate were evaluated in healthy rats, with further protocol optimization in ischemic rats. Penetration of the resin into the vascular network greatly depended on dilution; the total length of casted vessels below 75 µm was 13-fold higher at 50% dilution compared with the 10% dilution...
February 2017: Microscopy and Microanalysis
https://www.readbyqxmd.com/read/28228172/experimental-parameters-leading-to-optimal-bilayers-for-total-internal-reflection-fluorescence-microscopy-visualization
#9
Elisabeth Mantil, Trinda Crippin, Anatoli Ianoul, Tyler J Avis
Supported lipid bilayer systems were evaluated following various experimental procedures in an effort to determine their appropriateness for visualization using total internal reflection fluorescence (TIRF) microscopy. The incorporation and distribution of Texas Red® 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (TR-DHPE) was studied when incorporated into bilayers of variable lipid composition using different forms of mechanical shearing. Results showed that 0.8 mol% TR-DHPE provides the most optimum TIRF images...
February 2017: Microscopy and Microanalysis
https://www.readbyqxmd.com/read/28228171/quantification-of-multicellular-organization-junction-integrity-and-substrate-features-in-collective-cell-migration
#10
Adam C Canver, Alisa Morss Clyne
Quantitative analysis of multicellular organization, cell-cell junction integrity, and substrate properties is essential to understand the mechanisms underlying collective cell migration. However, spatially and temporally defining these properties is difficult within collectively migrating cell groups due to challenges in accurate cell segmentation within the monolayer. In this paper, we present Matlab®-based algorithms to spatially quantify multicellular organization (migration distance, interface roughness, and cell alignment, area, and morphology), cell-cell junction integrity, and substrate features in confocal microscopy images of two-dimensional collectively migrating endothelial monolayers...
February 2017: Microscopy and Microanalysis
https://www.readbyqxmd.com/read/28228165/determining-optimal-fluorescent-agent-concentrations-in-dental-adhesive-resins-for-imaging-the-tooth-restoration-interface
#11
Odair Bim Júnior, Marco A Cebim, Maria T Atta, Camila M Machado, Luciana F Francisconi-Dos-Rios, Linda Wang
Fluorescent dyes like Rhodamine B (RB) have been used to identify the spatial distribution of adhesive restorative materials in the tooth/restoration interface. Potential effects of the addition of RB to dental adhesives were addressed in the past, but no further information is available on how to determine suitable concentrations of RB in these bonding agents for imaging in the confocal laser scanning microscope. This study provides systematical strategies for adding RB to viscous dental adhesive resins, focusing on the determination of the lowest range of dye concentrations necessary to achieve an acceptable image of the dentin/adhesive interface...
February 2017: Microscopy and Microanalysis
https://www.readbyqxmd.com/read/28227684/a-novel-wireless-wearable-fluorescence-image-guided-surgery-system
#12
Kunshan He, Yamin Mao, Jinzuo Ye, Yu An, Shixin Jiang, Chongwei Chi, Jie Tian, Kunshan He, Yamin Mao, Jinzuo Ye, Yu An, Shixin Jiang, Chongwei Chi, Jie Tian, Shixin Jiang, Jinzuo Ye, Kunshan He, Chongwei Chi, Jie Tian, Yamin Mao, Yu An
Segmentectomy using indocyanine green (ICG) has become a primary treatment option to achieve a complete resection and preserve lung function in early-stage lung cancer. However, owing to a lack of appropriate intraoperative imaging systems, it is a huge challenge for surgeons to identify the intersegmental plane during the operation, leading to poor prognosis. Thus, we developed a novel wireless wearable fluorescence image-guided surgery system (LIGHTEN) for fast and accurate identification of intersegmental planes in human patients...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227671/imquant-an-image-based-fluorescence-reader-for-quantitative-lateral-flow-immunoassays
#13
Kaushik Basak Chowdhury, Jayaraj Joseph, Jayaraman Kiruthi Vasan, Mohanasankar Sivaprakasam, Kaushik Basak Chowdhury, Jayaraj Joseph, Jayaraman Kiruthi Vasan, Mohanasankar Sivaprakasam, Jayaraman Kiruthi Vasan, Mohanasankar Sivaprakasam, Jayaraj Joseph, Kaushik Basak Chowdhury
Fluorescence immunoassays (FIA) have strong potential in enabling rapid quantitative immunological testing at point of care. Existing state of the art instruments employ laser scanning of FIA test kits, which require the use of electromechanical systems, making the instrument bulky, heavy and less reliable in the long term. There is a growing need for an extremely compact and reliable, motion-free, automated reader for accurate quantification of immunoassays. Here, we present ImQuant, an image based FIA reader with integrated monochromatic excitation source, optical filtering and real time image capture, assisted with an intuitive graphical user interface...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227372/multimodal-embryonic-imaging-using-optical-coherence-tomography-selective-plane-illumination-microscopy-and-optical-projection-tomography
#14
M Singh, C Wu, D Mayerich, M E Dickinson, I V Larina, K V Larin, M Singh, C Wu, D Mayerich, M E Dickinson, I V Larina, K V Larin, I V Larina, M Singh, K V Larin, D Mayerich, M E Dickinson, C Wu
The murine model is commonly utilized for studying developmental diseases. Different optical techniques have been developed to image mouse embryos, but each has its own set of limitations and restrictions. In this study, we compare the performance of the well-established technique of optical coherence tomography (OCT) to the relatively new methods of selective plane illumination microscopy (SPIM) and optical projection tomography (OPT) to assess murine embryonic development. OCT can provide label free high resolution images of the mouse embryo, but suffers from light attenuation that limits visualization of deeper structures...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227370/detection-and-localization-of-mouse-induced-pluripotent-stem-cell-formation-using-time-lapse-fluorescence-microscopy-images
#15
Yuan-Hsiang Chang, Hideo Yokota, Kuniya Abe, Jung Hsien Liu, Ming-Dar Tsai, Yuan-Hsiang Chang, Hideo Yokota, Kuniya Abe, Jung Hsien Liu, Ming-Dar Tsai, Yuan-Hsiang Chang, Kuniya Abe, Ming-Dar Tsai, Hideo Yokota, Jung Hsien Liu
We present an automated method for detection and localization of mouse Induced pluripotent stem (iPS) cells formation by using the fluorescence microscopy images. The differentiated cells that possibly undergo reprogramming to iPS cells can be detected by this method for screening reagents or culture conditions in iPS induction. Our method includes image preprocessing to enhance the fluorescence microscopy images, followed by cell detection to detect isolated fluorescent (reprogramming or reprogrammed iPS) cells...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227369/hybrid-segmentation-of-fluorescent-leschmania-infected-images-using-a-watersched-and-combined-region-merging-based-method
#16
F Ouertani, H Amiri, J Bettaib, R Yazidi, A Ben Salah, F Ouertani, H Amiri, J Bettaib, R Yazidi, A Ben Salah, A Ben Salah, H Amiri, J Bettaib, R Yazidi, F Ouertani
Automatic parasite segmentation in fluorescent images is of high importance as it serves as an easier and faster tool for detecting and counting parasites in each focus. In this paper we present a hybrid segmentation for the Promastigote form of Leishmania parasites in Indirect Immunofluorescence (IIF) images, combining edge and region-based techniques through the morphological algorithm of watershed. The proposed approach deals first with a pre-processing step to correct illumination non-uniformities in the fluorescence Leishmania-infected images before performing the initial segmentation by means of the watershed algorithm...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227367/statistical-identification-of-stimulus-activated-network-nodes-in-multi-neuron-voltage-sensitive-dye-optical-recordings
#17
Elham Fathiazar, Jorn Anemuller, Jutta Kretzberg, Elham Fathiazar, Jorn Anemuller, Jutta Kretzberg, Elham Fathiazar, Jutta Kretzberg, Jorn Anemuller
Voltage-Sensitive Dye (VSD) imaging is an optical imaging method that allows measuring the graded voltage changes of multiple neurons simultaneously. In neuroscience, this method is used to reveal networks of neurons involved in certain tasks. However, the recorded relative dye fluorescence changes are usually low and signals are superimposed by noise and artifacts. Therefore, establishing a reliable method to identify which cells are activated by specific stimulus conditions is the first step to identify functional networks...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28227364/a-method-to-quantify-co-localization-in-biological-images
#18
Shiwen Zhu, Roy E Welsch, Paul T Matsudaira, Shiwen Zhu, Roy E Welsch, Paul T Matsudaira, Roy E Welsch, Paul T Matsudaira, Shiwen Zhu
Quantitative co-localization analysis with fluorescent microscopy is a common approach to assess the spatial co-ordination of molecules and thus to understand their functions in biological processes. However, the co-localization analysis results might not be consistent due to various imaging conditions and different quantification methods used. We propose a novel method to separate a co-localization event into two aspects: co-occurrence and intensity correlation, which are usually combined as one parameter in other quantitative co-localization analyses...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28226849/investigation-of-the-ca-2-response-of-human-hnt-astrocytes-to-laser-removal-of-cellular-processes
#19
Brad J Raos, E Scott Graham, Charles P Unsworth, Brad J Raos, E Scott Graham, Charles P Unsworth, Charles P Unsworth, Brad J Raos, E Scott Graham
We demonstrate, for the first time, UV laser ablative microsurgery as a method for pruning astrocytic processes from live hNT astrocytic networks in vitro. Calcium fluorescence imaging was used to evaluate the cellular response to process ablation. The results showed that ablation of astrocyte processes induced an immediate increase in intracellular calcium level which propagated through the cells cytoplasm as a wave originating from the ablation site. The increased intracellular calcium dissipated from the body of the cell but remained high in the vicinity of the ablation site...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28226745/in-vivo-characterization-of-genetic-expression-of-virus-transduced-calcium-indicators-in-retinal-ganglion-cells-using-a-low-cost-funduscope
#20
Yao-Chuan Chang, Steven T Walston, Robert H Chow, James D Weiland, Yao-Chuan Chang, Steven T Walston, Robert H Chow, James D Weiland, Robert H Chow, Yao-Chuan Chang, Steven T Walston, James D Weiland
Virus-transduced calcium indicators are effective reporters of neural activity, offering the advantage of cell-specific labeling. To track the level of in vivo expression of genetically encoded calcium indicators (GECIs) in rodent retina, we developed a noninvasive imaging approach based on a custom-modified low-cost and simple fundus system that enabled us to monitor and characterize in vivo bright-field and fluorescence retinal image. The system clearly resolves individual retinal ganglion cells (RGCs) and axons...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
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