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https://www.readbyqxmd.com/read/28727813/label-free-three-dimensional-imaging-of-caenorhabditis-elegans-with-visible-optical-coherence-microscopy
#1
Séverine Coquoz, Paul J Marchand, Arno Bouwens, Laurent Mouchiroud, Vincenzo Sorrentino, Daniel Szlag, Johan Auwerx, Theo Lasser
Fast, label-free, high-resolution, three-dimensional imaging platforms are crucial for high-throughput in vivo time-lapse studies of the anatomy of Caenorhabditis elegans, one of the most commonly used model organisms in biomedical research. Despite the needs, methods combining all these characteristics have been lacking. Here, we present label-free imaging of live Caenorhabditis elegans with three-dimensional sub-micrometer resolution using visible optical coherence microscopy (visOCM). visOCM is a versatile optical imaging method which we introduced recently for tomography of cell cultures and tissue samples...
2017: PloS One
https://www.readbyqxmd.com/read/28727427/high-throughput-synthesis-and-structure-of-zeolite-zsm-43-with-two-directional-8-ring-channels
#2
Tom Willhammar, Jie Su, Yifeng Yun, Xiaodong Zou, Mobae Afeworki, Simon C Weston, Hilda B Vroman, William W Lonergan, Karl G Strohmaier
The aluminosilicate zeolite ZSM-43 (where ZSM = Zeolite Socony Mobil) was first synthesized more than 3 decades ago, but its chemical structure remained unsolved because of its poor crystallinity and small crystal size. Here we present optimization of the ZSM-43 synthesis using a high-throughput approach and subsequent structure determination by the combination of electron crystallographic methods and powder X-ray diffraction. The synthesis required the use of a combination of both inorganic (Cs(+) and K(+)) and organic (choline) structure-directing agents...
July 20, 2017: Inorganic Chemistry
https://www.readbyqxmd.com/read/28726383/marangoni-convection-assisted-single-molecule-detection-with-nanojet-surface-enhanced-raman-spectroscopy
#3
Te-Wei Chang, Xinhao Wang, Amirreza Mahigir, Georgios Veronis, Gang Logan Liu, Manas Ranjan Gartia
Many single-molecule (SM) label-free techniques such as scanning probe microscopies (SPM) and magnetic force spectroscopies (MFS) provide high resolution surface topography information, but lack chemical information. Typical surface enhanced Raman spectroscopy (SERS) systems provide chemical information on the analytes, but lack spatial resolution. In addition, a challenge in SERS sensors is to bring analytes into the so-called "hot spots" (locations where the enhancement of electromagnetic field amplitude is larger than 10(3))...
July 20, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28718848/label-free-high-throughput-holographic-screening-and-enumeration-of-tumor-cells-in-blood
#4
Dhananjay Kumar Singh, Caroline C Ahrens, Wei Li, Siva A Vanapalli
We introduce inline digital holographic microscopy (in-line DHM) as a label-free technique for detecting tumor cells in blood. The optimized DHM platform fingerprints every cell flowing through a microchannel at 10 000 cells per second, based on three features - size, maximum intensity and mean intensity. To identify tumor cells in a background of blood cells, we developed robust gating criteria using machine-learning approaches. We established classifiers from the features extracted from 100 000-cell training sets consisting of red blood cells, peripheral blood mononuclear cells and tumor cell lines...
July 18, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28718822/a-bacterial-toxin-with-analgesic-properties-hyperpolarization-of-drg-neurons-by-mycolactone
#5
Ok-Ryul Song, Han-Byul Kim, Samuel Jouny, Isabelle Ricard, Alexandre Vandeputte, Nathalie Deboosere, Estelle Marion, Christophe J Queval, Pierre Lesport, Emmanuel Bourinet, Daniel Henrion, Seog Bae Oh, Guillaume Lebon, Guillaume Sandoz, Edouard Yeramian, Laurent Marsollier, Priscille Brodin
Mycolactone, a polyketide molecule produced by Mycobacterium ulcerans, is the etiological agent of Buruli ulcer. This lipid toxin is endowed with pleiotropic effects, presents cytotoxic effects at high doses, and notably plays a pivotal role in host response upon colonization by the bacillus. Most remarkably, mycolactone displays intriguing analgesic capabilities: the toxin suppresses or alleviates the pain of the skin lesions it inflicts. We demonstrated that the analgesic capability of mycolactone was not attributable to nerve damage, but instead resulted from the triggering of a cellular pathway targeting AT₂ receptors (angiotensin II type 2 receptors; AT₂R), and leading to potassium-dependent hyperpolarization...
July 18, 2017: Toxins
https://www.readbyqxmd.com/read/28715367/microfluidic-imaging-flow-cytometry-by-asymmetric-detection-time-stretch-optical-microscopy-atom
#6
Anson H L Tang, Queenie T K Lai, Bob M F Chung, Kelvin C M Lee, Aaron T Y Mok, G K Yip, Anderson H C Shum, Kenneth K Y Wong, Kevin K Tsia
Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring...
June 28, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28711027/high-throughput-and-label-free-parasitemia-quantification-and-stage-differentiation-for-malaria-infected-red-blood-cells
#7
Xiaonan Yang, Zhuofa Chen, Jun Miao, Liwang Cui, Weihua Guan
This work reports a high throughput and label-free microfluidic cell deformability sensor for quantitative parasitemia measurement and stage determination for Plasmodium falciparum-infected red blood cells (Pf-iRBCs). The sensor relies on differentiating the RBC deformability (a mechanical biomarker) that is highly correlated with the infection status. The cell deformability is measured by evaluating the transit time when each individual RBC squeezes through a microscale constriction (cross-section ~5µm×5µm)...
July 8, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28710672/automated-3-d-detection-of-dendritic-spines-from-in-vivo-two-photon-image-stacks
#8
P K Singh, P Hernandez-Herrera, D Labate, M Papadakis
Despite the significant advances in the development of automated image analysis algorithms for the detection and extraction of neuronal structures, current software tools still have numerous limitations when it comes to the detection and analysis of dendritic spines. The problem is especially challenging in in vivo imaging, where the difficulty of extracting morphometric properties of spines is compounded by lower image resolution and contrast levels native to two-photon laser microscopy. To address this challenge, we introduce a new computational framework for the automated detection and quantitative analysis of dendritic spines in vivo multi-photon imaging...
July 14, 2017: Neuroinformatics
https://www.readbyqxmd.com/read/28704378/characterizing-properties-of-non-estrogenic-substituted-bisphenol-analogs-using-high-throughput-microscopy-and-image-analysis
#9
Adam T Szafran, Fabio Stossi, Maureen G Mancini, Cheryl L Walker, Michael A Mancini
Animal studies have linked the estrogenic properties of bisphenol A (BPA) to adverse effects on the endocrine system. Because of concerns for similar effects in humans, there is a desire to replace BPA in consumer products, and a search for BPA replacements that lack endocrine-disrupting bioactivity is ongoing. We used multiple cell-based models, including an established multi-parametric, high throughput microscopy-based platform that incorporates engineered HeLa cell lines with visible ERα- or ERβ-regulated transcription loci, to discriminate the estrogen-like and androgen-like properties of previously uncharacterized substituted bisphenol derivatives and hydroquinone...
2017: PloS One
https://www.readbyqxmd.com/read/28703127/quantitative-real-time-imaging-of-glutathione
#10
Xiqian Jiang, Jianwei Chen, Aleksandar Bajić, Chengwei Zhang, Xianzhou Song, Shaina L Carroll, Zhao-Lin Cai, Meiling Tang, Mingshan Xue, Ninghui Cheng, Christian P Schaaf, Feng Li, Kevin R MacKenzie, Allan Chris M Ferreon, Fan Xia, Meng C Wang, Mirjana Maletić-Savatić, Jin Wang
Glutathione plays many important roles in biological processes; however, the dynamic changes of glutathione concentrations in living cells remain largely unknown. Here, we report a reversible reaction-based fluorescent probe-designated as RealThiol (RT)-that can quantitatively monitor the real-time glutathione dynamics in living cells. Using RT, we observe enhanced antioxidant capability of activated neurons and dynamic glutathione changes during ferroptosis. RT is thus a versatile tool that can be used for both confocal microscopy and flow cytometry based high-throughput quantification of glutathione levels in single cells...
July 13, 2017: Nature Communications
https://www.readbyqxmd.com/read/28694064/rna-fluorescence-in-situ-hybridization-for-high-content-screening
#11
Emmanuelle Querido, Lynda Dekakra-Bellili, Pascal Chartrand
Single molecule RNA imaging using fluorescent in situ hybridization (FISH) can provide quantitative information on mRNA abundance and localization in a single cell. There is now a growing interest in screening for modifiers of RNA abundance and/or localization. For instance, microsatellite expansion within RNA can lead to toxic gain-of-function via mislocalization of these transcripts into RNA aggregate and sequestration of RNA-binding proteins. Screening for inhibitors of these RNA aggregate can be performed by high-throughput RNA FISH...
July 7, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28690929/applying-pollen-dna-metabarcoding-to-the-study-of-plant-pollinator-interactions
#12
Karen L Bell, Julie Fowler, Kevin S Burgess, Emily K Dobbs, David Gruenewald, Brice Lawley, Connor Morozumi, Berry J Brosi
PREMISE OF THE STUDY: To study pollination networks in a changing environment, we need accurate, high-throughput methods. Previous studies have shown that more highly resolved networks can be constructed by studying pollen loads taken from bees, relative to field observations. DNA metabarcoding potentially allows for faster and finer-scale taxonomic resolution of pollen compared to traditional approaches (e.g., light microscopy), but has not been applied to pollination networks. METHODS: We sampled pollen from 38 bee species collected in Florida from sites differing in forest management...
June 2017: Applications in Plant Sciences
https://www.readbyqxmd.com/read/28686620/structural-transitions-during-large-ribosomal-subunit-maturation-analyzed-by-tethered-nuclease-structure-probing-in-s-cerevisiae
#13
Gisela Pöll, Christian Müller, Malena Bodden, Fabian Teubl, Norbert Eichner, Gerhard Lehmann, Joachim Griesenbeck, Herbert Tschochner, Philipp Milkereit
Yeast large ribosomal subunit (LSU) precursors are subject to substantial changes in protein composition during their maturation due to coordinated transient interactions with a large number of ribosome biogenesis factors and due to the assembly of ribosomal proteins. These compositional changes go along with stepwise processing of LSU rRNA precursors and with specific rRNA folding events, as revealed by recent cryo-electron microscopy analyses of late nuclear and cytoplasmic LSU precursors. Here we aimed to analyze changes in the spatial rRNA surrounding of selected ribosomal proteins during yeast LSU maturation...
2017: PloS One
https://www.readbyqxmd.com/read/28678425/semi-automated-scoring-of-triple-probe-fish-in-human-sperm-using-confocal-microscopy
#14
Francesca Branch, GiaLinh Nguyen, Nicholas Porter, Heather A Young, Sheena E Martenies, Nathan McCray, Glen Deloid, Anastas Popratiloff, Melissa J Perry
Structural and numerical sperm chromosomal aberrations result from abnormal meiosis and are directly linked to infertility. Any live births that arise from aneuploid conceptuses can result in syndromes such as Kleinfelter, Turners, XYY and Edwards. Multi-probe fluorescence in situ hybridization (FISH) is commonly used to study sperm aneuploidy, however manual FISH scoring in sperm samples is labor-intensive and introduces errors. Automated scoring methods are continuously evolving. One challenging aspect for optimizing automated sperm FISH scoring has been the overlap in excitation and emission of the fluorescent probes used to enumerate the chromosomes of interest...
July 5, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28678160/lipid-bilayer-membrane-in-a-silicon-based-micron-sized-cavity-accessed-by-atomic-force-microscopy-and-electrochemical-impedance-spectroscopy
#15
Muhammad Shuja Khan, Noura Sayed Dosoky, Darayas Patel, Jeffrey Weimer, John Dalton Williams
Supported lipid bilayers (SLBs) are widely used in biophysical research to probe the functionality of biological membranes and to provide diagnoses in high throughput drug screening. Formation of SLBs at below phase transition temperature (Tm) has applications in nano-medicine research where low temperature profiles are required. Herein, we report the successful production of SLBs at above-as well as below-the Tm of the lipids in an anisotropically etched, silicon-based micro-cavity. The Si-based cavity walls exhibit controlled temperature which assist in the quick and stable formation of lipid bilayer membranes...
July 5, 2017: Biosensors
https://www.readbyqxmd.com/read/28677396/rapid-characterization-of-a-mechanically-labile-%C3%AE-helical-protein-enabled-by-efficient-site-specific-bioconjugation
#16
Robert Walder, Marc-Andre LeBlanc, William John Van Patten, Devin Edwards, Jacob A Greenberg, Ayush Adhikari, Stephen R Okoniewski, Ruby May Arana Sullan, David Rabuka, Marcelo Carlos Sousa, Thomas T Perkins
Atomic-force-microscopy (AFM)-based single-molecule force spectroscopy (SMFS) is a powerful yet accessible means to characterize mechanical properties of biomolecules. Historically, accessibility relies upon the nonspecific adhesion of biomolecules to a surface and a cantilever and, for proteins, the integration of the target protein into a polyprotein. However, this assay results in a low yield of high-quality data, defined as the complete unfolding of the polyprotein. Additionally, nonspecific surface adhesion hinders studies of α-helical proteins, which unfold at low forces and low extensions...
July 5, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28668710/accurate-virus-quantitation-using-a-scanning-transmission-electron-microscopy-stem-detector-in-a-scanning-electron-microscope
#17
Candace D Blancett, David P Fetterer, Keith A Koistinen, Elaine M Morazzani, Mitchell K Monninger, Ashley E Piper, Kathleen A Kuehl, Brian J Kearney, Sarah L Norris, Cynthia A Rossi, Pamela J Glass, Mei G Sun
A method for accurate quantitation of virus particles has long been sought, but a perfect method still eludes the scientific community. Electron Microscopy (EM) quantitation is a valuable technique because it provides direct morphology information and counts of all viral particles, whether or not they are infectious. In the past, EM negative stain quantitation methods have been cited as inaccurate, non-reproducible, and with detection limits that were too high to be useful. To improve accuracy and reproducibility, we have developed a method termed Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ), which simplifies sample preparation and uses a high throughput STEM detector in a Scanning Electron Microscope (SEM) coupled with commercially available software...
June 28, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28668355/cell-biology-of-microbes-and-pharmacology-of-antimicrobial-drugs-explored-by-atomic-force-microscopy
#18
REVIEW
Cécile Formosa-Dague, Raphaël Emmanuel Duval, Etienne Dague
Antimicrobial molecules have been used for more than 50 years now and are the basis of modern medicine. No surgery can nowdays be imagined to be performed without antibiotics; dreadful diseases like tuberculosis, leprosis, siphilys, and more broadly all microbial induced diseases, can be cured only through the use of antimicrobial treatments. However, the situation is becoming more and more complex because of the ability of microbes to adapt, develop, acquire, and share mechanisms of resistance to antimicrobial agents...
June 28, 2017: Seminars in Cell & Developmental Biology
https://www.readbyqxmd.com/read/28649406/architecture-component-and-microbiome-of-biofilm-involved-in-the-fouling-of-membrane-bioreactors
#19
Tomohiro Inaba, Tomoyuki Hori, Hidenobu Aizawa, Atsushi Ogata, Hiroshi Habe
Biofilm formation on the filtration membrane and the subsequent clogging of membrane pores (called biofouling) is one of the most persistent problems in membrane bioreactors for wastewater treatment and reclamation. Here, we investigated the structure and microbiome of fouling-related biofilms in the membrane bioreactor using non-destructive confocal reflection microscopy and high-throughput Illumina sequencing of 16S rRNA genes. Direct confocal reflection microscopy indicated that the thin biofilms were formed and maintained regardless of the increasing transmembrane pressure, which is a common indicator of membrane fouling, at low organic-loading rates...
2017: NPJ Biofilms and Microbiomes
https://www.readbyqxmd.com/read/28644811/applying-pattern-recognition-to-high-resolution-images-to-determine-cellular-signaling-status
#20
Michael F Lohrer, Darrin M Hanna, Yang Liu, Kang-Hsin Wang, Fu-Tong Liu, Ted A Laurence, Gang-Yu Liu
Two frequently used tools to acquire high-resolution images of cells are scanning electron microscopy (SEM) and atomic force microscopy (AFM). The former provides a nanometer resolution view of cellular features rapidly and with high throughput, while the latter enables visualizing hydrated and living cells. In current practice, these images are viewed by eye to determine cellular status, e.g. activated versus resting. Automatic and quantitative data analysis is lacking. This work develops an algorithm of pattern recognition that works very effectively for AFM and SEM images...
June 21, 2017: IEEE Transactions on Nanobioscience
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