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https://www.readbyqxmd.com/read/28924667/correlative-in-resin-super-resolution-fluorescence-and-electron-microscopy-of-cultured-cells
#1
Errin Johnson, Rainer Kaufmann
Correlative super-resolution light and electron microscopy (super-resolution CLEM) is a powerful and emerging tool in biological research. The practical realization of these two very different microscopy techniques with their individual requirements remains a challenging task. There is a broad range of approaches to choose from, each with their own advantages and limitations. Here, we present a detailed protocol for in-resin super-resolution CLEM of high-pressure frozen and freeze substituted cultured cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28882053/naphthoquinone-rich-onosma-visianii-clem-boraginaceae-root-extracts-induce-apoptosis-and-cell-cycle-arrest-in-hct-116-and-mda-mb-231-cancer-cell-lines
#2
Milena D Vukic, Nenad L Vukovic, Ana D Obradovic, Suzana Lj Popovic, Milan M Zaric, Predrag M Djurdjevic, Snezana D Markovic, Dejan D Baskic
In the present study, five root extracts of Onosma visianii Clem were investigated for their in vitro cytotoxic activity. On the basis of HPLC-PDA analysis, these extracts have proved to be a rich source of naphthoquinones as natural colourants for food and cosmetic industry. All investigated root extracts contain acetylshikonin, isobutyrylshikonin and α-methylbutyrylshikonin as major compounds. As the most abundant source of active compounds for antitumour therapy, acetone, chloroform and ethyl acetate extracts showed strong cytotoxic activity towards HCT-116 and MDA-MB-231 cancer cell lines...
September 7, 2017: Natural Product Research
https://www.readbyqxmd.com/read/28874723/inducing-fluorescence-of-uranyl-acetate-as-a-dual-purpose-contrast-agent-for-correlative-light-electron-microscopy-with-nanometre-precision
#3
Maarten W Tuijtel, Aat A Mulder, Clara C Posthuma, Barbara van der Hoeven, Abraham J Koster, Montserrat Bárcena, Frank G A Faas, Thomas H Sharp
Correlative light-electron microscopy (CLEM) combines the high spatial resolution of transmission electron microscopy (TEM) with the capability of fluorescence light microscopy (FLM) to locate rare or transient cellular events within a large field of view. CLEM is therefore a powerful technique to study cellular processes. Aligning images derived from both imaging modalities is a prerequisite to correlate the two microscopy data sets, and poor alignment can limit interpretability of the data. Here, we describe how uranyl acetate, a commonly-used contrast agent for TEM, can be induced to fluoresce brightly at cryogenic temperatures (-195 °C) and imaged by cryoFLM using standard filter sets...
September 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28820036/efficacy-and-safety-of-the-early-addition-of-vasopressin-to-norepinephrine-in-septic-shock
#4
Drayton A Hammond, Julia Cullen, Jacob T Painter, Kelsey McCain, Oktawia A Clem, Amy L Brotherton, Divyan Chopra, Nikhil Meena
BACKGROUND: Delays in achievement of target mean arterial pressure (MAP) have been associated with increased mortality in patients with septic shock. Vasopressin may be added to norepinephrine to raise MAP or decrease norepinephrine dosage. The purpose of this study was to determine whether early initiation of vasopressin to norepinephrine resulted in a reduced time to target MAP compared to norepinephrine monotherapy. METHODS: This retrospective cohort study compared early addition of vasopressin within 4 hours of septic shock onset to norepinephrine versus initial norepinephrine monotherapy in medically, critically ill patients with septic shock admitted from May 2014 to October 2015...
January 1, 2017: Journal of Intensive Care Medicine
https://www.readbyqxmd.com/read/28813736/evolving-pharmacologic-therapies-for-heart-failure
#5
James R Clem, Jodi Heins
No abstract text is available yet for this article.
April 2017: South Dakota Medicine: the Journal of the South Dakota State Medical Association
https://www.readbyqxmd.com/read/28781997/using-integrated-correlative-cryo-light-and-electron-microscopy-to-directly-observe-syntaphilin-immobilized-neuronal-mitochondria-in-situ
#6
Shengliu Wang, Shuoguo Li, Gang Ji, Xiaojun Huang, Fei Sun
Correlative cryo-fluorescence and cryo-electron microscopy (cryo-CLEM) system has been fast becoming a powerful technique with the advantage to allow the fluorescent labeling and direct visualization of the close-to-physiologic ultrastructure in cells at the same time, offering unique insights into the ultrastructure with specific cellular function. There have been various engineered ways to achieve cryo-CLEM including the commercial FEI iCorr system that integrates fluorescence microscope into the column of transmission electron microscope...
2017: Biophysics Reports
https://www.readbyqxmd.com/read/28770501/unsaturated-cuticular-hydrocarbons-enhance-responses-to-sex-pheromone-in-spruce-budworm-choristoneura-fumiferana
#7
P J Silk, E Eveleigh, L Roscoe, K Burgess, S Weatherby, G Leclair, P Mayo, M Brophy
The primary sex pheromone components of the female spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), are (E)- and (Z)-11-tetradecenal, produced in 95:5 ratio. However, male flight responses to calling females in a wind tunnel were faster and maintained longer than responses to any synthetic aldehyde blend. Analyses of cuticular extracts from spruce budworm adults revealed series of n-alkanes and n-monoalkenes with predominantly odd numbers of carbon atoms from C23- C29 in both sexes...
August 2, 2017: Journal of Chemical Ecology
https://www.readbyqxmd.com/read/28703019/effectiveness-and-safety-of-magnesium-replacement-in-critically-ill-patients-admitted-to-the-medical-intensive-care-unit-in-an-academic-medical-center-a-retrospective-cohort-study
#8
Drayton A Hammond, Jelena Stojakovic, Niranjan Kathe, Julie Tran, Oktawia A Clem, Kristina Erbach, Jarrod King
BACKGROUND: "Rules of thumb" for the replacement of electrolytes, including magnesium, in critical care settings are used, despite minimal empirical validation of their ability to achieve a target serum concentration. This study's purpose was to evaluate the effectiveness and safety surrounding magnesium replacement in medically, critically ill patients with mild-to-moderate hypomagnesemia. METHODS: This was a single-center, retrospective, observational evaluation of episodes of intravenous magnesium replacement ordered for patients with mild-to-moderate hypomagnesemia (1...
January 1, 2017: Journal of Intensive Care Medicine
https://www.readbyqxmd.com/read/28639149/safety-feasibility-results-and-economic-impact-of-common-interventional-procedures-in-a-low-volume-region-of-the-united-states
#9
Aaron Clem, Sami Awadallah, Zahid Amin
The aim of the study was to evaluate the safety, feasibility, and economic benefit of a high-volume operator performing common interventional procedures in a rural region of the United States (U.S.). Rural areas of the U.S., even with well-equipped cardiac catheterization laboratories, may not have a full-time pediatric interventional cardiologist (PIC); this requires patients to travel out of state even for simple interventions. Since 2005, we have adopted a policy of performing cardiac catheterizations and common interventional procedures with a visiting PIC...
June 21, 2017: Pediatric Cardiology
https://www.readbyqxmd.com/read/28618919/discontinuation-of-vasopressin-before-norepinephrine-in-the-recovery-phase-of-septic-shock
#10
Drayton A Hammond, Kelsey McCain, Jacob T Painter, Oktawia A Clem, Julia Cullen, Amy L Brotherton, Divyan Chopra, Nikhil Meena
BACKGROUND: Guidance for the discontinuation of vasopressors in the recovery phase of septic shock is limited. Norepinephrine is more easily titrated; however, septic shock is a vasopressin deficient state, which exogenous vasopressin endeavors to resolve. Discontinuation of vasopressin before norepinephrine may result in clinically significant hypotension. METHODS: This retrospective, cohort study compared discontinuation of norepinephrine and vasopressin in medically, critically ill patients in the recovery phase of septic shock from May 2014 to June 2016...
January 1, 2017: Journal of Intensive Care Medicine
https://www.readbyqxmd.com/read/28528643/a-new-method-for-cryo-sectioning-cell-monolayers-using-a-correlative-workflow
#11
Androniki Kolovou, Martin Schorb, Abul Tarafder, Carsten Sachse, Yannick Schwab, Rachel Santarella-Mellwig
Cryo-electron microscopy (cryo-EM) techniques have made a huge advancement recently, providing close to atomic resolution of the structure of protein complexes. Interestingly, this imaging technique can be performed in cells, giving access to the molecular machines in their natural context, therefore bridging structural and cell biology. However, in situ structural electron microscopy faces one major challenge, which is the ability to focus on specific subcellular regions to capture the objects of interest...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528642/minimal-resin-embedding-of-multicellular-specimens-for-targeted-fib-sem-imaging
#12
Nicole L Schieber, Pedro Machado, Sebastian M Markert, Christian Stigloher, Yannick Schwab, Anna M Steyer
Correlative light and electron microscopy (CLEM) is a powerful tool to perform ultrastructural analysis of targeted tissues or cells. The large field of view of the light microscope (LM) enables quick and efficient surveys of the whole specimen. It is also compatible with live imaging, giving access to functional assays. CLEM protocols take advantage of the features to efficiently retrace the position of targeted sites when switching from one modality to the other. They more often rely on anatomical cues that are visible both by light and electron microscopy...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528641/preserving-the-photoswitching-ability-of-standard-fluorescent-proteins-for-correlative-in-resin-super-resolution-and-electron-microscopy
#13
Errin Johnson, Rainer Kaufmann
There are many different correlative light and electron microscopy (CLEM) techniques available. The use of super-resolution microscopy in CLEM is an emerging application and while offering the obvious advantages of improved resolution in the fluorescence image, and therefore more precise correlation to electron microscopy (EM) ultrastructure, it also presents new challenges. Choice of fluorophore, method of fixation, and timing of the fluorescence imaging are critical to the success of super-resolution CLEM and the relative importance, and technical difficulty, of each of these factors depends on the type of super-resolution microscopy being employed...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528640/ec-clem-a-multidimension-multimodel-software-to-correlate-intermodal-images-with-a-focus-on-light-and-electron-microscopy
#14
Xavier Heiligenstein, Perrine Paul-Gilloteaux, Graça Raposo, Jean Salamero
Correlative light and electron microscopy (CLEM) is a scientific method covered by a broad range of techniques. The path taken to explore a scientific question is often driven both by the question and the technology available. Yet, one common step to all CLEM workflows is the registration of the multimodal images to assign a fluorescent signal to an ultrastructure. The manual relocation and registration of light microscopy and electron microscopy images can be challenging and time-consuming (Muller-Reichert & Verkade, 2014)...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528639/matrix-maps-an-intuitive-software-to-acquire-analyze-and-annotate-light-microscopy-data-for-clem
#15
Martin Schorb, Frank Sieckmann
Matrix MAPS provides an intuitive interface for acquiring light microscopy data during a correlative light and electron microscopy experiment either at room or cryogenic temperatures. First, the user graphically defines the geometry of the acquisition region on top of preview images. Multiple independent regions can then be imaged in an automated way, each with individual settings. The same interface allows the user to mark and select points or regions of interest whose coordinates can subsequently be transferred directly to the electron microscope...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528638/triclem-combining-high-precision-room-temperature-clem-with%C3%A2-cryo-fluorescence-microscopy-to-identify-very-rare-events
#16
Nicholas R Ader, Wanda Kukulski
Fiducial-based correlation of fluorescence and electron microscopy data from high-pressure frozen and resin-embedded samples allows for high-precision localization of fluorescent signals to subcellular ultrastructure. Here we introduce the triCLEM procedure to facilitate the identification of very rare events for high-precision correlation. We present a detailed protocol to screen high-pressure frozen cell monolayers on sapphire disks for very rare signals by cryo-fluorescence microscopy, relocate the cells of interest after freeze substitution and Lowicryl embedding, and perform fiducial-based correlation of the identified fluorescent signals to high-magnification electron tomograms...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528633/correlative-light-electron-microscopy-in-liquid-using%C3%A2-an-inverted-sem-asem
#17
Chikara Sato, Takaaki Kinoshita, Nassirhadjy Memtily, Mari Sato, Shoko Nishihara, Toshiko Yamazawa, Shinya Sugimoto
In atmospheric scanning electron microscope (ASEM), the inverted scanning electron microscope (SEM) observes the wet sample from below, while an optical microscope observes it from above simultaneously. The ASEM sample holder has a disposable dish shape with a silicon nitride film window at the bottom. It can be coated variously for the primary-culture of substrate-sensitive cells; primary cells were cultured in a few milliliters of culture medium in a stable incubator environment. For the inverted SEM observation, cells and the excised tissue blocks were aldehyde-fixed, immersed in radical scavenger solution, and observed at minimum electron dose...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28528631/a-fully-integrated-three-dimensional-fluorescence-to-electron-microscopy-correlative-workflow
#18
Claudia S López, Cedric Bouchet-Marquis, Christopher P Arthur, Jessica L Riesterer, Gregor Heiss, Guillaume Thibault, Lee Pullan, Sunjong Kwon, Joe W Gray
While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam-scanning electron microscopy data...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28508074/pressure-compression-of-cdse-nanoparticles-into-luminescent-nanowires
#19
Binsong Li, Kaifu Bian, Xiaowang Zhou, Ping Lu, Sheng Liu, Igal Brener, Michael Sinclair, Ting Luk, Hattie Schunk, Leanne Alarid, Paul G Clem, Zhongwu Wang, Hongyou Fan
Oriented attachment (OA) of synthetic nanocrystals is emerging as an effective means of fabricating low-dimensional nanoscale materials. However, OA relies on energetically favorable nanocrystal facets to grow nanostructured materials. Consequently, nanostructures synthesized through OA are generally limited to a specific crystal facet in their final morphology. We report our discovery that high-pressure compression can induce consolidation of spherical CdSe nanocrystal arrays, leading to unexpected one-dimensional semiconductor nanowires that do not exhibit the typical crystal facet...
May 2017: Science Advances
https://www.readbyqxmd.com/read/28481640/multiple-breath-washout-outcomes-are-sensitive-to-inflammation-and-infection-in-children-with-cystic-fibrosis
#20
Kathryn A Ramsey, Rachel E Foong, Jasmine Grdosic, Alana Harper, Billy Skoric, Charles Clem, Miriam Davis, Lidija Turkovic, Stephen M Stick, Stephanie D Davis, Sarath C Ranganathan, Graham L Hall
RATIONALE: The lung clearance index is a measure of ventilation distribution from the multiple breath washout technique. The lung clearance index is increased in the presence of lower respiratory tract inflammation and infection in infants with cystic fibrosis, however the associations during the preschool years are unknown. OBJECTIVES: We assessed the ability of the lung clearance index to detect the presence and extent of lower respiratory tract inflammation and infection in preschool children with cystic fibrosis...
May 8, 2017: Annals of the American Thoracic Society
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