• Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

Laminarin Quantification in Microalgae with Enzymes from Marine Microbes Authors:  Stefan Becker and Jan-Hendrik Hehemann, date: 04/20/2018, view: 1376, Q&A: 0
The marine beta-glucan laminarin is an abundant storage polysaccharide in microalgae. High production rates and rapid digestion by heterotrophic bacteria turn laminarin into an ideal carbon and energy source, and it is therefore a key player in the marine carbon cycle. As a main storage glucan laminarin also plays a central role in the energy ...
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Cancer Biology

FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe Authors:  Shengli Dong and Suresh K Alahari, date: 04/20/2018, view: 1876, Q&A: 0
This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells in vitro. The method is a slightly modified and updated version as previously described (Dong et al., 2017). Briefly, the target cells are incubated with the fluorescently tagged ...
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Generation of Luciferase-expressing Tumor Cell Lines Authors:  Todd V. Brennan, Liwen Lin, Xiaopei Huang and Yiping Yang, date: 04/20/2018, view: 3024, Q&A: 1
Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track tumor development and quantify tumor burden in vivo. For this purpose, the introduction of genes that confer tumors with bioluminescent ...
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Cell Biology

A Method for Extracting the Nuclear Scaffold from the Chromatin Network Authors:  Junjie Chen, Boon Heng Dennis Teo and Jinhua Lu, date: 04/20/2018, view: 1290, Q&A: 0
Each cell contains many large DNA polymers packed in a nucleus of approx. 10 μm in diameter. With histones, these DNA polymers are known to form chromatins. How chromatins further compact in the nucleus is unclear but it inevitably depends on an extensive non-chromatin nuclear scaffold. Imaging of endogenous chromatin network and the complementary ...
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Immunology

3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells Authors:  Lingli Long, Mingzhu Yin and Wang Min, date: 04/20/2018, view: 1966, Q&A: 0
Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated ...
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Microbiology

Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 1258, Q&A: 0
Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
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Quantification of Bacterial Twitching Motility in Dense Colonies Using Transmitted Light Microscopy and Computational Image Analysis Authors:  Benjamin Smith, Jianfang Li, Matteo Metruccio, Stephanie Wan, David Evans and Suzanne Fleiszig, date: 04/20/2018, view: 1054, Q&A: 0
A method was developed to allow the quantification and mapping of relative bacterial twitching motility in dense samples, where tracking of individual bacteria was not feasible. In this approach, movies of bacterial films were acquired using differential interference contrast microscopy (DIC), and bacterial motility was then indirectly quantified ...
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Extraction of Small Molecules from Fecal Samples and Testing of Their Activity on Microbial Physiology Authors:  Eduardo S. Alves, Rosana B. R. Ferreira and L. Caetano M. Antunes, date: 04/20/2018, view: 1079, Q&A: 0
The human body is colonized by vast communities of microbes, collectively known as microbiota, or microbiome. Although microbes colonize every surface of our bodies that is exposed to the external environment, the biggest collection of microbes colonizing humans and other mammals can be found in the gastrointestinal tract. Given the fact that the ...
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Determination of Intracellular Osmolytes in Cyanobacterial Cells Authors:  Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 965, Q&A: 0
Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...
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Host-regulated Hepatitis B Virus Capsid Assembly in a Mammalian Cell-free System Authors:  Kuancheng Liu and Jianming Hu, date: 04/20/2018, view: 1284, Q&A: 0
The hepatitis B virus (HBV) is an important global human pathogen and represents a major cause of hepatitis, liver cirrhosis and liver cancer. The HBV capsid is composed of multiple copies of a single viral protein, the capsid or core protein (HBc), plays multiple roles in the viral life cycle, and has emerged recently as a major target for ...
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Method for CRISPR/Cas9 Mutagenesis in Candida albicans Authors:  Neta Dean and Henry Ng, date: 04/20/2018, view: 2361, Q&A: 0
Candida albicans is the most prevalent and important human fungal pathogen. The advent of CRISPR as a means of gene editing has greatly facilitated genetic analysis in C. albicans. Here, we describe a detailed step-by-step procedure to construct and analyze C. albicans deletion mutants. This protocol uses plasmids that ...
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Metal-tagging Transmission Electron Microscopy for Localisation of Tombusvirus Replication Compartments in Yeast Authors:  Isabel Fernández de Castro and Cristina Risco, date: 04/20/2018, view: 1160, Q&A: 0
Positive-stranded (+) RNA viruses are intracellular pathogens in humans, animals and plants. To build viral replicase complexes (VRCs) viruses manipulate lipid flows and reorganize subcellular membranes. Redesigned membranes concentrate viral and host factors and create an environment that facilitates the formation of VRCs within replication ...
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Molecular Biology

Generation of microRNA Sponge Library Author:  Sebastian Herzog, date: 04/20/2018, view: 1080, Q&A: 0
This protocol describes the generation and functional validation of microRNA (miRNA) sponge or decoy constructs. When expressed from a strong promoter, these transcripts can sequester specific miRNA:RISC complexes, thereby resulting in a derepression of endogenous target mRNA. Hence, cells expressing such sponges display a partial or full miRNA ...
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Neuroscience

Electrophysiological Recordings of Evoked End-Plate Potential on Murine Neuro-muscular Synapse Preparations Authors:  Giulia Zanetti, Samuele Negro, Aram Megighian and Marco Pirazzini, date: 04/20/2018, view: 1497, Q&A: 0
Neuromuscular junction (NMJ) is the specialized chemical synapse that mediates the transmission of the electrical impulse running along motor neuron axons to skeletal muscle fibers. NMJ is the best characterized chemical synapse and its study along many years of research has provided most of the general knowledge of synapse development, structure ...
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Testing Effects of Chronic Chemogenetic Neuronal Stimulation on Energy Balance by Indirect Calorimetry Authors:  Sangho Yu and Heike Münzberg, date: 04/20/2018, view: 979, Q&A: 0
The fundamental of neuroscience is to connect the firing of neurons to physiological and behavioral outcomes. Chemogenetics enables researchers to control the activity of a genetically defined population of neurons in vivo through the expression of designer receptor exclusively activated by designer drug (DREADD) in specific neurons and ...
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Magnetic Resonance Imaging and Histopathological Visualization of Human Dural Lymphatic Vessels Authors:  Seung-Kwon Ha, Govind Nair, Martina Absinta, Nicholas J. Luciano and Daniel S. Reich, date: 04/20/2018, view: 1623, Q&A: 0
In this protocol, we describe a method to visualize and map dural lymphatic vessels in-vivo using magnetic resonance imaging (MRI) and ex-vivo using histopathological techniques. While MRI protocols for routine imaging of meningeal lymphatics include contrast-enhanced T2-FLAIR and T1-weighted black-blood imaging, a more specific ...
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Isolation and Maintenance of Striatal Neurons Authors:  Luana  Naia and A. Cristina Rego, date: 04/20/2018, view: 1023, Q&A: 0
Primary cultures of murine striatal neurons are widely used to explore cellular mechanisms in neurobiology, including brain diseases. Here we describe a detailed and standardized protocol to dissect and culture embryonic murine striatal neurons GABA-positive/DARPP-32-positive for 12 days in vitro, when they show good neuronal cell ...
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Plant Science

Quantification of Thrips Damage Using Ilastik and ImageJ Fiji Authors:  Isabella G. S. Visschers, Nicole M. van Dam and Janny L. Peters, date: 04/20/2018, view: 1785, Q&A: 0
Quantification of insect damage is an essential measurement for identifying resistance in plants. In screening for host plant resistance against thrips, the total damaged leaf area is used as a criterion to determine resistance levels. Here we present an objective novel method for analyzing thrips damage on leaf disc using the freely available ...
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Qualitative Analysis of Lipid Peroxidation in Plants under Multiple Stress Through Schiff’s Reagent: A Histochemical Approach Authors:  Jay Prakash Awasthi, Bedabrata Saha, Bhaben Chowardhara, Sanjenbam Sanjibia Devi, Pankaj Borgohain and Sanjib Kumar Panda, date: 04/20/2018, view: 1643, Q&A: 0
Lipid peroxidation is a physiological indicator of both biotic and abiotic stress responses, hence is often used as a biomarker to assess stress-induced cell damage or death. Here we demonstrate an easy, quick and cheap staining method to assess lipid peroxidation in plant tissues. In this methodology, Schiff’s reagent, is used to assay for ...
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In-vitro and in-planta Botrytis cinerea Inoculation Assays for Tomato Authors:  Jiajie Lian, Hongyu Han, Jiuhai Zhao and Chuanyou Li, date: 04/20/2018, view: 1779, Q&A: 0
Botrytis cinerea (B. cinerea) attacks many crops of economic importance, represents one of the most extensively studied necrotrophic pathogens. Inoculation of B. cinerea and phenotypic analysis of plant resistance are key procedures to investigate the mechanism of plant immunity. Here we describe a protocol for B. ...
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