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

Cancer Biology

Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism Authors:  Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 6905, Q&A: 0
Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...
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Cell Biology

Transmission Electron Microscopy for Analysis of Mitochondria in Mouse Skeletal Muscle Authors:  Joseph D. McMillan and Michael A. Eisenback, date: 05/20/2018, view: 2333, Q&A: 0
Skeletal muscle is the most abundant tissue in the human body and regulates a variety of functions including locomotion and whole-body metabolism. Skeletal muscle has a plethora of mitochondria, the organelles that are essential for aerobic generation of ATP which provides the chemical energy to fuel vital functions such as contraction. The number ...
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Ex vivo Follicle Rupture and in situ Zymography in Drosophila Authors:  Elizabeth M Knapp, Lylah D Deady and Jianjun Sun, date: 05/20/2018, view: 1090, Q&A: 0
Ovulation, the process of releasing a mature oocyte from the ovary, is crucial for animal reproduction. In order for the process of ovulation to occur, a follicle must be fully matured and signaled to rupture from the ovary. During follicle rupture in both mammals and Drosophila, somatic follicle cells are enzymatically degraded to allow ...
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Intra-amniotic Injection of Mouse Embryos Authors:  Liyuan Cui, Peng Zou, Qingshuo Meng, Lu Lu and Jiayi Zhang, date: 05/20/2018, view: 960, Q&A: 0
Recent outbreaks of infectious neuro-developmental diseases such as congenital Zika syndrome - have led to a demand for prognosis data from animal models. We developed an intra-amniotic injection mice model that allows Zika virus (ZIKV) infected mice to grow to puberty. In this system, ZIKV is injected into the amniotic fluid of pregnant mice and ...
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Developmental Biology

In vitro Explant Cultures to Interrogate Signaling Pathways that Regulate Mouse Lung Development Authors:  Changfu Yao, Gianni Carraro and Barry R. Stripp, date: 05/20/2018, view: 1166, Q&A: 0
Early mouse lung development, including specification of primordia, patterning of early endoderm and determination of regional progenitor cell fates, is tightly regulated. The ability to culture explanted embryonic lung tissue provides a tractable model to study cellular interactions and paracrine factors that regulate these processes. We provide ...
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Immunology

High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level Authors:  Thomas Luh, Kimberly Lucero, Wenji Ma, Jaeyop Lee, Yu Jerry Zhou, Yufeng Shen and Kang Liu, date: 05/20/2018, view: 1303, Q&A: 0
The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to evaluate high-dimensional transcriptomic and proteomic data (relating to cellular mRNA and protein), ...
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Microbiology

In vitro Analysis of Ubiquitin-like Protein Modification in Archaea Authors:  Xian Fu, Zachary Adams and Julie A. Maupin-Furlow, date: 05/20/2018, view: 1034, Q&A: 0
The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of ...
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Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells Authors:  Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 970, Q&A: 0
Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...
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Virucidal and Neutralizing Activity Tests for Antiviral Substances and Antibodies Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/20/2018, view: 1390, Q&A: 0
In a narrow definition, virucidal activity represents the activity by which to interact with and physically disrupt viral particles. In a broad definition, it includes the activity by which to functionally inhibit (neutralize) viral infectivity without apparent morphological alterations of the viral particles. The viral infectivity can be measured ...
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Enzymatic Activity Assay for Invertase in Synechocystis Cells Authors:  Xiaoming Tan, Kuo Song and Xuefeng Lu, date: 05/20/2018, view: 1016, Q&A: 0
Invertase can catalyze the hydrolysis of sucrose, and is widely distributed in cells of cyanobacteria and plants. Being responsible for the first step for sucrose metabolism, invertase plays important physiological roles and its enzymatic activity is frequently needed to be determined. All the methods for determination of the invertase activity ...
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Molecular Biology

Ectopic Gene Expression in Macrophages Using in vitro Transcribed mRNA Authors:  Pallavi Chandra and Jennifer A. Philips, date: 05/20/2018, view: 1130, Q&A: 0
Macrophages are immune cells that contribute to host defense through various mechanisms including phagocytosis and antigen presentation. Their antimicrobial capacity is subverted by clinically important intracellular pathogens such as Mycobacterium tuberculosis. The study of host-pathogen interactions using these cells is therefore of ...
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Neuroscience

Sociability and Social Novelty Preference Tests Using a U-shaped Two-choice Field Authors:  Eun-Hwa Lee, Jin-Young Park, Yunjin Lee and Pyung-Lim  Han, date: 05/20/2018, view: 1197, Q&A: 0
We developed sociability tests based on use of a U-shaped two-choice field to read out behavioral states of sociability in rodents. The U-shaped two-choice field is a modified open field that is partially partitioned with a wall projecting to the central point, resulting in two symmetrical rectangular fields, each containing closed and open square ...
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Induction of Photothrombotic Stroke in the Sensorimotor Cortex of Rats and Preparation of Tissue for Analysis of Stroke Volume and Topographical Cortical Localization of Ischemic Infarct Authors:  Anna M. Wiersma and Ian R. Winship, date: 05/20/2018, view: 1737, Q&A: 0
The photothrombotic model of stroke is commonly used in research as it allows the ischemic infarct to be targeted to specific regions of the cortex with high reproducibility and well-defined infarct borders. Unlike other models of stroke, photothrombosis allows the precise size and location of infarct to be tightly controlled with minimal surgical ...
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In vivo Use of Dextran-based Anterograde Cortical Tracers to Assess the Integrity of the Cortical Spinal Tract Authors:  Anna M. Wiersma and Ian R. Winship, date: 05/20/2018, view: 1171, Q&A: 0
When injected into the motor cortex of rats, anterograde tracers label fibers of the associated descending corticospinal tract (CST) that originate from pyramidal neurons in the tracer-injected cortex. These fibers can be assessed at the level of the spinal cord to determine the integrity of the descending CST and the spatial distribution of axons ...
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Plant Science

Quantification of Salicylic Acid (SA) and SA-glucosides in Arabidopsis thaliana Authors:  Valérie Allasia, Benoit Industri, Michel Ponchet, Michaël Quentin, Bruno Favery and Harald Keller, date: 05/20/2018, view: 1729, Q&A: 0
Homeostasis between the cytoplasmic plant hormone salicylic acid (SA) and its’ inactive, vacuolar storage forms, SA-2-O-β-D-glucoside (SAG) and SA-β-D-Glucose Ester (SGE), regulates the fine-tuning of defense responses to biotrophic pathogens in Arabidopsis thaliana. This protocol describes a simplified, optimized procedure to ...
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Rapid Screening and Evaluation of Maize Seedling Resistance to Stalk Rot Caused by Fusarium spp. Authors:  Yali Sun, Xinsen Ruan, Liang Ma, Fang Wang and Xiquan Gao, date: 05/20/2018, view: 1229, Q&A: 0
Corn stalk rot caused by Fusarium spp., a genus of soil-borne fungal pathogens, has become a major concern of maize production. This disease normally causes significant reduction of maize yield and quality worldwide. The field assay for identifying stalk rot resistance using adult plants is largely relying on large population, yet ...
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Stem Cell

Hair Follicle Stem Cell Isolation and Expansion Authors:  Mindy Call, Ewa Anna Meyer, Winston W. Kao, Friedrich E. Kruse and Ursula Schlӧtzer-Schrehardt, date: 05/20/2018, view: 1350, Q&A: 0
Stem cells are widely used for numerous clinical applications including limbal stem cell deficiency. Stem cell derived from the bulge region of the hair follicle have the ability to differentiate into a variety of cell types including interfollicular epidermis, hair follicle structures, sebaceous glands and corneal epithelial cells when provided ...
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Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier Authors:  Mindy Call, Ewa Anna Meyer, Winston W. Kao, Friedrich E. Kruse and Ursula Schlӧtzer-Schrehardt, date: 05/20/2018, view: 1039, Q&A: 0
The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice ...
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Osteoblast Sorting and Intracellular Staining of CXCL12 Authors:  Weihuan Wang, Gurnoor Majhail, Cui Lui and Lan Zhou, date: 05/20/2018, view: 1180, Q&A: 0
Osteoblasts are bone marrow endosteum-lining niche cells playing important roles in the regulation of hematopoietic stem cells by secreting factors and cell adhesion molecules. Characterization of primary osteoblasts has been achieved through culture of outgrowth of collagenase treated bone. Immunophenotyping and flow-based analysis of long bone ...
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A Method to Injure, Dissect and Image Indirect Flight Muscle of Drosophila Authors:  Kunal Chakraborty, K. VijayRaghavan and Rajesh Gunage, date: 05/20/2018, view: 1667, Q&A: 0
Inducing an injury specifically to Drosophila flight muscles is a difficult task, owing to the small size of the muscles and the presence of the cuticle. The protocol described below provides an easy and reproducible method to induce injury in the Drosophila flight muscles.
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