• Volume 11, 2021
  • Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011
Image Credit: BioProtoc.4080

Past Issues in 2021

Volume: 11 Issue: 13

Biochemistry

Visualization of Host Cell Kinase Activation by Viral Proteins Using GFP Fluorescence Complementation and Immunofluorescence Microscopy Authors:  Sherry T. Shu, Wing Fai Li and Thomas E. Smithgall, date: 07/05/2021, view: 1375, Q&A: 0

Non-receptor protein-tyrosine kinases regulate cellular responses to many external signals and are important drug discovery targets for cancer and infectious diseases. While many assays exist for the assessment of kinase activity in vitro, methods that report changes in tyrosine kinase activity in single cells have the potential to provide

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Yeast Lipid Extraction and Analysis by HPTLC Authors:  Dan Li, Zheng-Tan Zhang, Cheng-Wen He and Zhiping Xie, date: 07/05/2021, view: 818, Q&A: 0

The diversity of lipid structures, properties, and combinations in biological tissues makes their extraction and analysis an experimental challenge. Accordingly, even for one of the simplest single-cellular fungi, the budding yeast (Saccharomyces cerevisiae), numerous extraction and analysis protocols have been developed to separate and quantitate

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Biological Engineering

Building a Total Internal Reflection Microscope (TIRF) with Active Stabilization (Feedback SMLM) Authors:  Simao Coelho, Jongho Baek, J. Justin Gooding and Katharina Gaus, date: 07/05/2021, view: 1512, Q&A: 0

The data quality of high-resolution imaging can be markedly improved with active stabilization, which is based on feedback loops within the microscope that maintain the sample in the same location throughout the experiment. The purpose is to provide a highly accurate focus lock, therefore eliminating drift and improving localization precision.

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Biophysics

Single-molecule Fluorescence Technique to Monitor the Co-transcriptional Formation of G-quadruplex and R-loop Structures Authors:  Gunhyoung Lim and Sungchul Hohng, date: 07/05/2021, view: 1291, Q&A: 0

G-quadruplexes (GQ) and R-loops are non-canonical nucleic acid structures related to gene regulation and genome instability that can be formed during transcription; however, their formation mechanisms remain elusive. To address this question, we developed a single-molecule fluorescence technique to monitor the formation of G-quadruplex and R-loop

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OrganoPlate Micro-fluidic Microvessel Culture and Analysis Authors:  Abidemi Junaid and Thomas Hankemeier, date: 07/05/2021, view: 1182, Q&A: 0

The endothelial cells from the microvasculature are key drivers and targets of inflammatory and thrombotic processes in microvascular diseases. The study of bioactive lipids in inflammatory processes has been largely based on two-dimensional endothelial cell cultures. Three-dimensional microvessels-on-a-chip provides an opportunity to monitor the

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Cancer Biology

Monitoring Changes in the Oxidizing Milieu in the Endoplasmic Reticulum of Mammalian Cells Using HyPerER Authors:  Julia Birk, Beata Lizak, Christian Appenzeller-Herzog and Alex Odermatt, date: 07/05/2021, view: 1225, Q&A: 0

The production of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress are tightly linked. The generation of ROS can be both the cause and a consequence of ER stress pathways, and an increasing number of human diseases are characterized by tissue atrophy in response to ER stress and oxidative injury. For the assessment of modulators

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Cell Biology

A Method for Estimating the Potential Synaptic Connections Between Axons and Dendrites From 2D Neuronal Images Authors:  Carolina Tecuatl, Diek W. Wheeler and Giorgio A. Ascoli, date: 07/05/2021, view: 1279, Q&A: 0

Computational neuroscience aims to model, reproduce, and predict network dynamics for different neuronal ensembles by distilling knowledge derived from electrophysiological and morphological evidence. However, analyses and simulations often remain critically limited by the sparsity of direct experimental constraints on essential parameters, such

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Electron Tomography to Study the Three-dimensional Structure of the Reovirus Egress Pathway in Mammalian Cells Authors:  Isabel Fernández de Castro, Jose Jesús Fernández, Terence S. Dermody and Cristina Risco, date: 07/05/2021, view: 1162, Q&A: 0

Mammalian orthoreoviruses (reoviruses) are nonenveloped, double-stranded RNA viruses that replicate and assemble in cytoplasmic membranous organelles called viral inclusions (VIs). To define the cellular compartments involved in nonlytic reovirus egress, we imaged viral egress in infected, nonpolarized human brain microvascular endothelial cells

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Ethanol-induced Sedative Behavior: An Assay to Investigate Increased Dopamine Signaling in Caenorhabditis elegans Authors:  Anuradha Singh, Kavita Babu and Pratima Pandey, date: 07/05/2021, view: 609, Q&A: 0

Dopamine (DA) signaling affects locomotion, feeding, learning, and memory in C. elegans. Various assays have been developed to study the proteins involved in these behaviors; however, these assays show behavioral output only when there is a drastic change in DA levels. We designed an assay capable of observing behavioral output even with only

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Microbiology

Production, quantification, and infection of Amazonian Phlebovirus (Bunyaviridae) Authors:  Carolina Torturella Rath, Aislan de Carvalho Vivarini, Renata M. Pereira and Ulisses Gazos Lopes, date: 07/05/2021, view: 1118, Q&A: 0

Phlebotomine vectors, sand flies of the order Diptera, are known to transmit Leishmania parasites as well as RNA viruses (arboviruses) to humans. The arbovirus, Icoaraci Phlebovirus (BeAN 24262 - ICOV), used in this study was isolated from Nectomys rodents, a mammalian species that is the same natural sylvatic reservoir of Leishmania (Leishmania)

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Molecular Biology

Measurement of Transgene Copy Number in Plants Using Droplet Digital PCR Authors:  Yao-Min Cai, Quentin M. Dudley and Nicola J. Patron, date: 07/05/2021, view: 979, Q&A: 0

Transgenic plants are produced both to investigate gene function and to confer desirable traits into crops. Transgene copy number is known to influence expression levels, and consequently, phenotypes. Similarly, knowledge of transgene zygosity is desirable for making quantitative assessments of phenotype and tracking the inheritance of transgenes

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PCR-mediated One-day Synthesis of Guide RNA for the CRISPR/Cas9 System Authors:  Naim Hassan, Farhana Easmin, Keisuke Ekino and Satoshi Harashima, date: 07/05/2021, view: 1085, Q&A: 0

Nowadays, CRISPR (clustered regularly interspaced short palindromic repeats) and the CRISPR-associated protein (Cas9) system play a major role in genome editing. To target the desired sequence of the genome successfully, guide RNA (gRNA) is indispensable for the CRISPR/Cas9 system. To express gRNA, a plasmid expressing the gRNA sequence is

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Neuroscience

An Improved Method for Individual Tracking of Voluntary Wheel Running in Pair-housed Juvenile Mice Authors:  David A. Valientes, Anthony M. Raus and Autumn S. lvy, date: 07/05/2021, view: 634, Q&A: 0

Rodent cages equipped with access to a voluntary running wheel are commonly used to study the effects of aerobic physical activity on physiology and behavior. Notable discoveries in exercise neurobiology, including the key role of brain-derived neurotrophic factor (BDNF) in neural plasticity and cognition, have been made using rodents housed with

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Immunofluorescence of GFAP and TNF-α in the Mouse Hypothalamus Authors:  Prasad S. Dalvi and Denise D. Belsham, date: 07/05/2021, view: 609, Q&A: 0

Immunofluorescence is a reliable method for identifying specific proteins in neuronal and glial cell populations of the hypothalamus. Several immunofluorescence protocols are available to detect protein markers and neuropeptides in the hypothalamus; however, published methods may vary in subtle details that can potentially impact the final outcome

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Evaluating Baseline and Sensitised Heat Nociception in Adult Drosophila Authors:  Josephine N. Massingham, Olga Baron and G. Gregory Neely, date: 07/05/2021, view: 506, Q&A: 0

Chronic pain is a complex disease that affects a large proportion of the population. With little to no effective treatments currently available for patients, this malady presents a large burden to society. Drosophila melanogaster has been previously used to describe conserved molecular components of nociception in larvae and adults. However, adult

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Plant Science

Analysis of Soluble Sugar Content in Minute Quantities of Rice Tissues by GC-MS Authors:  Li-he Xu, Li-ying Xiao, Xue-qiong Xiao, Yan-nong Xiao and Gao-feng Wang, date: 07/05/2021, view: 1215, Q&A: 0

Soluble sugars play key roles in plant growth, development, and adaption to the environment. Characterizing sugar content profiling of plant tissues promotes our understanding of the mechanisms underlying these plant processes. Several technologies have been developed to quantitate soluble sugar content in plant tissues; however, it is difficult

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