• 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.4170

Current Issue in 2021

Volume: 11 Issue: 19

Biochemistry

Proximity Dependent Biotin Labelling in Zebrafish for Proteome and Interactome Profiling Authors:  Zherui Xiong, Harriet P. Lo, Kerrie-Ann McMahon, Robert G. Parton and Thomas E. Hall, date: 10/05/2021, view: 1121, Q&A: 0

Identification of protein interaction networks is key for understanding intricate biological processes, but mapping such networks is challenging with conventional biochemical methods, especially for weak or transient interactions. Proximity-dependent biotin labelling (BioID) using promiscuous biotin ligases and mass spectrometry (MS)-based

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Transient yet Robust Expression of Proteins in the Mouse Liver via Intravenous Injection of Lipid Nanoparticle-encapsulated Nucleoside-modified mRNA Authors:  Elissa Everton, Fatima Rizvi, Anna R. Smith, Mitchell Beattie, Ying Tam, Norbert Pardi, Drew Weissman and Valerie Gouon-Evans, date: 10/05/2021, view: 801, Q&A: 0

With the recent availability of the SARS-CoV-2 mRNA-based vaccines, public attention has been drawn to this new technology and how it may be applied to other indications. Temporal activation of key hepatic regenerative pathways can induce liver regeneration, overcoming the lack of donor organs for liver transplantation and ineffectiveness of

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Biological Engineering

Zebrafish Embryos as a Predictive Animal Model to Study Nanoparticle Behavior in vivo Authors:  Gabriela Arias-Alpizar, Jeroen Bussmann and Frederick Campbell, date: 10/05/2021, view: 693, Q&A: 0

A failure to fully understand the complex in vivo behavior of systemically administered nanomedicines has stymied clinical translation. To bridge this knowledge gap, new in vivo tools are needed to rapidly and accurately assess the nearly infinite array of possible nanoparticle designs. Zebrafish embryos are small, transparent, and easily

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Cancer Biology

Analysis of Leukemia Cell Metabolism through Stable Isotope Tracing in Mice Authors:  Nick van Gastel, Jessica B. Spinelli, Marcia C. Haigis and David T. Scadden, date: 10/05/2021, view: 795, Q&A: 0

Once thought to be a mere consequence of the state of a cell, intermediary metabolism is now recognized as a key regulator of mammalian cell fate and function. In addition, cell metabolism is often disturbed in malignancies such as cancer, and targeting metabolic pathways can provide new therapeutic options. Cell metabolism is mostly studied in

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Cell Biology

Isolation of Microvesicles from Human Circulating Neutrophils Authors:  Dong Zhan, Ellie McConachie, Steven W. Edwards, Helen L. Wright, Robert J. Moots and Sittisak Honsawek, date: 10/05/2021, view: 819, Q&A: 0

Neutrophil-derived microvesicles (NDMVs) are liberated by neutrophils upon cell activation by molecules. Once activated, neutrophils are primarily involved in acute inflammation; however, the microvesicles they produce are largely anti-inflammatory. NDMVs have been shown to protect cartilage during inflammatory arthritis. They exert these effects

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Fixation and Immunostaining of Endogenous Proteins or Post-translational Modifications in Caenorhabditis elegans Authors:  Robert O'Hagan and Irini Topalidou, date: 10/05/2021, view: 734, Q&A: 0

Although the advent of genetically-encoded fluorescent markers, such as the green fluorescent protein (GFP; Chalfie et al., 1994), has enabled convenient visualization of gene expression in vivo, this method is generally not effective for detecting post-translational modifications because they are not translated from DNA sequences.

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Structural and Functional Mapping of Mesenchymal Bodies Authors:  Sébastien Sart, Raphaël F.-X. Tomasi, Antoine Barizien, Gabriel Amselem, Ana Cumano and Charles N. Baroud, date: 10/05/2021, view: 1160, Q&A: 0

The formation of spheroids with mesenchymal stem/stromal cells (MSCs), mesenchymal bodies (MBs), is usually performed using bioreactors or conventional well plates. While these methods promote the formation of a large number of spheroids, they provide limited control over their structure or over the regulation of their environment. It has

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Developmental Biology

Phalloidin Staining of Actin Filaments for Visualization of Muscle Fibers in Caenorhabditis elegans Authors:  Mario Romani and Johan Auwerx, date: 10/05/2021, view: 714, Q&A: 0

Advances in C. elegans research have allowed scientists to recapitulate different human disorders, from neurodegenerative diseases to muscle dysfunction, in these nematodes. Concomitantly, the interest in visualizing organs affected by these conditions has grown, leading to the establishment of different antibody- and dye-based staining protocols

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Isolation of Primary Cytotrophoblasts From Human Placenta at Term Authors:  Arthur Colson, Christophe Louis Depoix, Corinne Hubinont and Frédéric Debiève, date: 10/05/2021, view: 349, Q&A: 0

The placenta is a multifaceted organ, fulfilling critical functions for the fetus and the mother. Therefore, it is a critical regulator of the pregnancy, and its dysfunction leads to diseases, including fetal growth restriction and preeclampsia. Studying the placenta is a difficult task since its existence is transient, and its structure is

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Immunology

Calcium Imaging in T Lymphocytes: a Protocol for Use with Genetically Encoded or Chemical Ca2+ Indicators Authors:  Amit Jairaman and Michael D. Cahalan, date: 10/05/2021, view: 517, Q&A: 0

Elevations in cytosolic calcium (Ca2+) drive a wide array of immune cell functions, including cytokine production, gene expression, and cell motility. Live-cell imaging of cells loaded with ratiometric chemical Ca2+ indicators remains the gold standard for visualization and quantification of intracellular Ca2+ signals; ratiometric imaging can be

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Measuring Total Classical Pathway and Activities of Individual Components of the Mouse Complement Pathway Author:  Wioleta M. Zelek, date: 10/05/2021, view: 326, Q&A: 0

The complement system is a central component of innate immunity, responsible for recognition and killing of bacteria by tagging invaders through opsonisation, thereby promoting phagocytosis, and by direct lysis. Complement activity is routinely measured using functional assays that utilise erythrocytes as targets. The classical pathway haemolytic

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Microbiology

Simple Methods for the Preparation of Colloidal Chitin, Cell Free Supernatant and Estimation of Laminarinase Author:  Ananthamurthy Koteshwara, date: 10/05/2021, view: 421, Q&A: 0

Colloidal chitin (CC) is a common substrate used in research work involving chitin-active enzymes (chitinases). Cell free supernatant (CFS) is prepared from fermented broth. Preparation of CC and CFS usually involve large amounts of liquid, which must be separated from the solids. This necessitates the use of a large volume centrifugation

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Molecular Biology

A High-throughput Pipeline to Determine DNA and Nucleosome Conformations by AFM Imaging Authors:  Sebastian F. Konrad, Willem Vanderlinden and Jan Lipfert, date: 10/05/2021, view: 404, Q&A: 0

Atomic force microscopy (AFM) is a powerful tool to image macromolecular complexes with nanometer resolution and exquisite single-molecule sensitivity. While AFM imaging is well-established to investigate DNA and nucleoprotein complexes, AFM studies are often limited by small datasets and manual image analysis that is slow and prone to user bias.

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Neuroscience

Protocols to Induce, Prevent, and Treat Post-traumatic Stress Disorder-like Memory in Mice: Optogenetics and Behavioral Approaches Authors:  Aline S. Al Abed, Azza Sellami, Eva-Gunnel Ducourneau, Chloé Bouarab, Aline Marighetto and Aline Desmedt, date: 10/05/2021, view: 389, Q&A: 0

One of the cardinal features of post-traumatic stress disorder (PTSD) is a paradoxical memory alteration including both emotional hypermnesia for salient trauma-related cues and amnesia for the surrounding traumatic context. Interestingly, some clinical studies have suggested that contextual amnesia would causally contribute to the PTSD-related

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Isolation, Culture, and Identification of Primary Müller Cells from Human Retina Authors:  Yingying Chen, Ting Zhang, Shaoxue Zeng, Michelle Yam, Mark C. Gillies and Ling Zhu, date: 10/05/2021, view: 408, Q&A: 0

Müller cells, the major glial cells of the retina, play vital roles in maintaining redox homeostasis and retinal metabolism. An immortalized human Müller cell line (MIO-M1) is widely used as an in vitro model to study Müller cells’ function, but they may not be exactly the same as primarily cultured human Müller cells. The use of human

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Plant Science

Dark Respiration Measurement from Arabidopsis Shoots Authors:  Jose P. Fonseca, Marcus Griffiths, Larry M. York and Kirankumar S. Mysore, date: 10/05/2021, view: 644, Q&A: 0

Dark respiration refers to experimental measures of leaf respiration in the absence of light, done to distinguish it from the photorespiration that occurs during photosynthesis. Dark aerobic respiration reactions occur solely in the mitochondria and convert glucose molecules from cytoplasmatic glycolysis and oxygen into carbon dioxide and water,

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