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Editors' Choice
Editors' Choice corresponds to recently published bio-protocols (past one year) that chosen by Bio-protocol Associate Editors based on the protocols' quality, novelty, or potential impact.
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Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
Authors:  Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya, date: 01/20/2018, view: 330, Q&A: 0
Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation of equivalent cells from pluripotent stem cells has been limited. Using a cocktail of small molecules that we previously found useful in stable culture ...
Fluorescent Measurement of Synaptic Activity Using FM Dyes in Dissociated Hippocampal Cultured Neurons
Authors:  Roman M. Lazarenko, Claire E. DelBove and Qi Zhang, date: 01/20/2018, view: 218, Q&A: 0
Release and recycling of synaptic vesicles are essential for neurotransmission and synaptic plasticity. To gain mechanistic understanding of these processes, direct measurements of vesicle release and retrieval is indispensable. Styryl dyes like FM1-43 and FM4-64 have been widely used for this purpose and their loading and unloading are reliable ...
Multiple Stepwise Gene Knockout Using CRISPR/Cas9 in Escherichia coli
Authors:  Enrico König, Francesca Zerbini, Ilaria Zanella, Davide Fraccascia and Guido Grandi, date: 01/20/2018, view: 678, Q&A: 0
With the recent implementation of the CRISPR/Cas9 technology as a standard tool for genome editing, laboratories all over the world are undergoing one of the biggest advancements in molecular biology since PCR. The key advantage of this method is its simplicity and universal applicability for species of any phylum. Of particular interest is the ...
Micro-computed Tomography to Visualize Vascular Networks in Maize Stems
Authors:  Akiteru Maeno and Katsutoshi Tsuda, date: 01/05/2018, view: 340, Q&A: 0
Plant vascular systems in the stem connect roots with aerial organs to move solutes containing minerals, nutrients as well as signaling molecules, and therefore, they play pivotal roles in plant growth and development. However, stem vascular systems, especially in crop species, have been poorly described since they are deeply embedded in the ...
Analysing Temporal Dynamics of T Cell Division in vivo Using Ki67 and BrdU Co-labelling by Flow Cytometry
Authors:  Thea Hogan, Andrew Yates and Benedict Seddon, date: 12/20/2017, view: 502, Q&A: 0
This protocol was developed to increase the richness of information available from in vivo T cell proliferation studies. DNA labelling techniques such as BrdU incorporation allow precise control of label administration and withdrawal, so that the division history of a population can be tracked in detail over long timeframes (days-weeks). ...
Mouse Model of Immune Complex-mediated Vasculitis in Dorsal Skin and Assessment of the Neutrophil-mediated Tissue Damage
Neutrophils are the most abundant leukocytes in the blood. In the recent decades, their crucial roles in host defense, immune regulation and tissue damage have been studied in a deeper dimension. In this protocol, we described a mouse model of immune complex-mediated vasculitis in the dorsal skin induced by Arthus reaction, and the subsequent ...
Live Imaging of Axonal Transport in the Motor Neurons of Drosophila Larvae
Authors:  Tsuyoshi Inoshita, Nobutaka Hattori and Yuzuru Imai, date: 12/05/2017, view: 556, Q&A: 0
Axonal transport, which is composed of microtubules, motor proteins and a variety of types of cargo, is a prominent feature of neurons. Monitoring these molecular dynamics is important to understand the biological processes of neurons as well as neurodegenerative disorders that are associated with axonal dysfunction. Here, we describe a protocol ...
Instillation of Particulate Suspensions to the Lungs
Authors:  Etsushi Kuroda, Yasuo Morimoto and Ken J Ishii, date: 11/20/2017, view: 793, Q&A: 0
Inhaled fine particulates are thought to cause chronic pulmonary inflammation through the deposition of particulates into the lungs. To investigate the effect of fine particulates on the lungs, instillation of suspension of particulates into the lungs is required. This protocol describes direct injection of suspensions of fine particulates into ...
Cell-free Fluorescent Intra-Golgi Retrograde Vesicle Trafficking Assay
Authors:  Nathanael P. Cottam and Daniel Ungar, date: 11/20/2017, view: 710, Q&A: 0
Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was ...
Rapid IFM Dissection for Visualizing Fluorescently Tagged Sarcomeric Proteins
Authors:  Yu Shu Xiao, Frieder Schöck and Nicanor González-Morales, date: 11/20/2017, view: 702, Q&A: 0
Sarcomeres, the smallest contractile unit of muscles, are arguably the most impressive actomyosin structure. Yet a complete understanding of sarcomere formation and maintenance is missing. The Drosophila indirect flight muscle (IFM) has proven to be a very valuable model to study sarcomeres. Here, we present a protocol for the rapid ...