• Volume 12, 2022
  • 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.4344

Past Issues in 2022

Volume: 12 Issue: 5

Biochemistry

Rapid Lipid Quantification in Caenorhabditis elegans by Oil Red O and Nile Red Staining Authors:  Nicole L. Stuhr, James D. Nhan, Amy M. Hammerquist, Bennett Van Camp, David Reoyo and Sean P. Curran, date: 03/05/2022, view: 2147, Q&A: 0

The ability to stain lipid stores in vivo allows for the facile assessment of metabolic status in individuals of a population following genetic and environmental manipulation or pharmacological treatment. In the animal model Caenorhabditis elegans, lipids are stored in and mobilized from intracellular lipid droplets in the intestinal and

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Biological Engineering

Laser Microirradiation and Real-time Recruitment Assays Using an Engineered Biosensor Authors:  Carolina dos Santos Passos, Robert E. Cohen and Tingting Yao, date: 03/05/2022, view: 1526, Q&A: 0

Double-strand breaks (DSBs) are lesions in DNA that, if not properly repaired, can cause genomic instability, oncogenesis, and cell death. Multiple chromatin posttranslational modifications (PTMs) play a role in the DNA damage response to DSBs. Among these, RNF168-mediated ubiquitination of lysines 13 or 15 at the N-terminal tail of histone H2A

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Cancer Biology

Whole-genome Methylation Analysis of APOBEC Enzyme-converted DNA (~5 kb) by Nanopore Sequencing Authors:  Suzuko Zaha, Yoshitaka Sakamoto, Satoi Nagasawa, Sumio Sugano, Ayako Suzuki, Yutaka Suzuki and Masahide Seki, date: 03/05/2022, view: 1748, Q&A: 0

In recent years, DNA methylation research has been accelerated by the advent of nanopore sequencers. However, read length has been limited by the constraints of base conversion using the bisulfite method, making analysis of chromatin content difficult. The read length of the previous method combining bisulfite conversion and long-read sequencing

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Cell Biology

Colocalization Analysis for Cryosectioned and Immunostained Tissue Samples with or without Label Retention Expansion Microscopy (LR-ExM) by JACoP Authors:  Xiang Zhao, Jason Garcia, Loic A. Royer and Su Guo, date: 03/05/2022, view: 1788, Q&A: 0

Asymmetric cell division (ACD) is fundamental for balancing cell proliferation and differentiation in metazoans. During active neurogenesis in the developing zebrafish forebrain, radial glia progenitors (RGPs) mainly undergo ACD to produce one daughter with high activity of Delta/Notch signaling (proliferative cell fate) and another daughter with

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A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow Authors:  Lei Dang, Nanxi Li, Xiaohao Wu, Dijie Li, Zongkang Zhang, Bao-Ting Zhang, Aiping Lyu, Lin Chen, Ge Zhang and Jin Liu, date: 03/05/2022, view: 1236, Q&A: 0

Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for

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Labeling Endogenous Proteins Using CRISPR-mediated Insertion of Exon (CRISPIE) Authors:  Evan A. Wilson, Tianyi Mao and Haining Zhong, date: 03/05/2022, view: 1978, Q&A: 0

The CRISPR/Cas9 technology has transformed our ability to edit eukaryotic genomes. Despite this breakthrough, it remains challenging to precisely knock-in large DNA sequences, such as those encoding a fluorescent protein, for labeling or modifying a target protein in post-mitotic cells. Previous efforts focusing on sequence insertion to the

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Developmental Biology

Labeling and Tracking Mitochondria with Photoactivation in Drosophila Embryos Authors:  Sayali Chowdhary and Richa Rikhy, date: 03/05/2022, view: 1302, Q&A: 0

Mitochondria are relatively small, fragmented, and abundant in the large embryos of Drosophila, Xenopus and zebrafish. It is essential to study their distribution and dynamics in these embryos to understand the mechanistic role of mitochondrial function in early morphogenesis events. Photoactivation of mitochondrially tagged GFP (mito-PA-GFP) is

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Microbiology

High-speed Atomic Force Microscopy Observation of Internal Structure Movements in Living Mycoplasma Authors:  Kohei Kobayashi, Noriyuki Kodera and Makoto Miyata, date: 03/05/2022, view: 1354, Q&A: 0

Dozens of Mycoplasma species belonging to the class Mollicutes bind to solid surfaces through the organelle formed at a cell pole and glide in its direction by a unique mechanism. In Mycoplasma mobile, the fastest gliding species in Mycoplasma, the force for gliding is generated by ATP hydrolysis on an internal structure. However, the spatial and

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RNA-mediated in vivo Directed Evolution in Yeast Authors:  Emil D. Jensen and Michael K. Jensen, date: 03/05/2022, view: 1323, Q&A: 0

Directed evolution is a powerful approach to obtain genetically-encoded sought-for traits. Compared to the prolonged adaptation regimes to mutations occurring under natural selection, directed evolution unlocks rapid screening and selection of mutants with improved traits from vast mutated sequence spaces. Many systems have been developed to

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A Novel PCR-Based Methodology for Viral Detection Utilizing Mechanical Homogenization Authors:  Zachary P. Morehouse, Caleb M. Proctor, Gabriella L. Ryan and Rodney J. Nash, date: 03/05/2022, view: 721, Q&A: 0

The impact of viral diseases on human health is becoming increasingly prevalent globally with the burden of disease being shared between resource-rich and poor areas. As seen in the global pandemic caused by SARS-CoV-2, there is a need to establish viral detection techniques applicable to resource-limited areas that provide sensitive and specific

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Neuroscience

Optogenetic Targeting of Mouse Vagal Afferents Using an Organ-specific, Scalable, Wireless Optoelectronic Device Authors:  Sungcheol Hong, Woo Seok Kim, Yong Han, Rahul Cherukuri, Haemin Jung, Carlos Campos, Qi Wu and Sung Il Park, date: 03/05/2022, view: 699, Q&A: 0

Optogenetics has the potential to transform the study of the peripheral nervous system (PNS), but the complex anatomy of the PNS poses unique challenges for the focused delivery of light to specific tissues. This protocol describes the fabrication of a wireless telemetry system for studying peripheral sensory pathways. Unlike existing wireless

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Operant Self-medication for Assessment of Spontaneous Pain Relief and Drug Abuse Liability in Mouse Models of Chronic Pain Authors:  David Cabañero, Beltrán Álvarez-Pérez, Elena Martín-García and Rafael Maldonado, date: 03/05/2022, view: 527, Q&A: 0

The search for safe and efficient chronic pain treatments is dampened by the lack of reliable models that faithfully reproduce current pharmacological treatments for chronic spontaneous pain in humans. Preclinical models often assess the antinociceptive efficacy of non-contingent pharmacological treatments evaluated in the short-term. Here, we

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A Novel Standardized Peripheral Nerve Transection Method and a Novel Digital Pressure Sensor Device Construction for Peripheral Nerve Crush Injury Authors:  Jung Il Lee, Grant D. Wandling, M A Hassan Talukder, Prem Kumar Govindappa and John C. Elfar, date: 03/05/2022, view: 538, Q&A: 0

Peripheral nerve injury (PNI) is common in all walks of life, and the most common PNIs are nerve crush and nerve transection. While optimal functional recovery after crush injury occurs over weeks, functional recovery after nerve transection with microsurgical repair and grafting is poor, and associated with permanent disability. The gold-standard

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Plant Science

A Quick Method to Quantify Iron in Arabidopsis Seedlings Authors:  Chandan Kumar Gautam, Huei-Hsuan Tsai and Wolfgang Schmidt, date: 03/05/2022, view: 1110, Q&A: 0

Iron (Fe) is an indispensable micronutrient for plant growth and development. Since both deficiency, as well as a surplus of Fe, can be detrimental to plant health, plants need to constantly tune uptake rates to maintain an optimum level of Fe. Quantification of Fe serves as an important parameter for analyzing the fitness of plants from different

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Stem Cell

Skeletal Stem Cell Isolation from Cranial Suture Mesenchyme and Maintenance of Stemness in Culture Authors:  Takamitsu Maruyama, Hsiao-Man Ivy Yu and Wei Hsu, date: 03/05/2022, view: 578, Q&A: 0

Skeletal stem cells residing in the suture mesenchyme are responsible for calvarial development, homeostatic maintenance, and injury-induced repair. These naïve cells exhibit long-term self-renewal, clonal expansion, and multipotency. They possess osteogenic abilities to regenerate bones in a cell-autonomous manner and can directly replace

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