• 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.4049

Past Issues in 2021

Volume: 11 Issue: 11

Biophysics

Preparation of Doublet Microtubule Fraction for Single Particle Cryo-electron Microscopy Authors:  Corbin Black, Daniel Chen Dai, Katya Peri, Muneyoshi Ichikawa and Khanh Huy Bui, date: 06/05/2021, view: 615, Q&A: 0

Over the years, studying the ultrastructure of the eukaryotic cilia/flagella using electron microscopy (EM) has contributed significantly toward our understanding of ciliary function. Major complexes in the cilia, such as inner and outer dynein arms, radial spokes, and dynein regulatory complexes, were originally discovered by EM. Classical

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Cell Biology

Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium Authors:  Leonie Witte, Karen Linnemannstöns, Mona Honemann-Capito and Julia Christina Gross, date: 06/05/2021, view: 495, Q&A: 0

Secretory Wnt trafficking can be studied in the polarized epithelial monolayer of Drosophila wing imaginal discs (WID). In this tissue, Wg (Drosophila Wnt-I) is presented on the apical surface of its source cells before being internalized into the endosomal pathway. Long-range Wg secretion and spread depend on secondary secretion from endosomal

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Isolation of First-Trimester and Full-term Human Placental Hofbauer Cells Authors:  Anna Appios, Jake R. Thomas and Naomi McGovern, date: 06/05/2021, view: 907, Q&A: 0

The placenta is the crucial organ that regulates the health of both mother and fetus during pregnancy. The human placenta is composed of villous tree-like structures that embed into the maternal decidua. Within the stroma of the villi resides a population of fetally-derived macrophages, the Hofbauer cells (HBC). HBC are the only fetal immune cells

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Developmental Biology

Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 447, Q&A: 0

Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several aspects of C. elegans sperm quality, including number, size, rate of activation, and mitochondrial

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Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 407, Q&A: 0

Maturation of secretory granules is a crucial process that ensures the bioactivity of cargo proteins undergoing regulated secretion. In Drosophila melanogaster, the larval salivary glands produce secretory granules that are up to four-fold larger in cross-sectional area after maturation. Therefore, we developed a live imaging microscopy approach

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Immunology

Multi-color Flow Cytometry for Comprehensive Analysis of the Tumor Immune Infiltrate in a Murine Model of Breast Cancer Authors:  Ana S. Almeida, Miriam R. Fein and Mikala Egeblad, date: 06/05/2021, view: 600, Q&A: 0

Flow cytometry is a popular laser-based technology that allows the phenotypic and functional characterization of individual cells in a high-throughput manner. Here, we describe a detailed procedure for preparing a single-cell suspension from mammary tumors of the mouse mammary tumor virus-polyoma middle T (MMTV-PyMT) and analyzing these cells by

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Microbiology

Sex-specific Separation of Plasmodium falciparum Gametocyte Populations Authors:  Melanie C. Ridgway, Daniela Cihalova and Alexander G. Maier, date: 06/05/2021, view: 405, Q&A: 0

Plasmodium falciparum is a unicellular eukaryotic parasite that causes malaria in humans. The parasite is spread by Anopheles mosquitoes after ingestion of sexual stage parasites known as gametocytes. Malaria transmission depends on parasites switching from the disease-causing asexual blood forms to male and female gametocytes. The current

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Molecular Biology

Mechanical Fractionation of Cultured Neuronal Cells into Cell Body and Neurite Fractions Authors:  Ankita Arora, Raeann Goering, Hei-Yong G. Lo and J. Matthew Taliaferro, date: 06/05/2021, view: 862, Q&A: 0

Many cells contain spatially defined subcellular regions that perform specialized tasks enabled by localized proteins. The subcellular distribution of these localized proteins is often facilitated by the subcellular localization of the RNA molecules that encode them. A key question in the study of this process of RNA localization is the

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Neuroscience

In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor Authors:  Nami Kitajima, Kenji Takikawa, Hiroshi Sekiya, Daisuke Asanuma, Hirokazu Sakamoto, Shigeyuki Namiki, Masamitsu Iino and Kenzo Hirose, date: 06/05/2021, view: 536, Q&A: 0

Adenosine 5’-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms

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Objective Quantitation of Focal Sweating Areas Using a Mouse Sweat-assay Model Authors:  Choongjin Ban and Dae-Hyuk Kwoen, date: 06/05/2021, view: 256, Q&A: 0

In vivo sweat quantitation assays are required for the development of drugs for the management of focal hyperhidrosis before clinical trials; however, in vivo assays, particularly mouse models, are rare. Even in sweat assays using mice, sweating is quantitated by manually counting the number of sweating spots, which can contribute to various

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Plant Science

Rice Root Hair Phenotypes Imaged by Cryo-SEM Authors:  Haiting Yan, Yue Wang, Jingrong Zhang, Xinru Cui, Jiasong Wu, Jie Zhou, Yuan Chen, Jia Lu, Ruiyang Guo, Maggie Ou, Hongxu Lai and Zhiming Yu, date: 06/05/2021, view: 1209, Q&A: 0

Cryo-scanning electron microscopy (cryo-SEM) was first introduced for scientific use in the 1980s. Since then, cryo-SEM has become a routine technique for studying the surfaces and internal structures of biological samples with a high water content. In contrast to traditional SEM, cryo-SEM requires no sample pretreatment processes; thus, we can

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Stem Cell

Muscle Cryoinjury and Quantification of Regenerating Myofibers in Mice Authors:  Yori Endo, Mehran Karvar and Indranil Sinha, date: 06/05/2021, view: 403, Q&A: 0

Cryoinjury, or injury due to freezing, is a method of creating reproducible, local injuries in skeletal muscle. This method allows studying the regenerative response following muscle injuries in vivo, thus enabling the evaluation of local and systemic factors that influence the processes of myofiber regeneration. Cryoinjuries are applicable to the

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Generation of Mouse Pluripotent Stem Cell-derived Trunk-like Structures: An in vitro Model of Post-implantation Embryogenesis Authors:  Adriano Bolondi, Leah Haut, Seher Ipek Gassaloglu, Polly Burton, Helene Kretzmer, René Buschow, Alexander Meissner, Bernhard G. Herrmann and Jesse V. Veenvliet, date: 06/05/2021, view: 814, Q&A: 0

Post-implantation mammalian embryogenesis involves profound molecular, cellular, and morphogenetic changes. The study of these highly dynamic processes is complicated by the limited accessibility of in utero development. In recent years, several complementary in vitro systems comprising self-organized assemblies of mouse embryonic stem cells, such

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Generation and Maintenance of Homogeneous Human Midbrain Organoids Authors:  Henrik Renner, Martha Grabos, Hans R. Schöler and Jan M. Bruder, date: 06/05/2021, view: 663, Q&A: 0

Three-dimensional cell cultures (“organoids”) promise to better recapitulate native tissue physiology than traditional 2D cultures and are becoming increasingly interesting for disease modeling and compound screening efforts. While a number of protocols for the generation of neural organoids have been published, most protocols require

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Systems Biology

Detecting Differentially Methylated Promoters in Genes Related to Disease Phenotypes Using R Authors:  Jordi Martorell Marugán and Pedro Carmona-Sáez, date: 06/05/2021, view: 312, Q&A: 0

DNA methylation in gene promoters plays a major role in gene expression regulation, and alterations in methylation patterns have been associated with several diseases. In this context, different software suites and statistical methods have been proposed to analyze differentially methylated positions and regions. Among them, the novel statistical

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Simplified Epigenome Profiling Using Antibody-tethered Tagmentation Authors:  Steven Henikoff, Jorja G. Henikoff and Kami Ahmad, date: 06/05/2021, view: 503, Q&A: 0

We previously introduced Cleavage Under Targets & Tagmentation (CUT&Tag), an epigenomic profiling method in which antibody tethering of the Tn5 transposase to a chromatin epitope of interest maps specific chromatin features in small samples and single cells. With CUT&Tag, intact cells or nuclei are permeabilized, followed by successive

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