• 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.4231

Current Issue in 2021

Volume: 11 Issue: 22

Biophysics

Cortical Laminar Recording of Multi-unit Response to Distal Forelimb Electrical Stimulation in Rats Authors:  Charles-Francois V. Latchoumane, Rameen Forghani and Lohitash Karumbaiah, date: 11/20/2021, view: 835, Q&A: 0

Severe traumatic brain injury (sTBI) survivors experience permanent functional disabilities due to significant volume loss and the brain’s poor capacity to regenerate. Chondroitin sulfate glycosaminoglycans (CS-GAGs) are key regulators of growth factor signaling and neural stem cell homeostasis in the brain. In this protocol, we describe how

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Macroscopic Structural and Connectome Mapping of the Mouse Brain Using Diffusion Magnetic Resonance Imaging Authors:  Tanzil Mahmud Arefin, Choong Heon Lee, Jordon D. White, Jiangyang Zhang and Arie Kaffman, date: 11/20/2021, view: 774, Q&A: 0

Translational work in rodents elucidates basic mechanisms that drive complex behaviors relevant to psychiatric and neurological conditions. Nonetheless, numerous promising studies in rodents later fail in clinical trials, highlighting the need for improving the translational utility of preclinical studies in rodents. Imaging of small rodents

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Cancer Biology

Measurement of DNA Damage Using the Neutral Comet Assay in Cultured Cells Authors:  Elena Clementi, Zuzana Garajova and Enni Markkanen, date: 11/20/2021, view: 1015, Q&A: 0

Maintenance of DNA integrity is of pivotal importance for cells to circumvent detrimental processes that can ultimately lead to the development of various diseases. In the face of a plethora of endogenous and exogenous DNA damaging agents, cells have evolved a variety of DNA repair mechanisms that are responsible for safeguarding genetic

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Measurement of Bone Metastatic Tumor Growth by a Tibial Tumorigenesis Assay Authors:  Baotong Zhang, Xin Li, Wei-Ping Qian, Daqing Wu and Jin-Tang Dong, date: 11/20/2021, view: 699, Q&A: 0

Bone metastasis is a frequent and lethal complication of many cancer types (i.e., prostate cancer, breast cancer, and multiple myeloma), and a cure for bone metastasis remains elusive. To recapitulate the process of bone metastasis and understand how cancer cells metastasize to bone, intracardiac injection and intracaudal arterial animal models

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Cell Biology

Pericyte Mapping in Cerebral Slices with the Far-red Fluorophore TO-PRO-3 Authors:  Sandra P. Mai-Morente, Juan P. Irigoyen, Victoria M. Carriquiry, Virginia M. Marset, Mariana Di Doménico, Eugenia Isasi and Verónica Abudara, date: 11/20/2021, view: 746, Q&A: 0

This protocol describes a method for high-resolution confocal imaging of pericytes with the far-red fluorophore TO-PROTM-3 Iodide 642/661 in cerebral slices of murine. Identification of pericytes with TO-PRO-3 is a short time-consuming, high cost-effective and robust technique to label pericytes with no need for immunostaining or generation of

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Live Imaging of Apoptotic Extrusion and Quantification of Apical Extrusion in Epithelial Cells Authors:  Kinga Duszyc, Ivar Noordstra, Alpha S. Yap and Guillermo A. Gomez, date: 11/20/2021, view: 765, Q&A: 0

Apoptotic cell death eliminates unhealthy cells and maintains homeostatic cell numbers within tissues. Epithelia, which serve as fundamental tissue barriers for the body, depend on a physical expulsion of dying cells (apoptotic cell extrusion) to remain sealed and intact. Apoptotic cell extrusion has been widely studied over recent years, with

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Gamete Fusion Assay in Mice Author:  Naokazu Inoue, date: 11/20/2021, view: 285, Q&A: 0

Gamete fusion, which is the final event of fertilization, is a crucial physiological event in the creation of a new fetus. In mammals, sperm IZUMO1 and oocyte IZUMO1R (JUNO) recognition play a role in triggering this process. Gamete fusion occurs through a complex but steady and unfailing intermolecular reaction because fertilization must ensure

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Developmental Biology

Isolation of Primary Leydig Cells from Murine Testis Authors:  Jinlian Liang, Yan Tang, Hanhao Li, Jiaxin Mei, Zhen Cao, Huan Xia, Rufei Huang, Yan Yang and Yadong Huang, date: 11/20/2021, view: 302, Q&A: 0

In males, Leydig cells are the primary source of testosterone, which is necessary for testis development, masculinization, and spermatogenesis. Leydig cells are a valuable cellular model for basic research; thus, it is important to develop an improved method for isolation and purification of Leydig cells from testes. The available methods for

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Wholemount in situ Hybridization for Spatial-temporal Visualization of Gene Expression in Early Post-implantation Mouse Embryos Authors:  Xianfa Yang, Yingying Chen, Lu Song, Ting Zhang and Naihe Jing, date: 11/20/2021, view: 344, Q&A: 0

Regionalized distribution of genes plays crucial roles in the formation of the spatial pattern in tissues and embryos during development. In situ hybridization has been one of the most widely used methods to screen, identify, and validate the spatial distribution of genes in tissues and embryos, due to its relative simplicity and low cost.

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Immunology

CD45 Immunohistochemistry in Mouse Kidney Authors:  Shirong Zheng and Paul N. Epstein, date: 11/20/2021, view: 382, Q&A: 0

CD45 is a pan-leukocyte marker, and CD45 stain is widely used to determine the extent of inflammatory cell infiltration and its association with tissue injury. In this manuscript, we share a reliable immunohistochemistry (IHC) protocol for CD45 staining in sections of paraffin-embedded mouse kidney. A rat anti-CD45 antibody was used as primary

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Microbiology

Analysis of Protein Stability by Synthesis Shutoff Authors:  Ira Buntenbroich, Tânia Simões and Mafalda Escobar-Henriques, date: 11/20/2021, view: 519, Q&A: 0

In this protocol, we describe the analysis of protein stability over time, using synthesis shutoff. As an example, we express HA-tagged yeast mitofusin Fzo1 in Saccharomyces cerevisiae and inhibit translation via cycloheximide (CHX). Proteasomal inhibition with MG132 is performed, as an optional step, before the addition of CHX. Proteins are

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Neuroscience

Knockoff: Druggable Cleavage of Membrane Proteins Authors:  John H. Rinald, Jason D. Vevea and Edwin R. Chapman, date: 11/20/2021, view: 477, Q&A: 0

Comparative cell biology relies on methods that disrupt protein function. Traditional approaches target the gene that encodes the protein of interest via conventional knockout (KO) methodology, conditional Cre-lox system, or recently, flexible protocols based on CRISPR/Cas9. However, these technologies lack precise temporal control (hours),

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Visual-stimuli Four-arm Maze test to Assess Cognition and Vision in Mice Authors:  Jean-Philippe Vit, Dieu-Trang Fuchs, Ariel Angel, Aharon Levy, Itschak Lamensdorf, Keith L. Black, Yosef Koronyo and Maya Koronyo-Hamaoui, date: 11/20/2021, view: 337, Q&A: 0

Visual impairments, notably loss of contrast sensitivity and color vision, were documented in Alzheimer’s disease (AD) patients yet are critically understudied. This protocol describes a novel visual-stimuli four-arm maze (ViS4M; also called visual x-maze), which is a versatile x-shaped maze equipped with spectrum- and intensity-controlled

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Plant Science

Preparation and Transfection of Populus tomentosa Mesophyll Protoplasts Authors:  Hou-Ling Wang, Ting Wang, Qi Yang, Weilun Yin, Xinli Xia, Hongwei Guo and Zhonghai Li, date: 11/20/2021, view: 630, Q&A: 0

Mesophyll protoplasts freshly isolated from leaves are a useful research system in plants. However, cell walls in woody plants contain more pectin, making mesophyll protoplasts isolation difficult in Populus. This has limited their application in biochemical, molecular, cellular, genetic, genomic, transcriptomic, and proteomic assays. In this

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Imaging of Lipid Uptake in Arabidopsis Seedlings Utilizing Fluorescent Lipids and Confocal Microscopy Authors:  Rosa L. López-Marqués and Thomas G. Pomorski, date: 11/20/2021, view: 763, Q&A: 0

Eukaryotic cells use a diverse set of transporters to control the movement of lipids across their plasma membrane, which drastically affects membrane properties. Various tools and techniques to analyze the activity of these transporters have been developed. Among them, assays based on fluorescent phospholipid probes are particularly suitable,

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Stem Cell

Isolation of CD31+ Bone Marrow Endothelial Cells (BMECs) from Mice Authors:  Alhaji Osman Smith, Seyram Yao Adzraku, Wen Ju, Jianlin Qiao, Kailin Xu and Lingyu Zeng, date: 11/20/2021, view: 387, Q&A: 0

In the bone marrow microenvironment, endothelial cells (ECs) play a pivotal role in regulating the production of both growth and inhibiting factors. They are held together by adherence molecules that interact with hematopoietic progenitor cells. The study of ECs in the hematopoietic stem cell niche is limited due to the lack of efficient protocols

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