JoVE Biology welcomes all general biology research methodologies. Content in this section canvases all fields of cell, molecular, and organismal biology, ranging from new applications of standard techniques to novel approaches aimed at understanding the functions of life and living organisms. This diverse section includes, but is not limited to, techniques in physical biology, cellular biochemistry, genetics, physiology, systems biology and a combination of eukaryotic and prokaryotic model systems.
1Tactical Combat Casualty Care Research, U.S. Army Institute of Surgical Research, 2Department of Pharmacology and Toxicology, Michigan State University, 3Southwest National Primate Research Center, Texas Biomedical Research Institute
The purpose of this protocol is to demonstrate the principles and techniques for measuring and calculating glomerular filtration rate, urine flow rate, and excretion of sodium and potassium in a rat. This demonstration can be used to provide students with an overall conceptual understanding of how to measure renal function.
Published July 26, 2015. Keywords: Physiology, kidney, rat, glomerular filtration rate, urine flow rate, sodium excretion, potassium excretion, filtered load, blood pressure
1Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 2Proteomics Platform, Broad Institute
This protocol describes a procedure for enriching ECM proteins from tissues or tumors and deglycosylating and digesting the ECM-enriched preparations into peptides to analyze their protein composition by mass spectrometry.
Published July 23, 2015. Keywords: Biochemistry, Extracellular Matrix, Matrisome, Proteins, Decellularization, Sequential solubilization, Deglycosylation, Tryptic digestion, Proteomics, Mass Spectrometry
1Department of Medicine, Duke University Medical Center, 2Department of Chemistry, University of North Carolina - Chapel Hill, 3Renovo Neural Incorporated
Here, we introduce a method, cocem3D, to unveil the ultrastructure of a specific cell in its native tissue by bridging confocal and serial block-face scanning electron microscopy.
Published July 19, 2015. Keywords: Cellular Biology, Gut-Brain neurobiology, 3D electron microscopy, serial block-face scanning electron microscopy, chemosensory cell, neuropod, nutrient-sensing, gastrointestinal chemosensation
1Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia
Here we describe a simple method for the isolation of spatially distinct murine colonic epithelial surface cells from the underlying crypt-base cells.
Published July 12, 2015. Keywords: Cellular Biology, epithelia, mucosa, colon, crypt, microdissection, microtome
1Department of Molecular and Cell Biology, Center for Systems Biology, University of Texas at Dallas
Altered intracellular heme levels are associated with common diseases such as cancer. Thus, there is a need to measure heme biosynthesis levels in diverse cells. The goal of this protocol is to provide a fast and sensitive method to measure and compare the levels of heme synthesis in different cells.
Published July 9, 2015. Keywords: Molecular Biology, Heme, heme synthesis level, mammalian cells, [4-14C] 5-aminolevulinic acid ([14C] 5-ALA), cancer
1Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern, 2Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, 3Department of Oral and Maxillofacial Surgery, School of Dental Medicine, Universitat Internacional de Catalunya, 4Robert K. Schenk Laboratory of Oral Histology, School of Dental Medicine, University of Bern, 5Department of Cranio Maxillofacial Surgery, Inselspital, University of Bern, 6Department of Implant Dentistry, School of Dentistry, Universidade Federal de Santa Catarina
We describe here how to prepare bone-conditioned medium (BCM) and test its activity in vitro.
Published July 8, 2015. Keywords: Molecular Biology, Bone Conditioned Medium, BCM, bone autograft, guided bone regeneration, GBR, dental implant, membrane, supernatant, growth factors, contour augmentation, autologous bone
1Radiological Protection Research and Instrumentation, Canadian Nuclear Laboratories
A protocol to evaluate changes in DNA damage levels and DNA repair capacity that may be induced by chronic in vivo low dose irradiation in mouse spleen lymphocytes, by measuring phosphorylated histone H2AX, a marker of DNA double-strand breaks, using flow cytometry is presented.
Published July 3, 2015. Keywords: Molecular Biology, DNA damage, DNA double-strand breaks, DNA repair, γ-H2AX, low dose radiation, tritium, β--radiation, γ-radiation, chronic exposure, flow cytometry, in vivo, mouse model
1Department of Anesthesiology and Perioperative Care, University of California Irvine, 2Department of Biomedical and Molecular Sciences, Queen's University, 3Department of Pharmacology, University of California Irvine
Receptor trafficking modulates signaling and cell responsiveness to ligands and is, itself, responsive to cell conditions, including ligand-induced signaling. Here, we describe a powerful and flexible technique for quantitatively assessing drug-induced receptor trafficking using immunolabeling and colocalizational analysis.
Published July 3, 2015. Keywords: Molecular Biology, Trafficking, Receptor, G-protein coupled receptor, Internalization, Degradation, Recycling, Colocalization, Immunocytochemistry, Immunohistochemistry, Microscopy
1Department of Physiology, Medical College of Wisconsin
Changes in the intracellular calcium levels in the podocytes are one of the most important means to control the filtration function of glomeruli. Here we explain a high-throughput approach that allows detection of real-time calcium handling and single ion channels activity in the podocytes of the freshly isolated glomeruli.
Published June 27, 2015. Keywords: Molecular Biology, glomeruli, podocytes, calcium imaging, ion channels, confocal microscopy, intracellular calcium.
1Biological Effects Department, Centre for Radiation, Chemicals and Environmental Hazards, Public Health England
We report an in vitro method that allows the quantitation of the actual number of adhesive cells within an endothelial cell monolayer.
Published June 18, 2015. Keywords: Cellular Biology, Atherosclerosis, Endothelial cells, Monocytes, Adhesion, Inflammation, Adhesion assay