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Generation of Organotypic Raft Cultures from Primary Human Keratinocytes
An in vitro method to mimic in vivo epithelial differentiation is described. Many viruses target epithelial cells as part of their viral life cycle, and this method provides a means of examining virus:host interactions that more closely resembles that which occurs in vivo. This technique can be used with primary keratinocytes, established cell lines, as well as normal or diseased biopsy tissue.
Launch Video
Published: 02/22/2012 ID: 3668
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Daniel Anacker1 Cary Moody1, 2
1Department of Microbiology & Immunology, University of North Carolina-Chapel Hill 2Lineberger Cancer Center, University of North Carolina-Chapel Hill |
Immunology, Issue 60, Epithelium, organotypic raft culture, virus, keratinocytes, papillomavirus, |
Reproducible Mouse Sciatic Nerve Crush and Subsequent Assessment of Regeneration by Whole Mount Muscle Analysis
In this report we describe a method to crush mouse sciatic nerve. This method uses readily available hemostatic forceps and easily and reproducibly produces complete sciatic nerve crush. In addition, we describe a method to prepare muscle whole mounts suitable for analysis of nerve regeneration after sciatic nerve crush.
Launch Video
Published: 02/22/2012 ID: 3606
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Andrew R. Bauder Toby A. Ferguson
Center for Neural Repair and Rehabilitation, Temple University |
Neuroscience, Issue 60, Sciatic nerve crush, regeneration, neuromuscular junction, muscle whole mount, mouse, |
Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks
The collection of infected tick hemolymph, salivary glands, and saliva is important to study how tick-borne pathogens cause disease. In this protocol we demonstrate how to collect hemolymph and salivary glands from feeding Ixodes scapularis nymphs. We also demonstrate saliva collection from female I. scapularis adults.
Launch Video
Published: 02/21/2012 ID: 3894
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Toni G. Patton1 Gabrielle Dietrich2 Kevin Brandt1 Marc C. Dolan2 Joseph Piesman2 Robert D. Gilmore Jr.1
1Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention 2Tick-Borne Diseases Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention |
Immunology, Issue 60, Ixodes scapularis, Lyme disease, Borrelia burgdorferi, salivary glands, hemolymph, tick dissection, saliva, tick, |
Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence
Here we describe a method to directly visualize microregional tissue hypoxia in the mouse cortex in vivo. It is based on concurrent two-photon imaging of nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation. This method is useful for high resolution analysis of tissue oxygen supply.
Launch Video
Published: 02/21/2012 ID: 3466
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Oksana Polesskaya1 Anita Sun2 Gheorghe Salahura2 Jharon N. Silva1 Stephen Dewhurst1 Karl Kasischke3
1Department of Microbiology and Immunology, University of Rochester Medical Center 2Center for Neural Development and Disease, University of Rochester Medical Center 3Deptartment of Neurology, Center for Neural Development and Disease, University of Rochester Medical Center |
Neuroscience, Issue 60, mouse, two-photon, cortex, nicotinamide adenine dinucleotide, angiography, hypoxia, |
Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts
We present a protocol for efficient reprogramming of human somatic cells into human induced pluripotent stem cells (hiPSC) using retroviral vectors encoding Oct3/4, Sox2, Klf4 and c-myc (OSKM) and identification of correctly reprogrammed hiPSC by live staining with Tra-1-81 antibody.
Launch Video
Published: 02/20/2012 ID: 3416
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Urszula Polak1, 2 Calley Hirsch1 Sherman Ku3 Joel Gottesfeld3 Sharon Y.R. Dent1 Marek Napierala1
1Department of Molecular Carcinogenesis and Center for Cancer Epigenetics, University of Texas M.D. Anderson Cancer Center 2Department of Cell Biology, Poznan University of Medical Sciences 3Department of Molecular Biology, The Scripps Research Institute |
Developmental Biology, Issue 60, stem cells, induced pluripotent stem cells, iPSC, somatic cell reprogramming, pluripotency, retroviral transduction, |
A PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica
We provide a cost-effective and rapid molecular genotyping protocol that employs variety-specific PCR primers that target DNA sequence differences within the chloroplast trnL-F spacer region to differentiate between varieties of Imperata cylindrica (cogongrass) that cannot be distinguished by morphology alone. These varieties include the federally listed noxious weed, cogongrass and closely-related, wide-spread ornamental variety, I. cylindrica var. koenigii (Japanese blood grass).
Launch Video
Published: 02/20/2012 ID: 3265
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Leland J. Cseke1 Sharon M. Talley2
1Department of Biological Sciences, The University of Alabama, Huntsville 2USDA-APHIS-PPQ, Center for Plant Health Science and Technology |
Molecular Biology, Issue 60, Molecular genotyping, Japanese blood grass, Red Baron, cogongrass, invasive plants, |
Morphometric Analyses of Retinal Sections
This video demonstrates three types of morphometric analyses of the retina, which include measuring the inner nuclear layer thickness, quantifying the number of retinal ganglion cells (RGCs) and measuring the sizes of RGCs. The technique can offer a simple but scientific platform for morphometric analyses.
Launch Video
Published: 02/19/2012 ID: 3377
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Tin Fung Chan1 Kin Chiu1 Carmen Ka Ming Lok1 Wing Lau Ho1 Kwok-Fai So1, 2, 3 Raymond Chuen-Chung Chang1, 2, 3
1Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong 2Research Centre of Heart, Brain, Hormone and Healthy Aging, LKS Faculty of Medicine, The University of Hong Kong 3State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong |
Neuroscience, Issue 60, morphometric analysis, retina, thickness, cell size, Stereo Investigator, neuroscience, |
Preterm EEG: A Multimodal Neurophysiological Protocol
This video explains the background theory of the neonatal EEG activity and the sensory responses, followed by a live demonstration of their recording in neonatal intensive care unit.
Launch Video
Published: 02/18/2012 ID: 3774
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Susanna Stjerna1 Juha Voipio2 Marjo Metsäranta3 Kai Kaila2, 4 Sampsa Vanhatalo1
1Department of Children's Clinical Neurophysiology, Helsinki University Hospital, University of Helsinki 2Department of Biosciences, University of Helsinki 3Department of Pediatrics, Helsinki University Hospital, University of Helsinki 4Neuroscience Center, University of Helsinki |
Neuroscience, Issue 60, neurophysiology, preterm baby, neonatal, EEG, evoked response, high density EEG, FbEEG, sensory evoked response, neonatal intensive care unit, |
In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis
The mesothelial clearance assay described here takes advantage of fluorescently labeled cells and time-lapse video microscopy to visualize and quantitatively measure the interactions of ovarian cancer multicellular spheroids and mesothelial cell monolayers. This assay models the early steps of ovarian cancer metastasis.
Launch Video
Published: 02/17/2012 ID: 3888
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Rachel A. Davidowitz Marcin P. Iwanicki Joan S. Brugge
Department of Cell Biology, Harvard Medical School |
Medicine, Issue 60, Ovarian Cancer, Metastasis, In vitro Model, Mesothelial, Spheroid, |
MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression
We have developed 3D coculture models for live-cell imaging in real-time of interactions among breast tumor cells and other cells in their microenvironment that impact progression to an invasive phenotype. These models can serve as preclinical screens for drugs to target paracrine-induced proteolytic, chemokine/cytokine and kinase pathways implicated in invasiveness.
Launch Video
Published: 02/17/2012 ID: 3661
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Mansoureh Sameni1 Arulselvi Anbalagan1 Mary B. Olive1 Kamiar Moin1, 2 Raymond R. Mattingly1, 2 Bonnie F. Sloane1, 2
1Department of Pharmacology, Wayne State University 2Barbara Ann Karmanos Cancer Institute, Wayne State University |
Medicine, Issue 60, Immunology, Breast, cancer, extracellular matrix, invasion, proteolysis, tumor microenvironment, |