Isolation and Characterization of Dendritic Cells and…
Published 5/21/2012
Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition
1Graduate Group in Biophysics, University of California San Francisco , 2Department of Biochemistry and Biophysics, University of California San Francisco
Visualizing protein samples by negative stain electron microscopy (EM) has become a popular structural analysis method. It is useful for quantitative structural analysis, such as calculating a 3D reconstruction of the molecules being studied, and also for qualitative examination of the quality of protein preparations. In this article we present detailed protocols for preparing the EM grids, staining the sample and visualizing the sample in an electron microscope. Novice users can follow these protocols easily and to utilize negative stain EM as a routine assay, in addition to other biochemical assays, for evaluating their protein samples.
Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering
Biomedical Engineering Department, Worcester Polytechnic Institute
This article outlines a versatile method to create cell-derived tissue rings by cellular self-assembly. Smooth muscle cells seeded into ring-shaped agarose wells aggregate and contract to form robust three-dimensional (3D) tissues within 7 days. Millimeter-scale tissue rings are conducive to mechanical testing and serve as building blocks for tissue assembly.
Knowing What Counts: Unbiased Stereology in the Non-human Primate Brain
1Department of Physiology, University of Montreal, 2Ecole d’optometrie, University of Montreal, 3Stereology Resource Center
The anatomical organization of the primate brain can provide important insights into normal and pathological conditions in humans. Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given reference space1.
Bioelectric Analyses of an Osseointegrated Intelligent Implant Design System for Amputees
1Department of Veteran Affairs, 2Department of Bioengineering, University of Utah, 3Scientific Computing and Imaging Institute , University of Utah, 4Department of Physical Medicine and Rehabilitation, University of Utah, 5Department of Orthopaedics, University of Utah
There is a need to develop alternative prosthesis attachment due to limb loss attributed to vascular occlusive diseases and trauma. The goal of the work is to introduce an osseointegrated intelligent implant design system to increase skeletal fixation and reduce periprosthetic infection rates for patients needing osseointegrated technology.
Ex vivo Mechanical Loading of Tendon
1Department of Bioengineering, University of California, Berkeley , 2Department of Bioengineering, University of California, Berkeley; Division of Occupational Medicine, University of California, San Francisco
A new in vitro system for simultaneously loading four tendons in culture is described.
Cecal Ligation Puncture Procedure
1Department of Microbiology and Immunology School of Medicine, Temple University , 2Department of Biochemistry, School of Medicine, Temple University
The mouse model of cecal ligation and puncture as a valuable tool for the study of human sepsis.
Organotypic Culture of Full-thickness Adult Porcine Retina
1Neurology and Neurosciences, University of Medicine and Dentistry of New Jersey - UMDNJ, 2Institute of Ophthalmology and Visual Science, University of Medicine and Dentistry of New Jersey - UMDNJ
Here we describe a cost-effective technique for organotypic culture of adult porcine retina for seven days. Briefly, a sterile filter paper was used to lift the neural retina off from the RPE and place photoreceptor side up on an insert raised by a custom-made stand.
Ex Vivo Culture of Patient Tissue & Examination of Gene Delivery
1Cork Cancer Research Centre, Mercy University Hospital and Leslie C. Quick Jnr. Laboratory, University College Cork, 2Department of Computer Science, University College Cork
This article describes the culture of patient tissue slices for gene delivery studies and subsequent analysis of gene expression using IVIS bioluminescence imaging.
How to Measure Cortical Folding from MR Images: a Step-by-Step Tutorial to Compute Local Gyrification Index
1Department of Psychiatry, University of Geneva School of Medicine, 2Signal Processing Laboratory, École Polytechnique Fédérale de Lausanne, 3Department of Radiology, University Hospital Center and University of Lausanne, 4Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital
Measuring gyrification (cortical folding) at any age represents a window into early brain development. Hence, we previously developed an algorithm to measure local gyrification at thousands of points over the hemisphere1. In this paper, we detail the computation of this local gyrification index.
An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain
Department of Molecular Biomedical Sciences, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University
This protocol describes an organotypic slice assay optimized for the postnatal brain and high-resolution time-lapse imaging of neuroblast migration in the rostral migratory stream.
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