Protocols in Current Issue
    Optical Clearing and Index Matching of Tissue Samples for High-resolution Fluorescence Imaging Using SeeDB2
    Authors:  Meng-Tsen Ke and Takeshi Imai, date: 10/20/2018, view: 3939, Q&A: 1
    [Abstract] Tissue clearing techniques are useful for large-scale three-dimensional fluorescence imaging of thick tissues. However, high-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we present a water-based optical clearing and mounting media, SeeDB2, which is ...
    Image-Based Analysis of Mitochondrial Area and Counting from Adult Mouse Dopaminergic Neurites
    Authors:  Nadee Nissanka, Carlos T Moraes and Mlena Pinto, date: 08/20/2018, view: 2910, Q&A: 0
    [Abstract] Mitochondria form dynamic cytoplasmic networks which undergo morphological changes in order to adapt to cellular stresses and signals. These changes can include alterations in size and number within a given cell. Analysis of the whole network can be a useful metric to assess overall mitochondrial health, particularly in neurons, which are highly ...
    Quantitative Electron Microscopic Assay Using Random Sampling from Single Sections to Test Plastic Synaptic Changes in Hippocampus
    Authors:  G. Mark Marcello, Lilla E. Szabó, Peter Sotonyi and Bence Rácz, date: 08/05/2018, view: 2046, Q&A: 0
    [Abstract] Studies over several decades on the organization of the CA1 hippocampus–a particularly favorable model for learning, memory and certain forms of cognition–have shown that the synaptic network in this brain region is plastic (Fortin et al., 2012). Recent evidence suggests that a number of environmental and endogenous stimuli may have a ...
    Isolation and Maintenance of Striatal Neurons
    Authors:  Luana  Naia and A. Cristina Rego, date: 04/20/2018, view: 3558, Q&A: 0
    [Abstract] Primary cultures of murine striatal neurons are widely used to explore cellular mechanisms in neurobiology, including brain diseases. Here we describe a detailed and standardized protocol to dissect and culture embryonic murine striatal neurons GABA-positive/DARPP-32-positive for 12 days in vitro, when they show good neuronal cell ...
    Photothrombotic Induction of Capillary Ischemia in the Mouse Cortex during in vivo Two-Photon Imaging
    Authors:  Robert G. Underly and Andy Y. Shih, date: 07/05/2017, view: 5094, Q&A: 0
    [Abstract] Photothrombosis of blood vessels refers to the activation of a circulating photosensitive dye with a green light to induce clotting in vivo (Watson et al., 1985). Previous studies have described how a focused green laser could be used to noninvasively occlude pial arterioles and venules at the brain surface (Schaffer et al. ...
    In vitro Assay for Dendritic Spine Retraction of Hippocampal Neurons with Sparse Labeling
    Authors:  Ryohei Iwata and Takuji Iwasato, date: 09/20/2016, view: 5803, Q&A: 0
    [Abstract] Dendritic spines are the post-synaptic structures that play a central role in excitatory synaptic transmission. Developmental spinogenesis relies on a variety of stimuli such as those derived from cell-cell communication and their downstream signaling. Here, we describe an in vitro assay of dendritic spine retraction using hippocampal ...
    Olfactory Bulb (OB) Transplants
    Authors:  Maria Savvaki and Domna Karagogeos, date: 09/05/2016, view: 4767, Q&A: 0
    [Abstract] Transplantation in mouse brain slices is a powerful tool in order to study axon targeting and migrational events during development. Taking advantage of donors and recipients belonging to different genotypes, this technique allows researchers to assess the contribution of donor and/or recipient tissue by performing various combinations and to ...
    X-gal Staining on Adult Mouse Brain Sections
    Authors:  Hiroshi Kokubu and Janghoo Lim, date: 03/05/2014, view: 12858, Q&A: 0
    [Abstract] Knowing expression patterns of given proteins is very important to understand their functions. Immunostaining analysis with specific antibodies is commonly used to identify cells or tissues expressing proteins of interest. Although this technique is regularly used, it requires high quality of specific antibodies and there is no good quality of ...
    Optical Clearing Using SeeDB
    Authors:  Meng-Tsen Ke, Satoshi Fujimoto and Takeshi Imai, date: 02/05/2014, view: 19544, Q&A: 1
    [Abstract] We describe a water-based optical clearing agent, SeeDB (See Deep Brain), which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. SeeDB is a saturated solution of fructose (80.2% w/w) in water with 0.5% α-thioglycerol. In standard SeeDB optical ...
    Dissection of Different Areas from Mouse Hippocampus
    Author:  Faraz A. Sultan, date: 11/05/2013, view: 16523, Q&A: 0
    [Abstract] The hippocampus modulates a number of modules including memory consolidation, spatial navigation, temporal processing and emotion. A banana-shaped structure, the hippocampus is constituted of morphologically distinct subregions including the dentate gyrus, CA3 and CA1 (here, we do not distinguish the “hippocampus proper” which consists only of ...

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