Mammalia

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    Protocols in Current Issue
    Bone-in-culture Array to Model Bone Metastasis in ex vivo Condition
    Authors:  Hai Wang and Xiang H.-F. Zhang, date: 01/20/2020, view: 96, Q&A: 0
    Bone is the most frequently affected organ by metastases of breast cancer and prostate cancer. Our knowledge on bone metastasis is extremely limited due to the lack of potent and efficient experimental models. We developed the “Bone-In-Culture Array ...
    Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
    Authors:  Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 71, Q&A: 0
    Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated ...
    Protocols for Processing and Interpreting cryoEM Data Using Bsoft: A Case Study of the Retinal Adhesion Protein, Retinoschisin
    Author:  J. Bernard Heymann, date: 01/20/2020, view: 105, Q&A: 0
    The goal of cryoEM is to determine the structures of biomolecules from electron micrographs. In many cases the processing is straightforward and can be handled with routine protocols. In other cases, the properties and behavior of the specimen ...
    Isolation of Stem Cells, Endothelial Cells and Pericytes from Human Infantile Hemangioma
    Authors:  Lan Huang and Joyce Bischoff, date: 01/20/2020, view: 127, Q&A: 0
    Infantile hemangioma (IH) is a vascular tumor noted for its excessive blood vessel formation during infancy, glucose-transporter-1 (GLUT1)-positive staining of the blood vessels, and its slow spontaneous involution over several years in early ...
    Quantifying HIV-1-Mediated Gut CD4+ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model
    Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture ...
    A Simple and Efficient Method for Concomitant Isolation and Culture of Enriched Astroglial and Microglial Cells from the Rat Spinal Cord
    Authors:  Pooja Shree Mishra and Trichur R Raju, date: 01/20/2020, view: 188, Q&A: 0
    Investigations into glial biology have contributed substantially in understanding the physiology and pathology of the nervous system. However, intricacies of the neuron-glial and glial-glial interactions in vivo present significant ...
    Protocol for Measuring Free (Low-stress) Exploration in Rats
    Authors:  Wojciech Pisula and Klaudia Modlinska, date: 01/20/2020, view: 152, Q&A: 0
    Research on exploratory behavior plays a key part in behavioral science. Studying exploratory behavior of laboratory rodents may provide important data about many developmental and neurobiological processes occurring in animal ontogenesis. The ...
    Study of Microbial Extracellular Vesicles: Separation by Density Gradients, Protection Assays and Labelling for Live Tracking
    Extracellular vesicles (EVs) are produced by all domains of life including Bacteria, Archaea and Eukarya. EVs are critical for cellular physiology and contain varied cargo: virulence factors, cell wall remodeling enzymes, extracellular matrix ...
    Studying Protein Aggregation in the Context of Liquid-liquid Phase Separation Using Fluorescence and Atomic Force Microscopy, Fluorescence and Turbidity Assays, and FRAP
    Authors:  W. Michael Babinchak and Witold K. Surewicz, date: 01/20/2020, view: 165, Q&A: 0
    Liquid-liquid phase separation (LLPS) underlies the physiological assembly of many membrane-less organelles throughout the cell. However, dysregulation of LLPS may mediate the formation of pathological aggregates associated with neurodegenerative ...
    Characterizing the Two-photon Absorption Properties of Fluorescent Molecules in the 680-1300 nm Spectral Range
    Authors:  Mikhail Drobizhev, Rosana S Molina and Thomas E Hughes, date: 01/20/2020, view: 71, Q&A: 0
    Two-photon laser scanning microscopy (2PLSM) is a state-of-the-art technique used for non-invasive imaging deep inside the tissue, with high 3D resolution, minimal out-of-focus photodamage, and minimal autofluorescence background. For optimal ...
    Ribonucleoprotein Immunoprecipitation (RIP) Analysis
    Authors:  Jennifer L. Martindale, Myriam Gorospe and Maria L. Idda, date: 01/20/2020, view: 197, Q&A: 0
    RNAs and RNA-binding proteins (RBPs) can interact dynamically in ribonucleoprotein (RNP) complexes that play important roles in controlling gene expression programs. One of the powerful ways to investigate changes in the association of RNAs with an ...
    A Single Test to Study Social Behavior and Repetitive Self-grooming in Mice
    Authors:  Niraj V. Lawande, Ammar L. Ujjainwala and Catherine A. Christian, date: 01/20/2020, view: 128, Q&A: 0
    The ability to recognize and interact with members of the same species is essential for social communication. Investigating the neural substrates of social interest and recognition may offer insights into the behavioral differences present in ...
    Bone-in-culture Array to Model Bone Metastasis in ex vivo Condition
    Authors:  Hai Wang and Xiang H.-F. Zhang, date: 01/20/2020, view: 96, Q&A: 0
    [Abstract] Bone is the most frequently affected organ by metastases of breast cancer and prostate cancer. Our knowledge on bone metastasis is extremely limited due to the lack of potent and efficient experimental models. We developed the “Bone-In-Culture Array (BICA)” platform to model the bone colonization of cancer cells in ex vivo cultures. The ...
    Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
    Authors:  Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 71, Q&A: 0
    [Abstract] Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated by pathogens. Dorsal ruffles and their resulting macropinosomes are key sites for environmental ...
    Protocols for Processing and Interpreting cryoEM Data Using Bsoft: A Case Study of the Retinal Adhesion Protein, Retinoschisin
    Author:  J. Bernard Heymann, date: 01/20/2020, view: 105, Q&A: 0
    [Abstract] The goal of cryoEM is to determine the structures of biomolecules from electron micrographs. In many cases the processing is straightforward and can be handled with routine protocols. In other cases, the properties and behavior of the specimen require adaptions to properly interpret the data. Here I describe the protocols for examining the higher ...
    Isolation of Stem Cells, Endothelial Cells and Pericytes from Human Infantile Hemangioma
    Authors:  Lan Huang and Joyce Bischoff, date: 01/20/2020, view: 127, Q&A: 0
    [Abstract] Infantile hemangioma (IH) is a vascular tumor noted for its excessive blood vessel formation during infancy, glucose-transporter-1 (GLUT1)-positive staining of the blood vessels, and its slow spontaneous involution over several years in early childhood. For most children, IH poses no serious threat because it will eventually involute, but a subset ...
    Quantifying HIV-1-Mediated Gut CD4+ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model
    [Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...
    A Simple and Efficient Method for Concomitant Isolation and Culture of Enriched Astroglial and Microglial Cells from the Rat Spinal Cord
    Authors:  Pooja Shree Mishra and Trichur R Raju, date: 01/20/2020, view: 188, Q&A: 0
    [Abstract] Investigations into glial biology have contributed substantially in understanding the physiology and pathology of the nervous system. However, intricacies of the neuron-glial and glial-glial interactions in vivo present significant challenges while delineating the individual cell-type contributions, thus making the in vitro ...
    Protocol for Measuring Free (Low-stress) Exploration in Rats
    Authors:  Wojciech Pisula and Klaudia Modlinska, date: 01/20/2020, view: 152, Q&A: 0
    [Abstract] Research on exploratory behavior plays a key part in behavioral science. Studying exploratory behavior of laboratory rodents may provide important data about many developmental and neurobiological processes occurring in animal ontogenesis. The proposed protocol for measuring the free (low-stress) exploration behavior in rats is straightforward, ...
    Study of Microbial Extracellular Vesicles: Separation by Density Gradients, Protection Assays and Labelling for Live Tracking
    [Abstract] Extracellular vesicles (EVs) are produced by all domains of life including Bacteria, Archaea and Eukarya. EVs are critical for cellular physiology and contain varied cargo: virulence factors, cell wall remodeling enzymes, extracellular matrix components and even nucleic acids and metabolites. While various protocols for isolating EVs have been ...
    Studying Protein Aggregation in the Context of Liquid-liquid Phase Separation Using Fluorescence and Atomic Force Microscopy, Fluorescence and Turbidity Assays, and FRAP
    Authors:  W. Michael Babinchak and Witold K. Surewicz, date: 01/20/2020, view: 165, Q&A: 0
    [Abstract] Liquid-liquid phase separation (LLPS) underlies the physiological assembly of many membrane-less organelles throughout the cell. However, dysregulation of LLPS may mediate the formation of pathological aggregates associated with neurodegenerative diseases. Here, we present complementary experimental approaches to study protein aggregation within ...
    Characterizing the Two-photon Absorption Properties of Fluorescent Molecules in the 680-1300 nm Spectral Range
    Authors:  Mikhail Drobizhev, Rosana S Molina and Thomas E Hughes, date: 01/20/2020, view: 71, Q&A: 0
    [Abstract] Two-photon laser scanning microscopy (2PLSM) is a state-of-the-art technique used for non-invasive imaging deep inside the tissue, with high 3D resolution, minimal out-of-focus photodamage, and minimal autofluorescence background. For optimal application of fluorescent probes in 2PLSM, their two-photon absorption (2PA) spectra, expressed in ...



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