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Superresolution Microscopy of Drosophila Indirect Flight Muscle Sarcomeres

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Isolation and High Throughput Flow Cytometric Apoptosis Assay of Human Neutrophils to Enable Compound Library Screening

Authors: K. D. Herman, A. Rahman and L. R. Prince, date: 06/05/2020, view: 534, Q&A: 0

[Abstract] The study of human neutrophils in vitro is challenging due to their short half-life and propensity for activation. However, with careful handling and manipulation in the laboratory, they can be a powerful tool to investigate immune responses in health and disease. Here we describe a method for the isolation of human neutrophils from ...

Methodology for in vitro Assessment of Human T Cell Activation and Blockade

Authors: Joseph R. Podojil, Igal Ifergan, Ming-Yi Chiang, Joshua J. Meeks and Stephen D. Miller, date: 06/05/2020, view: 595, Q&A: 0

[Abstract] Methods to test both the functionality and mechanism of action for human recombinant proteins and antibodies in vitro have been limited by multiple factors. To test the functionality of a recombinant protein or antibody, the receptor, the receptor-associated ligand, or both must be expressed by the cells present within the in vitro ...

A Modified Semisolid Clonal Culture for Identification of B-1 and B-2 Progenitor Colony Forming Ability of Mouse Embryonic Hemogenic Endothelial Cells

Authors: Michihiro Kobayashi and Momoko Yoshimoto, date: 05/05/2020, view: 659, Q&A: 0

[Abstract] The search for the origin of the first hematopoietic stem cells (HSCs) in the mouse embryo has been a hot topic in the field of developmental hematopoiesis. Detecting lymphoid potential is one of the supportive evidence to show the definitive hematopoietic activity of HSCs. However, the first B-lymphoid potential in the mouse embryos are reported ...

Quantification of Bacteria Residing in Caenorhabditis elegans Intestine

Authors: M. Fernanda Palominos and Andrea Calixto, date: 05/05/2020, view: 833, Q&A: 0

[Abstract] Quantification of intestinal colonization by pathogenic or commensal bacteria constitute a critical part of the analysis to understand host-microbe interactions during different time points of their interplay. Here we detail a method to isolate non-pathogenic and pathogenic bacteria from C. elegans intestines, and classify gut phenotypes ...

Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle

Authors: Diantha van de Vlekkert, Xiaohui Qiu, Ida Annunziata and Alessandra d'Azzo, date: 04/05/2020, view: 577, Q&A: 0

[Abstract] Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have ...

Fractionation of Ovarian Follicles and in vitro Oocyte Maturation and Ovulation Assay of Ciona intestinalis Type A

Authors: Shin Matsubara, Akira Shiraishi, Tomohiro Osugi, Tsuyoshi Kawada and Honoo Satake, date: 04/05/2020, view: 672, Q&A: 0

[Abstract] Ascidians are the closest living relatives of vertebrates (Delsuc et al., 2006; Satoh et al., 2014) and are important for the evolutionary study of the ovarian follicle development including oocyte maturation and ovulation. However, neither the endogenous factors nor the molecular mechanisms underlying the oocyte maturation and ...

Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands

Authors: Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 947, Q&A: 0

[Abstract] Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells suspension with high quality, especially from the primary solid organs like mammary glands. Digestion of ...

RNA Sequencing of Single Myofibers from Mus musculus

Authors: Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 1624, Q&A: 0

[Abstract] Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...

Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus

Authors: Valeria Lallai, Amina Ahmed and Christie D. Fowler, date: 02/20/2020, view: 1001, Q&A: 0

[Abstract] The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular environment. This biofluid contains an enriched source of proteins, ions, and other signaling molecules for ...

Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation

Authors: Gileade P. Freitas, Alann T. P. Souza, Helena B. Lopes, Rayana L. B. Trevisan, Fabiola S. Oliveira, Roger R. Fernandes, Fernanda U. Ferreira, Felipe A. Ros, Marcio M. Beloti and Adalberto L. Rosa, date: 02/20/2020, view: 1039, Q&A: 0

[Abstract] Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and ...

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