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Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer

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Using the Parafilm-assisted Microdissection (PAM) Method to Sample Rodent Nucleus Accumbens

Authors: Marco Pagliusi Jr., Arthur F. Brandão, Gabriel G. Zanetti, Ivan J. M. Bonet, Cesar R. Sartori and Andre S. Vieira, date: 12/05/2020, view: 420, Q&A: 0

[Abstract] Microdissection techniques are very important for anatomical and functional studies focused on neuroscience, where it is often necessary microdissect specific brain areas to perform molecular or anatomical analyses. The parafilm^®-assisted microdissection (PAM) was previously described and involves the microdissection of tissue ...

Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands

Authors: Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 1766, 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 ...

Acute Cerebellar Slice Preparation Using a Tissue Chopper

Authors: Yubin Wang and Michel Baudry, date: 03/05/2019, view: 2903, Q&A: 0

[Abstract] Acute cerebellar slices are widely used among neuroscientists to study the properties of excitatory and inhibitory synaptic transmission as well as intracellular signaling pathways involved in their regulation in cerebellum. The cerebellar cortex presents a well-organized circuitry, and several neuronal pathways can be stimulated and recorded ...

Identification and Quantitation of Leukocyte Populations in Human Kidney Tissue by Multi-parameter Flow Cytometry

Authors: Katrina Kildey, Becker M.P. Law, Kimberly A. Muczynski, Ray Wilkinson, Helen Healy and Andrew J. Kassianos, date: 08/20/2018, view: 5184, Q&A: 0

[Abstract] Inflammatory immune cells play direct pathological roles in cases of acute kidney injury (AKI) and chronic kidney disease (CKD). However, the identification and characterization of distinct populations of leukocytes in human kidney biopsies have been confounded by the limitations of immunohistochemical (IHC)-based techniques used to detect them. ...

In vitro Explant Cultures to Interrogate Signaling Pathways that Regulate Mouse Lung Development

Authors: Changfu Yao, Gianni Carraro and Barry R. Stripp, date: 05/20/2018, view: 4373, Q&A: 0

[Abstract] Early mouse lung development, including specification of primordia, patterning of early endoderm and determination of regional progenitor cell fates, is tightly regulated. The ability to culture explanted embryonic lung tissue provides a tractable model to study cellular interactions and paracrine factors that regulate these processes. We provide ...

Organotypic Explants of the Embryonic Rodent Hippocampus: An Accessible System for Transgenesis

Authors: Archana Iyer , Lakshmi Subramanian and Shubha Tole, date: 03/20/2018, view: 4482, Q&A: 0

[Abstract] This protocol describes the technique of ex-vivo electroporation to target embryonic hippocampal progenitors in an organotypic slice preparation. This technique allows gene perturbation for examining developmental processes in the embryonic hippocampus while retaining the environment and connectivity of the cells. Gene perturbation can ...

Obtaining Acute Brain Slices

Authors: Thomas Papouin and Philip G. Haydon, date: 01/20/2018, view: 14362, Q&A: 1

[Abstract] Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step process are scarce, in particular for routine acute preparations such as from the mouse hippocampus. Here we provide a detailed protocol for the ...

Ex vivo Trophoblast-specific Genetic Manipulation Using Lentiviral Delivery

Authors: Damayanti Chakraborty, Masanaga Muto and Michael J. Soares, date: 12/20/2017, view: 4623, Q&A: 0

[Abstract] In this protocol report, we describe a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Lentiviral packaged gene constructs can be efficiently and specifically delivered to the trophoblast cell lineage of the blastocyst. The consequences of ‘gain-of-function’ and ‘loss-of-function’ blastocyst ...

Mandibular Explant Assay for Investigating Extrinsic Stimuli on Bone and Cartilage Development

Authors: Jamie Jiang, Jessica W. Bertol and Walid D. Fakhouri, date: 12/05/2017, view: 4954, Q&A: 0

[Abstract] A major issue in developmental biology is to determine how embryonic tissues respond to molecular signals in a timely manner and given the position-restricted instructions during morphogenesis, of which Meckel’s cartilage is a classical example. The ex-vivo explant model is a practical and convenient system that allows investigation of ...

Quantification of Trypanosoma cruzi in Tissue and Trypanosoma cruzi Killing Assay

Authors: Hisako Kayama, Shoko Kitada and Kiyoshi Takeda, date: 11/20/2017, view: 5221, Q&A: 0

[Abstract] Infection with Trypanosoma cruzi causes Chagas disease. The methods provided here allow for the quantification of T. cruzi in the liver, heart, and blood of intraperitoneally-infected mice and analysis of the killing activity of the cells infected with T. cruzi in vitro.

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