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ATAC-seq on Sorted Adult Mouse Neurons

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Ex vivo Drosophila Wing Imaginal Disc Culture and Furin Inhibitor Assay

Authors: Alex Sohr, Lijuan Du and Sougata Roy, date: 08/20/2019, view: 348, Q&A: 0

[Abstract] Furin is an evolutionarily conserved proprotein convertase (PC) family enzyme with a broad range of substrates that are essential for developmental, homeostatic, and disease pathways. Classical genetic approaches and in vitro biochemical or cell biological assays identified that precursor forms of most growth factor family proteins are ...

A Novel Technique for Imaging and Analysis of Hair Cells in the Organ of Corti Using Modified Sca/eS and Machine Learning

Authors: Shinji Urata, Tadatsune Iida, Yuri Suzuki, Shiou-Yuh Lin, Yu Mizushima, Chisato Fujimoto, Yu Matsumoto and Tatsuya Yamasoba, date: 08/20/2019, view: 479, Q&A: 0

[Abstract] Here, we describe a sorbitol-based optical clearing method, called modified Sca/eS that can be used to image all hair cells (HCs) in the mouse cochlea. This modification of Sca/eS is defined by three steps: decalcification, de-lipidation, and refractive index matching, which can all be completed within 72 h. Furthermore, we established automated ...

Characterization of Mouse Adult Testicular Macrophage Populations by Immunofluorescence Imaging and Flow Cytometry

Authors: Noushin Mossadegh-Keller and Michael H. Sieweke, date: 03/05/2019, view: 1243, Q&A: 0

[Abstract] Testicular macrophages (tMΦ) are the most abundant immune cells residing in the testis, an immune-privileged organ. TMΦ are known to exhibit different functions, such as protecting spermatozoa from auto-immune attack by producing immunosuppressive cytokines and trophic roles in supporting spermatogenesis and male sex hormone production. They also ...

Blinded Visual Scoring of Images Using the Freely-available Software Blinder

Authors: Steven D. Cothren, Joel N. Meyer and Jessica H. Hartman, date: 12/05/2018, view: 1490, Q&A: 0

[Abstract] In nearly all subfields of biomedical sciences, there are phenotypes that are currently classified by expert visual scoring. In research applications, these classifications require the experimenter to be blinded to the treatment group in order to avoid unintentional bias in scoring. Currently, many labs either use laborious and tedious methods to ...

Staining the Germline in Live Caenorhabditis elegans: Overcoming Challenges by Applying a Fluorescent-dye Feeding Strategy

Author: K. Adam Bohnert, date: 11/05/2018, view: 1892, Q&A: 0

[Abstract] C. elegans provides a tractable model organism for studying germline cell biology. Microscopy experiments are relatively facile, as this worm is transparent and germline development can be observed in real-time using DIC microscopy and/or fluorescent transgenes. Despite these many tools, robust staining techniques for imaging germ cells ...

Detection and Differentiation of Multiple Viral RNAs Using Branched DNA FISH Coupled to Confocal Microscopy and Flow Cytometry

Authors: Nicholas van Buuren and Karla Kirkegaard, date: 10/20/2018, view: 1766, Q&A: 0

[Abstract] Due to the exceptionally high mutation rates of RNA-dependent RNA polymerases, infectious RNA viruses generate extensive sequence diversity, leading to some of the lowest barriers to the development of antiviral drug resistance in the microbial world. We have previously discovered that higher barriers to the development of drug resistance can be ...

Live Confocal Imaging of Brachypodium Spikelet Meristems

Author: Devin Lee O'Connor, date: 09/20/2018, view: 1959, Q&A: 0

[Abstract] Live confocal imaging of fluorescent reporters and stains in plant meristems provides valuable measurements of gene expression, protein dynamics, cell polarity, cell division, and growth. The spikelet meristem in the grass Brachypodium distachyon (Brachypodium) is well suited to live imaging because of the ease of dissection, small ...

Detection of Apoptosis-like Cell Death in Ustilago maydis by Annexin V-FITC Staining

Authors: Dibya Mukherjee, Aroni Mitra and Anupama Ghosh, date: 08/05/2018, view: 1974, Q&A: 0

[Abstract] Programmed cell death (PCD) guides the transition between key developmental stages in many organisms. PCD also remains an important fate for many organisms upon exposure to different stress conditions. Therefore, an insight into the progression of PCD during the execution of a biological phenomenon can yield significant details of the underlying ...

Structural Analysis of Bordetella pertussis Biofilms by Confocal Laser Scanning Microscopy

Authors: Natalia Cattelan, Osvaldo Miguel Yantorno and Rajendar Deora, date: 08/05/2018, view: 2053, Q&A: 0

[Abstract] Biofilms are sessile communities of microbial cells embedded in a self-produced or host-derived exopolymeric matrix. Biofilms can both be beneficial or detrimental depending on the surface. Compared to their planktonic counterparts, biofilm cells display enhanced resistance to killing by environmental threats, chemicals, antimicrobials and host ...

FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans

Authors: Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 3173, Q&A: 0

[Abstract] FRAP (Fluorescence Recovery After Photobleaching) is probably the most direct method to investigate the dynamics of molecules in living cells. Here, we describe FRAP to quantify membrane fluidity in C. elegans. Using FRAP, we have shown that cold, glucose and exogenous saturated fatty acids can decrease the fluidity of cellular membranes ...

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