Cell Biology

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    Protocols in Current Issue
    A Flow Cytometric Method to Determine Transfection Efficiency
    [Abstract] Mammalian cell transfection is a powerful technique commonly used in molecular biology to express exogenous DNA or RNA in cells and study gene and protein function. Although several transfection strategies have been developed, there is a wide variation with regards to transfection efficiency, cell toxicity and reproducibility. Thus, a sensitive ...
    High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level
    Authors:  Thomas Luh, Kimberly Lucero, Wenji Ma, Jaeyop Lee, Yu Jerry Zhou, Yufeng Shen and Kang Liu, date: 05/20/2018, view: 2082, Q&A: 0
    [Abstract] The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to evaluate high-dimensional transcriptomic and proteomic data (relating to cellular mRNA and protein), ...
    Determining Genome Size from Spores of Seedless Vascular Plants
    Authors:  Li-Yaung Kuo and Yao-Moan Huang , date: 06/05/2017, view: 4027, Q&A: 0
    [Abstract] Seedless vascular plants, including ferns and lycophytes, produce spores to initiate the gametophyte stage and to complete sexual reproduction. Approximately 10% of them are apomictic through the production of genomic unreduced spores. Being able to measure the spore nuclear DNA content is therefore important to infer their reproduction mode. Here ...
    Flow Cytometric Analysis of HIV-1 Transcriptional Activity in Response to shRNA Knockdown in A2 and A72 J-Lat Cell Lines
    Authors:  Daniela Boehm and Melanie Ott, date: 06/05/2017, view: 4253, Q&A: 0
    [Abstract] The main obstacle to eradicating HIV-1 from patients is post-integration latency (Finzi et al., 1999). Antiretroviral treatments target only actively replicating virus, while latent infections that have low or no transcriptional activity remain untreated (Sedaghat et al., 2007). To eliminate viral reservoirs, one strategy focuses ...
    Flow Cytometric Analysis of Drug-induced HIV-1 Transcriptional Activity in A2 and A72 J-Lat Cell Lines
    Authors:  Daniela Boehm and Melanie Ott, date: 05/20/2017, view: 5482, Q&A: 0
    [Abstract] The main obstacle to eradicating HIV-1 from patients is post-integration latency (Finzi et al., 1999). Antiretroviral treatments target only actively replicating virus, while latent infections that have low or no transcriptional activity remain untreated (Sedaghat et al., 2007). A combination of antiretroviral treatments with ...
    Quantitative Analysis of Exosome Secretion Rates of Single Cells
    Authors:  Yu-Jui Chiu, Wei Cai, Tiffany Lee, Julia Kraimer and Yu-Hwa Lo, date: 02/20/2017, view: 6310, Q&A: 0
    [Abstract] To study the inhomogeneity within a cell population including exosomes properties such as exosome secretion rate of cells and surface markers carried by exosomes, we need to quantify and characterize those exosomes secreted by each individual cell. Here we develop a method to collect and analyze exosomes secreted by an array of single cells using ...
    Protocol for Microfluidic System to Automate the Preparation and Fractionation of the Nucleic Acids in the Cytoplasm Versus Nuclei of Single Cells
    Authors:  Kentaro Kuriyama, Hirofumi Shintaku and Juan G. Santiago, date: 06/20/2016, view: 4275, Q&A: 0
    [Abstract] This protocol describes the extraction, fractionation, and recovery of cytoplasmic nucleic acids (e.g., cytoplasmic RNA) versus nucleic acids in the cell nucleus (including genomic DNA, gDNA) from single cells with a microfluidic system. The method enables independent, sequence-specific analyses of these critical markers (Kuriyama et al. ...
    Phenotyping of Live Human PBMC using CyTOFTM Mass Cytometry
    Authors:  Michael D. Leipold and Holden Maecker, date: 01/20/2015, view: 9774, Q&A: 3
    [Abstract] Single-cell analysis has become an method of importance in immunology. Fluorescence flow cytometry has been a major player. However, due to issues such as autofluorescence and emission spillover between different fluorophores, alternative techniques are being developed. In recent years, mass cytometry has emerged, wherein antibodies labeled with ...
    Adenosine A2A Receptor Ligand Binding Experiments by Using Real-time Single-cell FRET
    Authors:  Víctor Fernández-Dueñas, Kenneth A. Jacobson and Francisco Ciruela, date: 03/20/2014, view: 6830, Q&A: 0
    [Abstract] We designed a fluorescence resonance energy transfer (FRET)-based approach to study the ligand binding constants of the adenosine A2A receptor (A2AR). Our assay is based in the interaction of a fluorescent A2AR agonist ligand (MRS5424) with an A2AR tagged with the cyan fluorescent protein (CFP) at the ...
    Neutral Comet Assay
    Authors:  Elisa Boutet-Robinet, Didier Trouche and Yvan Canitrot, date: 09/20/2013, view: 20060, Q&A: 1
    [Abstract] The Comet assay (or Single Cell Gel Electrophoresis assay) is a sensitive technique to detect DNA damage at the level of an individual cell. This technique is based on micro-electrophoresis of cells DNA content. Briefly, cells are embedded in agarose, lysed and submitted to an electric field, before the staining step with a fluorescent DNA binding ...



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