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Latest Articles

Lasers for Flow Cytometry: Current and Future Trends
Howard M. Shapiro, William G. Telford
Lasers are the principal light sources for flow cytometers. Virtually all cytometers are equipped with at least one (and often many more) lasers. This unit covers the various types of lasers available and the qualities that make them suitable or unsuitable for use in flow cytometers. Also included
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Non‐Parametric Comparison of Single Parameter Histograms
James C.S. Wood
A number of methods have been developed to compare single parameter histograms. Some perform a channel‐by‐channel analysis and others give a single statistic about how the histograms may or may not differ. If they do differ, then the significance of the difference or confidence limit is usually
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Generating Quantitative Cell Identity Labels with Marker Enrichment Modeling (MEM)
Kirsten E. Diggins, Jocelyn S. Gandelman, Caroline E. Roe, Jonathan M. Irish
Multiplexed single‐cell experimental techniques like mass cytometry measure 40 or more features and enable deep characterization of well‐known and novel cell populations. However, traditional data analysis techniques rely extensively on human experts or prior knowledge, and novel machine learning
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Basics of Digital Microscopy
Callen T. Wallace, Morgan Jessup, Tytus Bernas, Karina A. Peña, Michael J. Calderon, Patricia A. Loughran
Modern digital microscopy combines the equipment of classical light microscopy with a computerized imaging system. The technique comprises image formation by optics, image registration by a camera, and saving of image data in a computer file. This chapter describes limitations that are particular to
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Live‐Animal Imaging of Renal Function by Multiphoton Microscopy
Kenneth W. Dunn, Timothy A. Sutton, Ruben M. Sandoval
Intravital microscopy, microscopy of living animals, is a powerful research technique that combines the resolution and sensitivity found in microscopic studies of cultured cells with the relevance and systemic influences of cells in the context of the intact animal. The power of intravital
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Transfection by Electroporation
Huntington Potter, Richard Heller
Electroporation—the use of high‐voltage electric shocks to introduce DNA into cells—can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit
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Making Use of Cancer Genomic Databases
Chad J. Creighton
The vast amounts of genomic data now deposited in public repositories represent rich resources for cancer researchers. Large‐scale genomics initiatives such as The Cancer Genome Atlas have made available data from multiple molecular profiling platforms (e.g., somatic mutation, RNA and protein
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Avian Retrovirus‐Mediated Tumor‐Specific Gene Knockout
Wei Wang, Bingning Dong, Feng Yang
The RCAS (replication‐competent avian sarcoma leukosis virus long‐terminal repeat with splice acceptor)‐TVA (tumor virus A) gene delivery system has been successfully used in modeling human cancers. Based on this, we have recently developed a novel RCI‐Oncogene (RCAS‐Cre‐IRES‐Oncogene) gene delivery
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CRISPR/Cas9‐Mediated Genome Editing in Epstein‐Barr Virus‐Transformed Lymphoblastoid B‐Cell Lines
Sizun Jiang, Liang Wei Wang, Michael J. Walsh, Stephen J. Trudeau, Catherine Gerdt, Bo Zhao, Benjamin E. Gewurz
Epstein‐Barr virus (EBV) efficiently transforms primary human B cells into immortalized lymphoblastoid cell lines (LCLs), which are extensively used in human genetic, immunological and virological studies. LCLs provide unlimited sources of DNA for genetic investigation, but can be difficult to
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Modulating Gene Expression in Epstein‐Barr Virus (EBV)‐Positive B Cell Lines with CRISPRa and CRISPRi
Liang Wei Wang, Stephen J. Trudeau, Chong Wang, Catherine Gerdt, Sizun Jiang, Bo Zhao, Benjamin E. Gewurz
Epstein‐Barr virus (EBV) transforms small resting primary B cells into large lymphoblastoid cells which are able to grow and survive in vitro indefinitely. These cells represent a model for oncogenesis. In this unit, variants of conventional clustered regularly interspaced short palindromic repeats
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CRISPR‐Cas9‐Edited Site Sequencing (CRES‐Seq): An Efficient and High‐Throughput Method for the Selection of CRISPR‐Cas9‐Edited Clones
Yaligara Veeranagouda, Delphine Debono‐Lagneaux, Hamida Fournet, Gilbert Thill, Michel Didier
The emergence of clustered regularly interspaced short palindromic repeats–Cas9 (CRISPR‐Cas9) gene editing systems has enabled the creation of specific mutants at low cost, in a short time and with high efficiency, in eukaryotic cells. Since a CRISPR‐Cas9 system typically creates an array of
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Pooled Lentiviral‐Delivery Genetic Screens
Federica Piccioni, Scott T. Younger, David E. Root
Pooled cell‐based screens of mammalian genetic perturbations enable systematic large‐scale, even genome‐scale, evaluation of gene function. Pooled screens introduce genetic perturbations into a cell population through viral transduction such that each cell integrates into its DNA a single or small
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