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MRC-Laboratory of Molecular Biology, Cambridge, UK

3 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun
Jason Arsenault 1,2, Andras Nagy 1, Jeffrey T. Henderson 1, John A. O'Brien 2
1Leslie Dan Faculty of Pharmacy, University of Toronto, 2MRC-Laboratory of Molecular Biology, Cambridge, UK

Recent improvements in organotypic brain slice preparations have permitted their exploitation for biotechnological applications. Organotypic slices maintain local structural characteristics of in vivo biology, including functional synaptic connections. Here we present a regioselective biolistic delivery method to label and genetically manipulate these slices.

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Biology

High-throughput Measurement of Dictyostelium discoideum Macropinocytosis by Flow Cytometry
Thomas Williams 1, Robert R. Kay 1
1MRC-Laboratory of Molecular Biology, Cambridge, UK

Macropinocytosis, large-scale non-specific fluid uptake, is important in many areas of clinical biology including immunology, infection, cancer, and neurodegenerative diseases. Here, existing techniques have been adapted to allow high-throughput, single-cell resolution measurement of macropinocytosis in the macropinocytosis model organism Dictyostelium discoideum using flow cytometry.

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Genetics

Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria
Peggy Paschke 1, David A. Knecht 2, Thomas D. Williams 1, Peter A. Thomason 3, Robert H. Insall 3, Jonathan R. Chubb 4,5, Robert R. Kay 1, Douwe M. Veltman 1
1MRC Laboratory of Molecular Biology, 2Department of Molecular and Cell Biology, University of Connecticut, 3Cancer Research UK Beatson Institute Glasgow, 4MRC Laboratory for Molecular Cell Biology, University College London, 5Department of Cell and Developmental Biology, University College London

Dictyostelium discoideum is a popular model organism to study complex cellular processes such as cell migration, endocytosis, and development. The utility of the organism is dependent on the feasibility of genetic manipulation. Here, we present methods to transfect Dictyostelium discoideum cells that overcome existing limitations of culturing cells in liquid media.

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