Type

Journal Article

Authors

S G J Smith
J C Simpson
G Galea
S Paré
C M Murphy

Subjects

Biochemistry

Topics
mammalian cells bacteria green fluorescent protein association image bacterial automated analysis expression

Accurate and semi-automated analysis of bacterial association with mammalian cells. (2015)

Abstract To efficiently and accurately quantify the interactions of bacteria with mammalian cells, a reliable fluorescence microscopy assay was developed. Bacteria were engineered to become rapidly and stably fluorescent using Green Fluorescent Protein (GFP) expressed from an inducible Tet promoter. Upon application of the fluorescent bacteria onto a monolayer, extracellular bacteria could be discriminated from intracellular bacteria by antibody staining and microscopy. All bacteria could be detected by GFP expression. External bacteria stained orange, whereas internalised bacteria did not. Internalised bacteria could thus be discriminated from external bacteria by virtue of being green but not orange fluorescent. Image acquisition and counting of various fluorophore-stained entities were accomplished with a high-content screening platform. This allowed for semi-automated and accurate counting of intracellular and extracellular bacteria.
Collections Ireland -> Trinity College Dublin -> PubMed

Full list of authors on original publication

S G J Smith, J C Simpson, G Galea, S Paré, C M Murphy

Experts in our system

1
Stephen Smith
Trinity College Dublin
Total Publications: 27
 
2
Jeremy C. Simpson
University College Dublin
Total Publications: 38