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European Microscopy Congress 2020 Abstract Database

Quantification of E. coli invasion into eukaryotic cells by Imaging flow cytometry.

Quantification of E. coli invasion into eukaryotic cells by Imaging flow cytometry.

Session

LSA.3 - Applications for imaging sub-cellular events at high resolution

Authors

Mr Rasaq Akinsola (1), Associate Professor Kumaran Narayanan (1)

Affiliations

1. Monash University Malaysia

Keywords

 Invasion, quantification, E. coli, adhesion, Imaging flow cytometry, Simultaneous

Abstract text

Bacteria gene transfer is a useful technique that offers an attractive alternative to virus-based methods for applications in DNA vaccination, cancer and gene therapy. However, the invasiveness of the vector needs to be precisely and accurately measured to study its intracellular trafficking. Traditionally, the antibiotic protection assay is used to estimate the internalized bacteria by agar plating and colony counting, but this method lacks reliability and reproducibility. Here, we described a rapid method to determine bacteria vector adhesion and invasion into mammalian cells with an E. coli vector that expresses red fluorescence protein using Imaging flow cytometry. This method was performed on MCF-7, A549, and HEK-293 cells. We obtained high cellular infection of up to 70.47 % in MCF as compared to A549 and HEK-293 cells, respectively. The Imaging flow cytometry method allows real-time and simultaneous quantification of both bacteria attachment and entry into host cells. This strategy can be applied to other bacteria vectors. 

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