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JoVE Journal

Immunology and Infection

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Applying Live Cell Imaging and Cryo-Electron Tomography to Resolve Spatiotemporal Features of the Legionella pneumophila Dot/Icm Secretion System

6.7K Views

09:12 min

March 10th, 2020

March 10th, 2020

6,710 Views

0:04

Introduction

0:55

Live Cell Imaging of L. pneumophila with Fluorescently Tagged Dot/Icm Components

2:47

Quantification of Polar Localization and Dynamics of Dot/Icm Components

5:36

Detection of sfGFP Mass Density with Cryo-ET

6:23

Results: Imaging Dot/Icm Secretion Systems in Intact Bacteria

8:17

Conclusion

副本

This protocol is useful for characterizing the assembling functions of bacterial secretion complexes and can lead to a fundamental understanding of the molecular mechanisms that are required for host pathogen interactions. This technique integrate

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Imaging of bacterial cells is an emerging systems biology approach focused on defining static and dynamic processes that dictate the function of large macromolecular machines. Here, integration of quantitative live cell imaging and cryo-electron tomography is used to study Legionella pneumophila type IV secretion system architecture and functions.

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