Investigation of subcellular localization and dynamics of membrane proteins in living bacteria by combining optical micromanipulation and high-resolution microscopy
(Conference Presentation)

Álvaro Barroso Peña; Marcos Nieves; Konrad Teper; Roland Wedlich-Soldner; Cornelia Denz

doi:10.1117/12.2239082

10 November 2016 Investigation of subcellular localization and dynamics of membrane proteins in living bacteria by combining optical micromanipulation and high-resolution microscopy (Conference Presentation)

Álvaro Barroso Peña, Marcos Nieves, Konrad Teper, Roland Wedlich-Soldner, Cornelia Denz

Author Affiliations +

Proceedings Volume 9922, Optical Trapping and Optical Micromanipulation XIII; 99220Y (2016) https://doi.org/10.1117/12.2239082
Event: SPIE Nanoscience + Engineering, 2016, San Diego, California, United States

Abstract

The plasma membrane serves as protective interface between cells and their environment. It also constitutes a hub for selective nutrient uptake and signal transduction. Increasing evidence over the last years indicates that, similar to eukaryotic cells, lateral membrane organization plays an important role in the regulation of prokaryotic signaling pathways. However, the mechanisms underlying this phenomenon are still poorly understood. Spatiotemporal characterization of bacterial signal transduction demands very sensitive high-resolution microscopy techniques due to the low expression levels of most signaling proteins and the small size of bacterial cells. In addition, direct study of subcellular confinement and dynamics of bacterial signaling proteins during the different stages of the signal transduction also requires immobilization in order to avoid cell displacement caused by Brownian motion, local fluid flows and bacterial self-propulsion. In this work we present a novel approach based on the combination of high resolution imaging and optical manipulation that enables the investigation of the distribution and dynamics of proteins at the bacterial plasma membrane. For this purpose, we combine the versatility of holographic optical tweezers (HOT) with the sensitivity and resolution of total internal reflection fluorescence (TIRF) microscopy. Furthermore, we discuss the implementation of microfluidic devices in our integrated HOT+TIRF system for the control of growth conditions of bacterial cells. The capabilities of our workstation provides thus new valuable insights into the fundamental cellular and physical mechanisms underlying the regulation of bacterial signal transduction.

Conference Presentation

Citation Download Citation

Álvaro Barroso Peña, Marcos Nieves, Konrad Teper, Roland Wedlich-Soldner, and Cornelia Denz "Investigation of subcellular localization and dynamics of membrane proteins in living bacteria by combining optical micromanipulation and high-resolution microscopy (Conference Presentation)", Proc. SPIE 9922, Optical Trapping and Optical Micromanipulation XIII, 99220Y (10 November 2016); https://doi.org/10.1117/12.2239082

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KEYWORDS

Proteins

Microscopy

Optical micromanipulation

Bacteria

Control systems

Plasma

Optical tweezers

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