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Detecting Bacterial Surface Organelles on Single Cells using Optical Tweezers
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
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2016 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 32, no 18, 4521-4529 p.Article in journal (Refereed) Published
Abstract [en]

Bacterial cells display a diverse array of surface organelles that are important for a range of processes such as: intercellular communication, motility and adhesion leading to biofilm formation, infections and bacterial spread. More specifically, attachment to host cells by Gram-negative bacteria are mediated by adhesion pili, which are nm wide and µm long fibrous organelles. Since these pili are significantly thinner than the wavelength of visible light, they cannot be detected using standard light microscopy techniques. At present, there is no fast and simple method available to investigate if a single cell expresses pili while keeping the cell alive for further studies. In this study, we present a method to determine the presence of pili on a single bacterium. The protocol involves imaging the bacterium to measure its size, followed by predicting the fluid drag based on its size using an analytical model, and thereafter oscillating the sample while a single bacterium is trapped by an optical tweezer to measure its effective fluid drag. Comparison between the predicted and the measured fluid drag thereby indicate the presence of pili. Herein, we verify the method using polymer coated silica microspheres and Escherichia coli bacteria expressing adhesion pili. Our protocol, can in real time and within seconds assist single cell studies by distinguishing between piliated and non-piliated bacteria.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2016. Vol. 32, no 18, 4521-4529 p.
National Category
Physical Chemistry Materials Engineering
Identifiers
URN: urn:nbn:se:umu:diva-119441DOI: 10.1021/acs.langmuir.5b03845ISI: 000375809100015PubMedID: 27088225OAI: oai:DiVA.org:umu-119441DiVA: diva2:920853
Funder
Swedish Research Council, 2013-5379
Available from: 2016-04-19 Created: 2016-04-19 Last updated: 2016-06-23Bibliographically approved

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Zakrisson, JohanSingh, BhupenderSvenmarker, PontusWiklund, KristerZhang, HanqingHakobyan, ShoghikRamstedt, MadeleineAndersson, Magnus
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