TY - JOUR
T1 - A sensitive, versatile microfluidic assay for bacterial chemotaxis
AU - Mao, Hanbin
AU - Cremer, Paul S.
AU - Manson, Michael D.
PY - 2003/4/29
Y1 - 2003/4/29
N2 - We have developed a microfluidic assay for bacterial chemotaxis in which a gradient of chemoeffectors is established inside a micro-channel via diffusion between parallel streams of liquid in laminar flow. The random motility and chemotactic responses to L-aspartate, L-serine, L-leucine, and Ni2+ of WT and chemotactic-mutant strains of Escherichia coli were measured. Migration of the cells was quantified by counting the cells accumulating in each of 22 outlet ports. The sensitivity of the assay is attested to by the significant response of WT cells to 3.2 nM L-aspartate, a concentration three orders of magnitude lower than the detection limit in the standard capillary assay. The response to repellents was as robust and easily recorded as the attractant response. A surprising discovery was that L-leucine is sensed by Tar as an attractant at low concentrations and by Tsr as a repellent at higher concentrations. This assay offers superior performance and convenience relative to the existing assays to measure bacterial tactic responses, and it is flexible enough to be used in a wide range of different applications.
AB - We have developed a microfluidic assay for bacterial chemotaxis in which a gradient of chemoeffectors is established inside a micro-channel via diffusion between parallel streams of liquid in laminar flow. The random motility and chemotactic responses to L-aspartate, L-serine, L-leucine, and Ni2+ of WT and chemotactic-mutant strains of Escherichia coli were measured. Migration of the cells was quantified by counting the cells accumulating in each of 22 outlet ports. The sensitivity of the assay is attested to by the significant response of WT cells to 3.2 nM L-aspartate, a concentration three orders of magnitude lower than the detection limit in the standard capillary assay. The response to repellents was as robust and easily recorded as the attractant response. A surprising discovery was that L-leucine is sensed by Tar as an attractant at low concentrations and by Tsr as a repellent at higher concentrations. This assay offers superior performance and convenience relative to the existing assays to measure bacterial tactic responses, and it is flexible enough to be used in a wide range of different applications.
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U2 - 10.1073/pnas.0931258100
DO - 10.1073/pnas.0931258100
M3 - Article
C2 - 12704234
AN - SCOPUS:0037965621
SN - 0027-8424
VL - 100
SP - 5449
EP - 5454
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 9
ER -