%0 Online Multimedia
%A Baker, Joshua
D.
%A Kysela, David T.
%A Zhou, Jinsheng
%A Madren, Seth M.
%A Wilkens, Andrew S.
%A Brun, Yves V.
%A Jacobson, Stephen C.
%D 2016
%T Programmable, Pneumatically Actuated Microfluidic
Device with an Integrated Nanochannel Array To Track Development of
Individual Bacteria
%U https://acs.figshare.com/articles/media/Programmable_Pneumatically_Actuated_Microfluidic_Device_with_an_Integrated_Nanochannel_Array_To_Track_Development_of_Individual_Bacteria/3471029
%R 10.1021/acs.analchem.6b00889.s003
%2 https://acs.figshare.com/ndownloader/files/5465504
%K lineage
%K Integrated Nanochannel Array
%K generation
%K Pneumatically Actuated Microfluidic Device
%K nanochannel array
%K device
%X We
describe a microfluidic device with an integrated nanochannel
array to trap individual bacteria and monitor growth and reproduction
of lineages over multiple generations. Our poly(dimethylsiloxane)
device comprises a pneumatically actuated nanochannel array that includes
1280 channels with widths from 600 to 1000 nm to actively trap diverse
bacteria. Integrated pumps and valves perform on-chip fluid and cell
manipulations that provide dynamic control of cell loading and nutrient
flow, permitting chemostatic growth for extended periods of time (typically
12 to 20 h). Nanochannels confine bacterial growth to a single dimension,
facilitating high-resolution, time-lapse imaging and tracking of individual
cells. We use the device to monitor the growth of single bacterial
cells that undergo symmetric (Bacillus subtilis)
and asymmetric (Caulobacter crescentus) division
and reconstruct their lineages to correlate growth measurements through
time and among related cells. Furthermore, we monitor the motility
state of single B. subtilis cells across multiple
generations by the expression of a fluorescent reporter protein and
observe that the state of the epigenetic switch is correlated over
five generations. Our device allows imaging of cellular lineages with
high spatiotemporal resolution
to facilitate the analysis of biological processes spanning multiple
generations.
%I ACS Publications