Chemical Imaging of Bacterial Biofilm Dynamics

Open-channel microfluidic platform, SR-FTIR spectromicroscopy work better together.

• Berkeley Lab, Lawrence Livermore National Lab, and UC Berkeley
researchers have developed a robust, label-free method to probe the
chemical underpinnings of developing bacterial biofilms—dynamic
communities of cells that stick to other bacteria or surfaces in water.
• Coupling synchrotron radiation–based Fourier transform infrared (SR-
FTIR) spectromicroscopy from ALS Beamline 1.4.3 with the first open–
channel microfluidic platform could impact several scientific disciplines.
• SR-FTIR spectromicroscopy continuously monitors the changing contents
in living samples without labeling, a technique used here to study dynamic
processes in bacteria living in aqueous environments as they respond to
stimuli and form evolving biofilms.
• The open-channel microfluidic platform maintains the functionality of living
cells while enabling high-quality SR-FTIR spectromicroscopy. It minimizes
IR signal interference by controlling the water film dimensions. A cartoon of the open-channel microfluidic
platform used with SR-FTIR spectromicroscopy to
• To evaluate this technique’s potential, researchers studied: study living bacteria (insert) in aqueous
environments. A modulated IR microbeam is
• Antibiotic resistance in biofilms. By maintaining living bacteria in focused to several micrometers in diameter onto a
biofilm over a long period of time, researchers captured molecular microfluidic platform. The platform consists of a 1-
and structural changes in E. coli biofilms during adaptation to cm-diameter silicon chip (light green) that has
mitomycin antibiotics, a potent DNA crosslinker. been subjected to deep reactive ion etching to
form a hydrophilic microchannel or microwell. The
• Influence of nutrient supply and waste removal on biofilm formation reservoir to the right of the silicon chip provides
in microscopic space. By comparing spectra recorded repeatedly media. The flow rate can be controlled by the
hydrostatic pressure at the inlet and the capillary
for hours, researchers found glycocalyx carbohydrate, a marker of pull at the outlet. The microfluidic platform
biofilm formation, to be limited at sites of low mass exchange while provides enough water to sustain the living cells
exhibiting a cyclic biofilm formation/detachment behavior. without interference on mid-IR spectroscopy.

H-Y. Holman, et al., Anal. Chem. 81, (20) 8564 (2010) •

http://www-als.lbl.gov/index.php/science-highlights/science-highlights/466