The
ability to observe dynamic chemical processes (e.g., signaling,
transport, etc.) in vivo or in situ using nondestructive chemical imaging opens a new door to understanding
the complex dynamics of developing biological systems. With the advent
of “biology-on-a-chip” devices has come the ability
to monitor dynamic chemical processes in a controlled environment,
using these engineered habitats to capture key features of natural
systems while allowing visual observation of system development. Having
the capability to spatially and temporally map the chemical signals
within these devices may yield new insights into the forces that drive
biosystem development. Here, a porous membrane sealed microfluidic
device was designed to allow normal microfluidic operation while enabling
continuous, location specific sampling and chemical characterization
by liquid microjunction surface sampling probe mass spectrometry (LMJ-SSP
MS). LMJ-SSP was used to extract fluids with nL-to-μL/min flow
rates directly from selected areas of the microfluidic device without
negatively impacting the device function. These extracts were subsequently
characterized using MS. This technique was used to acquire MS images
of the entirety of several multi-input microfluidic devices having
different degrees of fluid mixing. LMJ-SSP MS imaging visualized the
spatial distribution of chemical components within the microfluidic
channels and could visualize chemical reactions occurring in the device.
These microfluidic devices with a porous membrane wall are wholly
compatible with the construction of biology-on-a-chip devices. This
ultimately would enable correlation of biosystem physical structure
with an evolving chemical environment
A soil-mimicking rhizosphere-on-a-chip is amenable for long-term plant growth and enables simulation of root exudate diffusion and experimental validation of carbon hotspot formation from the interaction between roots and the synthetic soil grains.
The plant rhizosphere is a complex and dynamic chemical
environment
where the exchange of molecular signals between plants, microbes,
and fungi drives the development of the entire biological system.
Exogenous compounds in the rhizosphere are known to affect plant-microbe
organization, interactions between organisms, and ultimately, growth
and survivability. The function of exogenous compounds in the rhizosphere
is still under much investigation, specifically with respect to their
roles in plant growth and development, the assembly of the associated
microbial community, and the spatiotemporal distribution of molecular
components. A major challenge for spatiotemporal measurements is developing
a nondisruptive and nondestructive technique capable of analyzing
the exogenous compounds contained within the environment. A methodology
using liquid microjunction-surface sampling probe-mass spectrometry
(LMJ-SSP-MS) and microfluidic devices with attached microporous membranes
was developed for in situ, spatiotemporal measurement
of amino acids (AAs) from bacterial biofilms and plant roots. Exuded
arginine was measured from a living Pantoea YR343
biofilm, which resulted in a chemical image indicative of biofilm
growth within the device. Spot sampling along the roots of Populus trichocarpa with the LMJ-SSP-MS resulted in the
detection of 15 AAs. Variation in AA concentrations across the root
system was observed, indicating that exudation is not homogeneous
and may be linked to local rhizosphere architecture and different
biological processes along the root.
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