Microfluidics for Biological Applications
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While beneficial for some samples, this process is not suitable for all experimental conditions, cells, or organisms.
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Indeed, most biological samples cannot be maintained so easily during transport, mammalian cells for example ; therefore, a more ready-to-use approach is needed to expedite collaborative exchanges by overcoming initial barriers to implementing microfluidics in biology. As an alternative to preparing cultures for transport, we propose a ready-to-use approach where microfluidics can be preconditioned dry, with vacuum, or hydrated and conditioned with agar to prevent drying and to provide an immediate-use product for the recipient to inoculate.
From our experience, the best process for loading chambers and successful culture is to initiate microfluidic priming within a 2-day shipping window. Longer shipments or travel may increase the time to fill, or rate of failure for this microfluidic preparation. Another possible explanation includes, age- and environmental-related temperature aging or stiffening of the elastomeric material.
For example, twenty-eight sample chambers were deployed in response to interests communicated at a bacterial-fungal interactions workshop.
Factors influencing the rate of successfully obtained cultures include, travel time, sample handling and storage during transit, and the learning curve for handling and filling microfluidics. Notwithstanding these possible limitations, some users reported that the devices were simple and easy to use, while others needed more replicates to successfully implement microfluidics in their experimental system.
Microfluidics - A Review with Selected Biological Applications I
In this regard, we cannot overlook the procedural requirements of the biological system being integrated into the microfluidic chip. Culturing cells at room temperature without specialized gaseous environmental culture conditions e. By extension, learning to use microfluidics in a biological safety cabinet is more challenging than working with them on a benchtop. Overall, we were pleased with the ability of this packaging processes to accelerate collaborative research and to quickly achieve productive results.
It is our goal that this vacuum-packing process can be adopted and decrease the barriers to implementing microfluidics by providing a process to get microfluidics in-hand to end-users for testing and implementing in biological science. Future research on fungal-bacterial interactions with bacteria could include fluorescent vitality stains for assessing fungi and bacteria, the use of fluorescence reporters for monitoring gene expression activity in microbes, or the use of chamber doors for sampling cells, nucleotides or metabolites from interacting tissues.
Of special interest would be to investigate 1 if the architecture of the chamber can influence the outcome of a biological interaction related to previous studies, 2 how bacteria and fungi enter and grow within plant roots, 3 how environmental conditions influence cytoplasmic flow of cellular contents, or the colonization and community structure of the rhizosphere. Microfluidic platforms have proven to be indispensable tools for probing processes of cellular function, organismal behavior, and environmental interactions. While biomedical disciplines have greatly benefited from the discoveries enabled by microfluidics [ 22 ], botany and mycology are ripe for microfluidic-enabled discoveries and solutions.
Focus areas include, but are not limited to, hyphal chemotropism, fungal pathogenecity, tripartite interactions of the rhizosphere. More specifically, bacterial-fungal interaction studies are also in their infancy, as evidenced by much higher resolution studies [ 70 , 71 , 72 ]. While microfluidics offer the opportunity to engineer microcosms for probing the physical, chemical and biologicals aspects of multispecies interactions, these devices are convenient but not required for such imaging studies in general.
Microfluidics technology: future prospects for molecular diagnostics
Plant-fungal mutualism studies offer ample promise, and plant-bacteria interactions are valuable as well in medicine as for bioenergy, food crop production, and purifying natural resources [ 57 , 58 , 59 , 60 ]. Prior to deploying vacuum-equilibrated microfluidics for studying the biology of branching structures, we developed an architecture that enables low-density, high resolution access to branched hyphae for fungal-bacterial interaction studies.
Spoke-wheel microfluidics for ectomycorrhizal bacterial-fungal interactions were tested in-house before shipping to international colleagues for implementation Fig. This work demonstrates the ability to accelerate studies of defined microcosms within microfluidic platforms by incorporating fabrication and sterilization processes into the packing process for shipping and sharing. We have characterized and deployed an affordable, readily available, and inexpensive process that solves the priming challenge, for new collaborators, by storing sterile dry vacuum-equilibrated microfluidics in a vacuum-sealed pouch for easy fluid priming, anytime, anywhere.
This preparation and packaging process allow the end user to achieve fluid-primed microfluidics without the need for pumping systems.
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The need for fluidic priming at the fabrication source is eliminated, the shipment of fluids is avoided, and the ease of priming microfluidics is achieved. The end user specifies and supplies the fluids, thereby alleviating the supplier from the need to fluid-match recipes, requirements pH , or conditions prevent fluid aging or contamination.
We present a process that accelerates the adoption of microfluidics in labs lacking experience with the technique. The adaptation of challenging microtechnological methods across discipline boundaries can further accelerate the development and implementation of defined microcosms and engineered niches to better resolve fungal, bacterial, and plant interactions. By extension, microfluidics could also allow for the extraction or injection of biological contents for biochemical analyses and genome regulation studies, respectively.
Breaking down scientific barriers through technological simplifications enables studies that have the power to move science forward toward resolving mechanisms of complex and pressing biological problems e. An error occurred during the publication of a number of articles in Fungal Biology and Biotechnology. Several articles were published in volume 6 with a duplicate citation number.
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3D printed microfluidics for biological applications - Lab on a Chip (RSC Publishing)
Mechanistic basis of branch-site selection in filamentous bacteria. PLoS Comput Biol. Regulation of apical growth and hyphal branching in Streptomyces. Curr Opin Microbiol. Decreasing the hyphal branching rate of Saccharopolyspora erythraea NRRL leads to increased resistance to breakage and increased antibiotic production.
Biotechnol Bioeng. Branching in plants. The application of Arabidopsis thaliana in studying tripartite interactions among plants, beneficial fungal endophytes and biotrophic plant-parasitic nematodes. Impact of bacterial-fungal interactions on the colonization of the endosphere. Trends Plant Sci.
Bacterial-fungal interactions: ecology, mechanisms and challenges. Chklovskii DB. Synaptic connectivity and neuronal morphology. Fibrillar amyloid deposition leads to local synaptic abnormalities and breakage of neuronal branches. Nat Neurosci. Microfluidic quantification of yeast surface adhesion. A microfluidic platform with integrated sensing pillars for protrusive force measurements on neurospora crassa.
IEEE; A microfluidic device for massively parallel, whole-lifespan imaging of single fission yeast cells. Microfluidic system for rapid detection of airborne pathogenic fungal spores. ACS Sens. Droplet-based microfluidic high-throughput screening of heterologous enzymes secreted by the yeast Yarrowia lipolytica. Microb Cell Fact.
Various On-Chip Sensors with Microfluidics for Biological Applications
New perspectives on neuronal development via microfluidic environments. Trends Neurosci. Microfluidics structures for probing the dynamic behaviour of filamentous fungi. Microelectron Eng. Fungi use efficient algorithms for the exploration of microfluidic networks. Microfluidic analysis of pressure drop and flow behavior in hypertensive micro vessels.
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