@article {68471, title = {Leveraging the model-experiment loop: Examples from cellular slime mold chemotaxis.}, journal = {Exp Cell Res}, volume = {418}, year = {2022}, month = {09/2022}, pages = {113218}, abstract = {

Interplay between models and experimental data advances discovery and understanding in biology, particularly when models generate predictions that allow well-designed experiments to distinguish between alternative mechanisms. To illustrate how this feedback between models and experiments can lead to key insights into biological mechanisms, we explore three examples from cellular slime mold chemotaxis. These examples include studies that identified chemotaxis as the primary mechanism behind slime mold aggregation, discovered that cells likely measure chemoattractant gradients by sensing concentration differences across cell length, and tested the role of cell-associated chemoattractant degradation in shaping chemotactic fields. Although each study used a different model class appropriate to their hypotheses - qualitative, mathematical, or simulation-based - these examples all highlight the utility of modeling to formalize assumptions and generate testable predictions. A central element of this framework is the iterative use of models and experiments, specifically: matching experimental designs to the models, revising models based on mismatches with experimental data, and validating critical model assumptions and predictions with experiments. We advocate for continued use of this interplay between models and experiments to advance biological discovery.

}, keywords = {Chemotactic Factors, Chemotaxis, Computer Simulation, Dictyosteliida, Dictyostelium, Models, Biological}, issn = {1090-2422}, doi = {10.1016/j.yexcr.2022.113218}, author = {Zhu, Xinwen and Hager, Emily R and Huyan, Chuqiao and Sgro, Allyson E} } @article {68472, title = {Microbial models of development: Inspiration for engineering self-assembled synthetic multicellularity.}, journal = {Semin Cell Dev Biol}, year = {2022}, month = {05/2022}, abstract = {

While the field of synthetic developmental biology has traditionally focused on the study of the rich developmental processes seen in metazoan systems, an attractive alternate source of inspiration comes from microbial developmental models. Microbes face unique lifestyle challenges when forming emergent multicellular collectives. As a result, the solutions they employ can inspire the design of novel multicellular systems. In this review, we dissect the strategies employed in multicellular development by two model microbial systems: the cellular slime mold Dictyostelium discoideum and the biofilm-forming bacterium Bacillus subtilis. Both microbes face similar challenges but often have different solutions, both from metazoan systems and from each other, to create emergent multicellularity. These challenges include assembling and sustaining a critical mass of participating individuals to support development, regulating entry into development, and assigning cell fates. The mechanisms these microbial systems exploit to robustly coordinate development under a wide range of conditions offer inspiration for a new toolbox of solutions to the synthetic development community. Additionally, recreating these phenomena synthetically offers a pathway to understanding the key principles underlying how these behaviors are be coordinated naturally.

}, issn = {1096-3634}, doi = {10.1016/j.semcdb.2022.04.014}, author = {Ricci-Tam, Chiara and Kuipa, Sophia and Kostman, Maya Peters and Aronson, Mark S and Sgro, Allyson E} } @article {68475, title = {Arnold tongues in oscillator systems with nonuniform spatial driving.}, journal = {Phys Rev E}, volume = {103}, year = {2021}, month = {04/2021}, pages = {042211}, abstract = {

Nonlinear oscillator systems are ubiquitous in biology and physics, and their control is a practical problem in many experimental systems. Here we study this problem in the context of the two models of spatially coupled oscillators: the complex Ginzburg-Landau equation (CGLE) and a generalization of the CGLE in which oscillators are coupled through an external medium (emCGLE). We focus on external control drives that vary in both space and time. We find that the spatial distribution of the drive signal controls the frequency ranges over which oscillators synchronize to the drive and that boundary conditions strongly influence synchronization to external drives for the CGLE. Our calculations also show that the emCGLE has a low density regime in which a broad range of frequencies can be synchronized for low drive amplitudes. We study the bifurcation structure of these models and find that they are very similar to results for the driven Kuramoto model, a system with no spatial structure. We conclude by discussing qualitative implications of our results for controlling coupled oscillator systems such as the social amoebae Dictyostelium and populations of Belousov Zhabotinsky (BZ) catalytic particles using spatially structured external drives.

}, issn = {2470-0053}, doi = {10.1103/PhysRevE.103.042211}, author = {Golden, Alexander and Sgro, Allyson E and Mehta, Pankaj} } @article {68473, title = {Erratum: Label-free imaging of fibroblast membrane interfaces and protein signatures with vibrational infrared photothermal and phase signals: publisher{\textquoteright}s note.}, journal = {Biomed Opt Express}, volume = {12}, year = {2021}, month = {09/2021}, pages = {5400}, abstract = {

[This corrects the article on p. 303 in vol. 12, PMID: 33520386.].

}, issn = {2156-7085}, doi = {10.1364/BOE.438946}, author = {Samolis, Panagis D and Langley, Daniel and O{\textquoteright}Reilly, Breanna M and Oo, Zay and Hilzenrat, Geva and Erramilli, Shyamsunder and Sgro, Allyson E and McArthur, Sally and Sander, Michelle Y} } @article {68476, title = {Label-free imaging of fibroblast membrane interfaces and protein signatures with vibrational infrared photothermal and phase signals.}, journal = {Biomed Opt Express}, volume = {12}, year = {2021}, month = {12/2020}, pages = {303-319}, abstract = {

Label-free vibrational imaging of biological samples has attracted significant interest due to its integration of structural and chemical information. Vibrational infrared photothermal amplitude and phase signal (VIPPS) imaging provide label-free chemical identification by targeting the characteristic resonances of biological compounds that are present in the mid-infrared fingerprint region (3 {\textmu}m - 12 {\textmu}m). High contrast imaging of subcellular features and chemical identification of protein secondary structures in unlabeled and labeled fibroblast cells embedded in a collagen-rich extracellular matrix is demonstrated by combining contrast from absorption signatures (amplitude signals) with sensitive detection of different heat properties (lock-in phase signals). We present that the detectability of nano-sized cell membranes is enhanced to well below the optical diffraction limit since the membranes are found to act as thermal barriers. VIPPS offers a novel combination of chemical imaging and thermal diffusion characterization that paves the way towards label-free imaging of cell models and tissues as well as the study of intracellular heat dynamics.

}, issn = {2156-7085}, doi = {10.1364/BOE.411888}, author = {Samolis, Panagis D and Langley, Daniel and O{\textquoteright}Reilly, Breanna M and Oo, Zay and Hilzenrat, Geva and Erramilli, Shyamsunder and Sgro, Allyson E and McArthur, Sally and Sander, Michelle Y} } @article {68474, title = {Ventral stress fibers induce plasma membrane deformation in human fibroblasts.}, journal = {Mol Biol Cell}, volume = {32}, year = {2021}, month = {08/2021}, pages = {1707-1723}, abstract = {

Interactions between the actin cytoskeleton and the plasma membrane are important in many eukaryotic cellular processes. During these processes, actin structures deform the cell membrane outward by applying forces parallel to the fiber{\textquoteright}s major axis (as in migration) or they deform the membrane inward by applying forces perpendicular to the fiber{\textquoteright}s major axis (as in the contractile ring during cytokinesis). Here we describe a novel actin-membrane interaction in human dermal myofibroblasts. When labeled with a cytosolic fluorophore, the myofibroblasts displayed prominent fluorescent structures on the ventral side of the cell. These structures are present in the cell membrane and colocalize with ventral actin stress fibers, suggesting that the stress fibers bend the membrane to form a "cytosolic pocket" that the fluorophores diffuse into, creating the observed structures. The existence of this pocket was confirmed by transmission electron microscopy. While dissolving the stress fibers, inhibiting fiber protein binding, or inhibiting myosin II binding of actin removed the observed pockets, modulating cellular contractility did not remove them. Taken together, our results illustrate a novel actin-membrane bending topology where the membrane is deformed outward rather than being pinched inward, resembling the topological inverse of the contractile ring found in cytokinesis.

}, keywords = {Actins, Cell Membrane, Cells, Cultured, Cytochalasin D, Cytosol, Fibroblasts, Fluorescent Dyes, Humans, Infant, Myosin Type II, Ouabain, Skin, Stress Fibers, Transforming Growth Factor beta1}, issn = {1939-4586}, doi = {10.1091/mbc.E21-03-0096}, author = {Ghilardi, Samuel J and Aronson, Mark S and Sgro, Allyson E} } @article {68477, title = {Eco-evolutionary significance of "loners".}, journal = {PLoS Biol}, volume = {18}, year = {2020}, month = {03/2020}, pages = {e3000642}, abstract = {

Loners-individuals out of sync with a coordinated majority-occur frequently in nature. Are loners incidental byproducts of large-scale coordination attempts, or are they part of a mosaic of life-history strategies? Here, we provide empirical evidence of naturally occurring heritable variation in loner behavior in the model social amoeba Dictyostelium discoideum. We propose that Dictyostelium loners-cells that do not join the multicellular life stage-arise from a dynamic population-partitioning process, the result of each cell making a stochastic, signal-based decision. We find evidence that this imperfectly synchronized multicellular development is affected by both abiotic (environmental porosity) and biotic (signaling) factors. Finally, we predict theoretically that when a pair of strains differing in their partitioning behavior coaggregate, cross-signaling impacts slime-mold diversity across spatiotemporal scales. Our findings suggest that loners could be critical to understanding collective and social behaviors, multicellular development, and ecological dynamics in D. discoideum. More broadly, across taxa, imperfect coordination of collective behaviors might be adaptive by enabling diversification of life-history strategies.

}, keywords = {Biological Evolution, Dictyostelium, Models, Biological, Quorum Sensing, Spatio-Temporal Analysis, Stochastic Processes}, issn = {1545-7885}, doi = {10.1371/journal.pbio.3000642}, author = {Rossine, Fernando W and Martinez-Garcia, Ricardo and Sgro, Allyson E and Gregor, Thomas and Tarnita, Corina E} } @article {68478, title = {Intracellular signaling dynamics and their role in coordinating tissue repair.}, journal = {Wiley Interdiscip Rev Syst Biol Med}, volume = {12}, year = {2020}, month = {05/2020}, pages = {e1479}, abstract = {

Tissue repair is a complex process that requires effective communication and coordination between cells across multiple tissues and organ systems. Two of the initial intracellular signals that encode injury signals and initiate tissue repair responses are calcium and extracellular signal-regulated kinase (ERK). However, calcium and ERK signaling control a variety of cellular behaviors important for injury repair including cellular motility, contractility, and proliferation, as well as the activity of several different transcription factors, making it challenging to relate specific injury signals to their respective repair programs. This knowledge gap ultimately hinders the development of new wound healing therapies that could take advantage of native cellular signaling programs to more effectively repair tissue damage. The objective of this review is to highlight the roles of calcium and ERK signaling dynamics as mechanisms that link specific injury signals to specific cellular repair programs during epithelial and stromal injury repair. We detail how the signaling networks controlling calcium and ERK can now also be dissected using classical signal processing techniques with the advent of new biosensors and optogenetic signal controllers. Finally, we advocate the importance of recognizing calcium and ERK dynamics as key links between injury detection and injury repair programs that both organize and execute a coordinated tissue repair response between cells across different tissues and organs. This article is categorized under: Models of Systems Properties and Processes \> Mechanistic Models Biological Mechanisms \> Cell Signaling Laboratory Methods and Technologies \> Imaging Models of Systems Properties and Processes \> Organ, Tissue, and Physiological Models.

}, keywords = {Animals, Calcium, Cell Movement, Epithelial Cells, Extracellular Signal-Regulated MAP Kinases, Humans, Models, Biological, NF-kappa B, Signal Transduction, Wound Healing}, issn = {1939-005X}, doi = {10.1002/wsbm.1479}, author = {Ghilardi, Samuel J and O{\textquoteright}Reilly, Breanna M and Sgro, Allyson E} } @article {68482, title = {Fitness tradeoffs between spores and nonaggregating cells can explain the coexistence of diverse genotypes in cellular slime molds.}, journal = {Proc Natl Acad Sci U S A}, volume = {112}, year = {2015}, month = {03/2015}, pages = {2776-81}, abstract = {

Cellular slime molds, including the well-studied Dictyostelium discoideum, are amoebae whose life cycle includes both a single-cellular and a multicellular stage. To achieve the multicellular stage, individual amoebae aggregate upon starvation to form a fruiting body made of dead stalk cells and reproductive spores, a process that has been described in terms of cooperation and altruism. When amoebae aggregate they do not perfectly discriminate against nonkin, leading to chimeric fruiting bodies. Within chimeras, complex interactions among genotypes have been documented, which should theoretically reduce genetic diversity. This is however inconsistent with the great diversity of genotypes found in nature. Recent work has shown that a little-studied component of D. discoideum fitness--the loner cells that do not participate in the aggregation--can be selected for depending on environmental conditions and that, together with the spores, they could represent a bet-hedging strategy. We suggest that in all cellular slime molds the existence of loners could resolve the apparent diversity paradox in two ways. First, if loners are accounted for, then apparent genotypic skew in the spores of chimeras could simply be the result of different investments into spores versus loners. Second, in an ecosystem with multiple local environments differing in their food recovery characteristics and connected globally via weak-to-moderate dispersal, coexistence of multiple genotypes can occur. Finally, we argue that the loners make it impossible to define altruistic behavior, winners or losers, without a clear description of the ecology.

}, keywords = {Dictyostelium, Ecosystem, Life Cycle Stages, Spores, Protozoan}, issn = {1091-6490}, doi = {10.1073/pnas.1424242112}, author = {Tarnita, Corina E and Washburne, Alex and Martinez-Garcia, Ricardo and Sgro, Allyson E and Levin, Simon A} } @article {68481, title = {From intracellular signaling to population oscillations: bridging size- and time-scales in collective behavior.}, journal = {Mol Syst Biol}, volume = {11}, year = {2015}, month = {01/2015}, pages = {779}, abstract = {

Collective behavior in cellular populations is coordinated by biochemical signaling networks within individual cells. Connecting the dynamics of these intracellular networks to the population phenomena they control poses a considerable challenge because of network complexity and our limited knowledge of kinetic parameters. However, from physical systems, we know that behavioral changes in the individual constituents of a collectively behaving system occur in a limited number of well-defined classes, and these can be described using simple models. Here, we apply such an approach to the emergence of collective oscillations in cellular populations of the social amoeba Dictyostelium discoideum. Through direct tests of our model with quantitative in vivo measurements of single-cell and population signaling dynamics, we show how a simple model can effectively describe a complex molecular signaling network at multiple size and temporal scales. The model predicts novel noise-driven single-cell and population-level signaling phenomena that we then experimentally observe. Our results suggest that like physical systems, collective behavior in biology may be universal and described using simple mathematical models.

}, keywords = {Dictyostelium, Gene Expression Regulation, Image Processing, Computer-Assisted, Microfluidic Analytical Techniques, Molecular Dynamics Simulation, Signal Transduction}, issn = {1744-4292}, doi = {10.15252/msb.20145352}, author = {Sgro, Allyson E and Schwab, David J and Noorbakhsh, Javad and Mestler, Troy and Mehta, Pankaj and Gregor, Thomas} } @article {68480, title = {Modeling oscillations and spiral waves in Dictyostelium populations.}, journal = {Phys Rev E Stat Nonlin Soft Matter Phys}, volume = {91}, year = {2015}, month = {06/2015}, pages = {062711}, abstract = {

Unicellular organisms exhibit elaborate collective behaviors in response to environmental cues. These behaviors are controlled by complex biochemical networks within individual cells and coordinated through cell-to-cell communication. Describing these behaviors requires new mathematical models that can bridge scales-from biochemical networks within individual cells to spatially structured cellular populations. Here we present a family of "multiscale" models for the emergence of spiral waves in the social amoeba Dictyostelium discoideum. Our models exploit new experimental advances that allow for the direct measurement and manipulation of the small signaling molecule cyclic adenosine monophosphate (cAMP) used by Dictyostelium cells to coordinate behavior in cellular populations. Inspired by recent experiments, we model the Dictyostelium signaling network as an excitable system coupled to various preprocessing modules. We use this family of models to study spatially unstructured populations of "fixed" cells by constructing phase diagrams that relate the properties of population-level oscillations to parameters in the underlying biochemical network. We then briefly discuss an extension of our model that includes spatial structure and show how this naturally gives rise to spiral waves. Our models exhibit a wide range of novel phenomena. including a density-dependent frequency change, bistability, and dynamic death due to slow cAMP dynamics. Our modeling approach provides a powerful tool for bridging scales in modeling of Dictyostelium populations.

}, keywords = {Cyclic AMP, Dictyostelium, Extracellular Space, Kinetics, Models, Biological, Signal Transduction}, issn = {1550-2376}, doi = {10.1103/PhysRevE.91.062711}, author = {Noorbakhsh, Javad and Schwab, David J and Sgro, Allyson E and Gregor, Thomas and Mehta, Pankaj} } @article {68483, title = {Quantitative biology: where modern biology meets physical sciences.}, journal = {Mol Biol Cell}, volume = {25}, year = {2014}, month = {11/2014}, pages = {3482-5}, abstract = {

Quantitative methods and approaches have been playing an increasingly important role in cell biology in recent years. They involve making accurate measurements to test a predefined hypothesis in order to compare experimental data with predictions generated by theoretical models, an approach that has benefited physicists for decades. Building quantitative models in experimental biology not only has led to discoveries of counterintuitive phenomena but has also opened up novel research directions. To make the biological sciences more quantitative, we believe a two-pronged approach needs to be taken. First, graduate training needs to be revamped to ensure biology students are adequately trained in physical and mathematical sciences and vice versa. Second, students of both the biological and the physical sciences need to be provided adequate opportunities for hands-on engagement with the methods and approaches necessary to be able to work at the intersection of the biological and physical sciences. We present the annual Physiology Course organized at the Marine Biological Laboratory (Woods Hole, MA) as a case study for a hands-on training program that gives young scientists the opportunity not only to acquire the tools of quantitative biology but also to develop the necessary thought processes that will enable them to bridge the gap between these disciplines.

}, keywords = {Cell Biology, Computer Simulation, Education, Graduate, Humans, Models, Biological, Natural Science Disciplines, Students}, issn = {1939-4586}, doi = {10.1091/mbc.E14-08-1286}, author = {Shekhar, Shashank and Zhu, Lian and Mazutis, Linas and Sgro, Allyson E and Fai, Thomas G and Podolski, Marija} } @article {68484, title = {Single-axonal organelle analysis method reveals new protein-motor associations.}, journal = {ACS Chem Neurosci}, volume = {4}, year = {2013}, month = {02/2013}, pages = {277-84}, abstract = {

Axonal transport of synaptic vesicle proteins is required to maintain neurons{\textquoteright} ability to communicate via synaptic transmission. Neurotransmitter-containing synaptic vesicles are assembled at synaptic terminals via highly regulated endocytosis of membrane proteins. These synaptic vesicle membrane proteins are synthesized in the cell body and transported to synapses in carrier vesicles that make their way down axons via microtubule-based transport utilizing kinesin molecular motors. Identifying the cargos that each kinesin motor protein carries from the cell bodies to the synapse is key to understanding both diseases caused by motor protein dysfunction and how synaptic vesicles are assembled. However, obtaining a bulk sample of axonal transport complexes from central nervous system (CNS) neurons to use for identification of their contents has posed a challenge to researchers. To obtain axonal carrier vesicles from primary cultured neurons, we fabricated a microfluidic chip designed to physically isolate axons from dendrites and cell bodies and developed a method to remove bulk axonal samples and label their contents. Synaptic vesicle protein carrier vesicles in these samples were labeled with antibodies to the synaptic vesicle proteins p38, SV2A, and VAMP2, and the anterograde axonal transport motor KIF1A, after which antibody overlap was evaluated using single-organelle TIRF microscopy. This work confirms a previously discovered association between KIF1A and p38 and shows that KIF1A also transports SV2A- and VAMP2-containing carrier vesicles.

}, keywords = {Animals, Axonal Transport, Cells, Cultured, Fluorescent Antibody Technique, Hippocampus, Kinesins, Membrane Glycoproteins, Mice, Microfluidic Analytical Techniques, Microscopy, Confocal, Nerve Tissue Proteins, Neurons, Organelles, Synaptic Vesicles, Synaptophysin, Transport Vesicles, Vesicle-Associated Membrane Protein 2, Vesicular Transport Proteins}, issn = {1948-7193}, doi = {10.1021/cn300136y}, author = {Sgro, Allyson E and Bajjalieh, Sandra M and Chiu, Daniel T} } @article {68485, title = {SnapShot: Optical control and imaging of brain activity.}, journal = {Cell}, volume = {149}, year = {2012}, month = {06/2012}, pages = {1650-1650.e2}, abstract = {

No abstract available.

}, keywords = {Animals, Brain, Microscopy, Fluorescence, Molecular Imaging, Optics and Photonics}, issn = {1097-4172}, doi = {10.1016/j.cell.2012.06.009}, author = {Richard Sun, X and Giovannucci, Andrea and Sgro, Allyson E and Wang, Samuel S-H} } @article {68486, title = {A high-throughput method for generating uniform microislands for autaptic neuronal cultures.}, journal = {J Neurosci Methods}, volume = {198}, year = {2011}, month = {06/2011}, pages = {230-5}, abstract = {

Generating microislands of culture substrate on coverslips by spray application of poly-d lysine is a commonly used method for culturing isolated neurons that form self (autaptic) synapses. This preparation has multiple advantages for studying synaptic transmission in isolation; however, generating microislands by spraying produces islands of non-uniform size and thus cultures vary widely in the number of islands containing single neurons. To address these problems, we developed a high-throughput method for reliably generating uniformly shaped microislands of culture substrate. Stamp molds formed of poly(dimethylsiloxane) (PDMS) were fabricated with arrays of circles and used to generate stamps made of 9.2\% agarose. The agarose stamps were capable of loading sufficient poly D-lysine and collagen dissolved in acetic acid to rapidly generate coverslips containing at least 64 microislands per coverslip. When hippocampal neurons were cultured on these coverslips, there were significantly more single-neuron islands per coverslip. We noted that single neurons tended to form one of three distinct neurite-arbor morphologies, which varied with island size and the location of the cell body on the island. To our surprise, the number of synapses per autaptic neuron did not correlate with arbor shape or island size, suggesting that other factors regulate the number of synapses formed by isolated neurons. The stamping method we report can be used to increase the number of single-neuron islands per culture and aid in the rapid visualization of microislands.

}, keywords = {Animals, Cell Communication, Cell Culture Techniques, Cells, Cultured, Dendrites, Hippocampus, Mice, Neurons, Synapses, Synaptic Transmission}, issn = {1872-678X}, doi = {10.1016/j.jneumeth.2011.04.012}, author = {Sgro, Allyson E and Nowak, Amy L and Austin, Naola S and Custer, Kenneth L and Allen, Peter B and Chiu, Daniel T and Bajjalieh, Sandra M} } @article {4927, title = {Synaptosomes as a platform for loading nanoparticles into synaptic vesicles.}, journal = {ACS Chemical Neuroscience}, volume = {2}, year = {2011}, month = {2011 May 18}, pages = {236-241}, abstract = {

Synaptosomes are intact, isolated nerve terminals that contain the necessary machinery to recycle synaptic vesicles via endocytosis and exocytosis upon stimulation. Here we use this property of synaptosomes to load quantum dots into synaptic vesicles. Vesicles are then isolated from the synaptosomes, providing a method to probe isolated, individual synaptic vesicles where each vesicle contains a single, encapsulated nanoparticle. This technique provided an encapsulation efficiency of \ 16\%, that is, \ 16\% of the vesicles contained a single quantum dot while the remaining vesicles were empty. The ability to load single nanoparticles into synaptic vesicles opens new opportunity for employing various nanoparticle-based sensors to study the dynamics of vesicular transporters.

}, issn = {1948-7193}, doi = {10.1021/cn200009n}, author = {Budzinski, Kristi L and Sgro, Allyson E and Fujimoto, Bryant S and Gadd, Jennifer C and Shuart, Noah G and Gonen, Tamir and Bajjaleih, Sandra M and Chiu, Daniel T} } @article {68487, title = {Droplet freezing, docking, and the exchange of immiscible phase and surfactant around frozen droplets.}, journal = {Lab Chip}, volume = {10}, year = {2010}, month = {07/2010}, pages = {1873-7}, abstract = {

This paper describes a platform for cooling microfluidic chips so as to freeze aqueous droplets flowing in oil. Using a whole-chip cooling chamber, we can control the ambient temperature surrounding a microfluidic chip and induce cooling and freezing inside the channels. When combined with a droplet generation and droplet docking chip, this platform allows for the facile freezing of droplets immobilized in resistance-based docks. Depending on the design and shape of the docks, the frozen droplets can either be trapped stably in the docks or be released because deformed non-frozen aqueous droplets turn spherical when frozen, and thus can become dislodged from the docks. Additionally, using this chamber and chip combination we are able to exchange immiscible phases and surfactants surrounding the frozen droplets. The materials and methods are inexpensive and easily accessible to microfluidics researchers, making this a simple addition to an existing microfluidic platform.

}, keywords = {Equipment Design, Equipment Failure Analysis, Freezing, Ice, Microfluidic Analytical Techniques, Phase Transition, Surface-Active Agents}, issn = {1473-0197}, doi = {10.1039/c001108h}, author = {Sgro, Allyson E and Chiu, Daniel T} } @article {68488, title = {Single-synapse ablation and long-term imaging in live C. elegans.}, journal = {J Neurosci Methods}, volume = {173}, year = {2008}, month = {09/2008}, pages = {20-6}, abstract = {

Synapses are individually operated, computational units for neural communication. To manipulate physically individual synapses in a living organism, we have developed a laser ablation technique for removing single synapses in live neurons in C. elegans that operates without apparent damage to the axon. As a complementary technique, we applied microfluidic immobilization of C. elegans to facilitate long-term fluorescence imaging and observation of neuronal development. With this technique, we directly demonstrated the existence of competition between developing synapses in the HSNL motor neuron.

}, keywords = {Animals, Axons, Behavior, Animal, Caenorhabditis elegans, Diagnostic Imaging, Laser Therapy, Microfluidic Analytical Techniques, Movement, Neurons, Synapses, Time Factors}, issn = {0165-0270}, doi = {10.1016/j.jneumeth.2008.05.007}, author = {Allen, Peter B and Sgro, Allyson E and Chao, Daniel L and Doepker, Byron E and Scott Edgar, J and Shen, Kang and Chiu, Daniel T} } @article {68489, title = {Thermoelectric manipulation of aqueous droplets in microfluidic devices.}, journal = {Anal Chem}, volume = {79}, year = {2007}, month = {07/2007}, pages = {4845-51}, abstract = {

This article describes a method for manipulating the temperature inside aqueous droplets, utilizing a thermoelectric cooler to control the temperature of select portions of a microfluidic chip. To illustrate the adaptability of this approach, we have generated an "ice valve" to stop fluid flow in a microchannel. By taking advantage of the vastly different freezing points for aqueous solutions and immiscible oils, we froze a stream of aqueous droplets that were formed on-chip. By integrating this technique with cell encapsulation into aqueous droplets, we were also able to freeze single cells encased in flowing droplets. Using a live-dead stain, we confirmed the viability of cells was not adversely affected by the process of freezing in aqueous droplets provided cryoprotectants were utilized. When combined with current droplet methodologies, this technology has the potential to both selectively heat and cool portions of a chip for a variety of droplet-related applications, such as freezing, temperature cycling, sample archiving, and controlling reaction kinetics.

}, keywords = {Cell Survival, Freezing, Kinetics, Lymphoma, B-Cell, Microfluidic Analytical Techniques, Mineral Oil, Solutions, Water}, issn = {0003-2700}, doi = {10.1021/ac062458a}, author = {Sgro, Allyson E and Allen, Peter B and Chiu, Daniel T} }