26 research outputs found
Recommended from our members
Modular microporous hydrogels formed from microgel beads with orthogonal thermo-chemical responsivity: Microfluidic fabrication and characterization.
Despite the significant advances in designing injectable bulk hydrogels, the inability to control the pore interconnectivity and decoupling it from the matrix stiffness has tremendously limited the applicability of stiff, flowable hydrogels for 3D cellular engineering, e.g., in hard tissue engineering. To overcome this persistent challenge, here, we introduce a universal method to convert thermosensitive macromolecules with chemically-crosslinkable moieties into annealable building blocks, forming 3D microporous beaded scaffolds in a bottom-up approach. In particular, we show gelatin methacryloyl (GelMA), a widely used biomaterial in tissue engineering, may be converted into physically-crosslinked microbeads using a facile microfluidic approach, followed by flow of the microbead suspension and chemical crosslinking in situ to fabricate microporous beaded GelMA (B-GelMA) scaffolds with interconnected pores, promoting cell functionality and rapid (within minutes) 3D seeding in stiff scaffolds, which are otherwise impossible in the bulk gel counterparts. This novel approach may set the stage for the next generation modular hydrogels with orthogonal porosity and stiffness made up of a broad range of natural and synthetic biomaterials. •This method combines well-known flow focusing microfluidic devices with facile post-processing steps to fabricate microporous scaffolds.•Temperature-driven physical crosslinking of the microbeads enables the facile purification of gel building blocks without further chemical reactions.•This method provides a simple approach to fabricate microporous scaffolds, which overcomes some of the challenges of newly emerging beaded scaffolds, including oxygen-mediated impaired crosslinking
Chimpanzees (Pan troglodytes) do not develop contingent reciprocity in an experimental task
Chimpanzees provide help to unrelated individuals in a broad range of situations. The pattern of helping within pairs suggests that contingent reciprocity may have been an important mechanism in the evolution of altruism in chimpanzees. However, correlational analyses of the cumulative pattern of interactions over time do not demonstrate that helping is contingent upon previous acts of altruism, as required by the theory of reciprocal altruism. Experimental studies provide a controlled approach to examine the importance of contingency in helping interactions. In this study, we evaluated whether chimpanzees would be more likely to provide food to a social partner from their home group if their partner had previously provided food for them. The chimpanzees manipulated a barpull apparatus in which actors could deliver rewards either to themselves and their partners or only to themselves. Our findings indicate that the chimpanzees’ responses were not consistently influenced by the behavior of their partners in previous rounds. Only one of the 11 dyads that we tested demonstrated positive reciprocity. We conclude that contingent reciprocity does not spontaneously arise in experimental settings, despite the fact that patterns of behavior in the field indicate that individuals cooperate preferentially with reciprocating partners
Recommended from our members
Probing the Ultimate Limits of Biology: Developing Microparticle Platforms for High-Throughput Single-Cell Assays
Techniques to analyze and sort single cells based on complex phenotypes such as secreted products have the potential to transform our understanding of cellular biology as well as accelerate the development of next generation cell and antibody therapies. Microfluidic techniques have emerged over the last two decades that enable such assays by creating small compartments that can isolate individual cells. Despite these technical advances, adoption of this technology has been slow due to the complexity of the approaches and the requirement of specialty instruments. To address this issue, we developed a microparticle based approach to create uniform sub-nanoliter water in oil compartments using only standard lab equipment. We refer to these droplets formed with microparticles, dropicles. In this dissertation I will provide an overview of various strategies used to create microscale compartments, specifically to perform single-cell secretion assays. I will then discuss in detail an approach to fabricate and use 3D structured microparticles to perform single-cell secretion assays massively in parallel with standard lab equipment. By making this microparticle approach compatible with existing high-throughput flow cytometers we are able to analyze and sort of over 100,000 single cells based on their secreted products. Lastly to create a truly democratized platform there is a need to be able to manufacture these particles in a scalable manner. In these last chapters I will discuss approaches to fabricate microparticles with different morphologies in high-throughput
Recommended from our members
Probing the Ultimate Limits of Biology: Developing Microparticle Platforms for High-Throughput Single-Cell Assays
Techniques to analyze and sort single cells based on complex phenotypes such as secreted products have the potential to transform our understanding of cellular biology as well as accelerate the development of next generation cell and antibody therapies. Microfluidic techniques have emerged over the last two decades that enable such assays by creating small compartments that can isolate individual cells. Despite these technical advances, adoption of this technology has been slow due to the complexity of the approaches and the requirement of specialty instruments. To address this issue, we developed a microparticle based approach to create uniform sub-nanoliter water in oil compartments using only standard lab equipment. We refer to these droplets formed with microparticles, dropicles. In this dissertation I will provide an overview of various strategies used to create microscale compartments, specifically to perform single-cell secretion assays. I will then discuss in detail an approach to fabricate and use 3D structured microparticles to perform single-cell secretion assays massively in parallel with standard lab equipment. By making this microparticle approach compatible with existing high-throughput flow cytometers we are able to analyze and sort of over 100,000 single cells based on their secreted products. Lastly to create a truly democratized platform there is a need to be able to manufacture these particles in a scalable manner. In these last chapters I will discuss approaches to fabricate microparticles with different morphologies in high-throughput
Recommended from our members
Surface energy minimizing configurations for axisymmetric microparticles
Recommended from our members
Scalable Fabrication and Use of 3D Structured Microparticles Spatially Functionalized with Biomolecules
Microparticles with defined shapes and spatial chemical modification can interface with cells and tissues at the cellular scale. However, conventional methods to fabricate shaped microparticles have trade-offs between the throughput of manufacture and the precision of particle shape and chemical functionalization. Here, we achieved scalable production of hydrogel microparticles at rates of greater than 40 million/hour with localized surface chemistry using a parallelized step emulsification device and temperature-induced phase-separation. The approach harnesses a polymerizable polyethylene glycol (PEG) and gelatin aqueous two-phase system (ATPS) which conditionally phase separates within microfluidically generated droplets. Following droplet formation, phase separation is induced and phase separated droplets are subsequently cross-linked to form uniform crescent and hollow shell particles with gelatin functionalization on the boundary of the cavity. The gelatin localization enabled deterministic cell loading in subnanoliter-sized crescent-shaped particles, which we refer to as nanovials, with cavity dimensions tuned to the size of cells. Loading on nanovials also imparted improved cell viability during analysis and sorting using standard fluorescence activated cell sorters, presumably by protecting cells from shear stress. This localization effect was further exploited to selectively functionalize capture antibodies to nanovial cavities enabling single-cell secretion assays with reduced cross-talk in a simplified format
Numerical investigation of micro- and nanochannel deformation due to discontinuous electroosmotic flow
Large pressures can induce detrimental deformation in micro- and nanofluidic channels. Although this has been extensively studied for systems driven by pressure and/or capillary forces, deflection in electrokinetic systems due to internal pressure gradients caused by non-uniform electric fields has not been widely explored. For example, applying an axial electric field in a channel with a step change in conductivity and/or surface charge can lead to internally generated pressures large enough to cause cavitation, debonding, and/or channel collapse. Finite electric double layers within nanofluidic channels can further complicate the physics involved in the deformation process. In order to design devices and experimental procedures that avoid issues resulting from such deformation, it is imperative to be able to predict deformation for given system parameters. In this work, we numerically investigate pressures resulting from a step change in conductivity and/or surface charge in micro- and nanofluidic channels with both thin and thick double layers. We show an explicit relation of pressure dependence on concentration ratio and electric double layer thickness. Furthermore, we develop a numerical model to predict deformation in such systems and use the model to unearth trends in deformation for various electric double layer thicknesses and both glass and PDMS on glass channels. Our work is particularly impactful for the development and design of micro- and nanofluidic-based devices with gradients in surface charge and/or conductivity, fundamental study of electrokinetic-based cavitation, and other systems that exploit non-uniform electric fields
Quantitative Characterization of the Colloidal Stability of Metallic Nanoparticles Using UV–vis Absorbance Spectroscopy
Plasmonic nanoparticles are used
in a wide variety of applications
over a broad array of fields including medicine, energy, and environmental
chemistry. The continued successful development of this material class
requires the accurate characterization of nanoparticle stability for
a variety of solution-based conditions. Although many characterization
methods exists, there is an absence of a unified, quantitative means
for assessing the colloidal stability of plasmonic nanoparticles.
We present the particle instability parameter (PIP) as a robust, quantitative,
and generalizable characterization technique based on UV–vis
absorbance spectroscopy to characterize colloidal instability. We
validate PIP performance with both traditional and alternative characterization
methods by measuring gold nanorod instability in response to different
salt (NaCl) concentrations. We further measure gold nanorod stability
as a function of solution pH, salt, and buffer (type and concentration),
nanoparticle concentration, and concentration of free surfactant.
Finally, these results are contextualized within the literature on
gold nanorod stability to establish a standardized methodology for
colloidal instability assessment
Recommended from our members
Modular microporous hydrogels formed from microgel beads with orthogonal thermo-chemical responsivity: Microfluidic fabrication and characterization.
Despite the significant advances in designing injectable bulk hydrogels, the inability to control the pore interconnectivity and decoupling it from the matrix stiffness has tremendously limited the applicability of stiff, flowable hydrogels for 3D cellular engineering, e.g., in hard tissue engineering. To overcome this persistent challenge, here, we introduce a universal method to convert thermosensitive macromolecules with chemically-crosslinkable moieties into annealable building blocks, forming 3D microporous beaded scaffolds in a bottom-up approach. In particular, we show gelatin methacryloyl (GelMA), a widely used biomaterial in tissue engineering, may be converted into physically-crosslinked microbeads using a facile microfluidic approach, followed by flow of the microbead suspension and chemical crosslinking in situ to fabricate microporous beaded GelMA (B-GelMA) scaffolds with interconnected pores, promoting cell functionality and rapid (within minutes) 3D seeding in stiff scaffolds, which are otherwise impossible in the bulk gel counterparts. This novel approach may set the stage for the next generation modular hydrogels with orthogonal porosity and stiffness made up of a broad range of natural and synthetic biomaterials. •This method combines well-known flow focusing microfluidic devices with facile post-processing steps to fabricate microporous scaffolds.•Temperature-driven physical crosslinking of the microbeads enables the facile purification of gel building blocks without further chemical reactions.•This method provides a simple approach to fabricate microporous scaffolds, which overcomes some of the challenges of newly emerging beaded scaffolds, including oxygen-mediated impaired crosslinking