Search for: All records

This Tutorial Review highlights strategies for leveraging the micron-to-submicron-scale additive manufacturing technique, “direct laser writing”, to enable 3D microfluidic technologies.

 

Broadband radio waves emitted from pulsars are distorted and delayed as they propagate toward the Earth due to interactions with the free electrons that compose the interstellar medium (ISM), with lower radio frequencies being more impacted than higher frequencies. Multipath propagation in the ISM results in both later times of arrival for the lower frequencies and causes the observed pulse to arrive with a broadened tail described via the pulse broadening function. We employ the CLEAN deconvolution technique to recover the pulse broadening timescale and by proxy the intrinsic pulse shape. This work expands upon previous descriptions of CLEAN deconvolution used in pulse broadening analyses by parameterizing the efficacy on simulated data and developing a suite of tests to establish which of a set of figures of merit leads to an automatic and consistent determination of the scattering timescale and its uncertainty. We compare our algorithm to the cyclic spectroscopy method of estimating the scattering timescale, specifically to the simulations performed in Dolch et al. (2021). We test our improved algorithm on the highly scattered millisecond pulsar J1903+0327, showing the scattering timescale to change over years, consistent with estimates of the refractive timescale of the pulsar.

 

Multi-stage fluidic reaction schemes for suspended particles (e.g., micro/nanospheres, cells, bacterial species, and extracellular vesicles) underly a diversity of chemical and biological applications. Conventional methods for executing such protocols can be exceedingly time, labor, and/or cost intensive. Microfluidic strategies can address these drawbacks; however, such technologies typically rely on clean room-based microfabrication that suffer from similar deficits for manufacturing the chips. To simultaneously overcome these challenges, here we explore the use of the submicron-scale additive manufacturing approach, “Two-Photon Direct Laser Writing (DLW)”, as a means for fabricating micro-fluidic “Deterministic Lateral Displacement (DLD)” arrays capable of passively guiding suspended particles across discrete, adjacent flow streams—the fundamental capability of continuous-flow multi-stage particle microreactors. Experimental results from microfluidic experimentation with 5 μm-in-diameter fluorescent particles revealed effective particle transport across flow streams, with 87.5% of fluorescent peaks detected in the designated, opposing outlet following the DLD array. These results suggest utility of the presented approach for micro- and nanoparticle-based microfluidic reactors targeting wide-ranging chemical and biological applications. 

The ability to reconstitute natural glycosylation pathways or prototype entirely new ones from scratch is hampered by the limited availability of functional glycoenzymes, many of which are membrane proteins that fail to express in heterologous hosts. Here, we describe a strategy for topologically converting membrane-bound glycosyltransferases (GTs) into water soluble biocatalysts, which are expressed at high levels in the cytoplasm of living cells with retention of biological activity. We demonstrate the universality of the approach through facile production of 98 difficult-to-express GTs, predominantly of human origin, across several commonly used expression platforms. Using a subset of these water-soluble enzymes, we perform structural remodeling of both free and protein-linked glycans including those found on the monoclonal antibody therapeutic trastuzumab. Overall, our strategy for rationally redesigning GTs provides an effective and versatile biosynthetic route to large quantities of diverse, enzymatically active GTs, which should find use in structure-function studies as well as in biochemical and biomedical applications involving complex glycomolecules.

 

Deterministic lateral displacement (DLD) is a microfluidic micro/nanopost array-based technique for size-based particle separations. A key challenge in scaling DLD for handling smaller particles is that creating such “nanoDLD” arrays can be cost-intensive with substantial technical hurdles. To circumvent such issues, here we explore a new “hexagonally arranged triangles (HAT)” DLD geometry that is based on patterns associated with nanosphere lithography (NSL). Finite element simulations and preliminary experiments with 0.86 μm and 4.7 μm particles suggest effective separation capabilities of the HAT-DLD approach, marking an important first step toward new classes of nanoDLD arrays fabricated through bottom-up, self-assembly-based NSL.