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1. Snapshot polarimetric diffuse-specular separation

   https://doi.org/10.1364/OE.460984  

   Dave, Akshat; Hold-Geoffroy, Yannick; Hašan, Miloš; Sunkavalli, Kalyan; Veeraraghavan, Ashok (September 2022, Optics Express)

   We present a polarization-based approach to perform diffuse-specular separation from a single polarimetric image, acquired using a flexible, practical capture setup. Our key technical insight is that, unlike previous polarization-based separation methods that assume completely unpolarized diffuse reflectance, we use a more general polarimetric model that accounts for partially polarized diffuse reflections. We capture the scene with a polarimetric sensor and produce an initial analytical diffuse-specular separation that we further pass into a deep network trained to refine the separation. We demonstrate that our combination of analytical separation and deep network refinement produces state-of-the-art diffuse-specular separation, which enables image-based appearance editing of dynamic scenes and enhanced appearance estimation. 

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2. Assessing the Discriminatory Capabilities of iEK Devices under DC and DC-Biased AC Stimulation Potentials

   https://doi.org/10.3390/mi14122239  

   Nasir_Ahamed, Nuzhet Nihaar; Mendiola-Escobedo, Carlos A; Perez-Gonzalez, Victor H; Lapizco-Encinas, Blanca H (December 2023, Micromachines)

   There is a rising need for rapid and reliable analytical methods for separating microorganisms in clinical and biomedical applications. Microscale-insulator-based electrokinetic (iEK) systems have proven to be robust platforms for assessing a wide variety of microorganisms. Traditionally, iEK systems are usually stimulated with direct-current (DC) potentials. This work presents a comparison between using DC potentials and using DC-biased alternating-current (AC) potentials in iEK systems for the separation of microorganisms. The present study, which includes mathematical modeling and experimentation, compares the separation of bacterial and yeast cells in two distinct modes by using DC and DC-biased AC potentials. The quality of both separations, assessed in terms of separation resolution (Rs), showed a complete separation (Rs = 1.51) with the application of a DC-biased low-frequency AC signal but an incomplete separation (Rs = 0.55) with the application of an RMS-equivalent DC signal. Good reproducibility between experimental repetitions (<10%) was obtained, and good agreement (~18% deviation) was observed between modeling and experimental retention times. The present study demonstrates the potential of extending the limits of iEK systems by employing DC-biased AC potentials to perform discriminatory separations of microorganisms that are difficult to separate with the application of DC potentials. 

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3. Identifying sequence perturbations to an intrinsically disordered protein that determine its phase-separation behavior

   https://doi.org/10.1073/pnas.2000223117  

   Schuster, Benjamin S.; Dignon, Gregory L.; Tang, Wai Shing; Kelley, Fleurie M.; Ranganath, Aishwarya Kanchi; Jahnke, Craig N.; Simpkins, Alison G.; Regy, Roshan Mammen; Hammer, Daniel A.; Good, Matthew C.; et al (May 2020, Proceedings of the National Academy of Sciences)

   Phase separation of intrinsically disordered proteins (IDPs) commonly underlies the formation of membraneless organelles, which compartmentalize molecules intracellularly in the absence of a lipid membrane. Identifying the protein sequence features responsible for IDP phase separation is critical for understanding physiological roles and pathological consequences of biomolecular condensation, as well as for harnessing phase separation for applications in bioinspired materials design. To expand our knowledge of sequence determinants of IDP phase separation, we characterized variants of the intrinsically disordered RGG domain from LAF-1, a model protein involved in phase separation and a key component of P granules. Based on a predictive coarse-grained IDP model, we identified a region of the RGG domain that has high contact probability and is highly conserved between species; deletion of this region significantly disrupts phase separation in vitro and in vivo. We determined the effects of charge patterning on phase behavior through sequence shuffling. We designed sequences with significantly increased phase separation propensity by shuffling the wild-type sequence, which contains well-mixed charged residues, to increase charge segregation. This result indicates the natural sequence is under negative selection to moderate this mode of interaction. We measured the contributions of tyrosine and arginine residues to phase separation experimentally through mutagenesis studies and computationally through direct interrogation of different modes of interaction using all-atom simulations. Finally, we show that despite these sequence perturbations, the RGG-derived condensates remain liquid-like. Together, these studies advance our fundamental understanding of key biophysical principles and sequence features important to phase separation. 

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4. Numerical study of dielectrophoresis‐modified inertial migration for overlapping sized cell separation

   https://doi.org/10.1002/elps.202100187  

   Khan, Mohammed; Chen, Xiaolin (January 2022, ELECTROPHORESIS)

   Abstract Circulating tumor cells (CTCs) have been proven to have significant prognostic, diagnostic, and clinical values in early‐stage cancer detection and treatment. The efficient separation of CTCs from peripheral blood can ensure intact and viable CTCs and can, thus, give proper genetic characterization and drug innovation. In this study, continuous and high‐throughput separation of MDA‐231 CTCs from overlapping sized white blood cells (WBCs) is achieved by modifying inertial cell focusing with dielectrophoresis (DEP) in a single‐stage microfluidic platform by numeric simulation. The DEP is enabled by embedding interdigitated electrodes with alternating field control on a serpentine microchannel to avoid creating two‐stage separation. Rather than using the electrokinetic migration of cells which slows down the throughput, the system leverages the inertial microfluidic flow to achieve high‐speed continuous separation. The cell migration and cell positioning characteristics are quantified through coupled physics analyses to evaluate the effects of the applied voltages and Reynolds numbers (Re) on the separation performance. The results indicate that the introduction of DEP successfully migrates WBCs away from CTCs and that separation of MDA‐231 CTCs from similar sized WBCs at a highReof 100 can be achieved with a low voltage of magnitude 4 ×10⁶ V/m. Additionally, the viability of MDA‐231 CTCs is expected to be sustained after separation due to the short‐term DEP exposure. The developed technique could be exploited to design active microchips for high‐throughput separation of mixed cell beads despite their significant size overlap, using DEP‐modified inertial focusing controlled simply by adjusting the applied external field. 

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5. Investigation of induced electroosmotic flow in small‐scale capillary electrophoresis devices: Strategies for control and reversal

   https://doi.org/10.1002/elps.202400107  

   De_Silva, Miyuru; Opallage, Prabhavie_M; Dunn, Robert_C (July 2024, ELECTROPHORESIS)

   Abstract Electroosmotic flow (EOF) is the bulk flow of solution in a capillary or microchannel induced by an applied electric potential. For capillary and microchip electrophoresis, the EOF enables analysis of both cations and anions in one separation and can be varied to modify separation speed and resolution. The EOF arises from an electrical double layer at the capillary wall and is normally controlled through the pH and ionic strength of the background buffer or with the use of additives. Understanding and controlling the electrical double layer is therefore critical for maintaining acceptable repeatability during method development. Surprisingly, in fused silica capillaries at low pH, studies observe an EOF even though the capillary surface should be neutralized. Previous work has suggested the presence of an “induced electroosmotic flow” from radial electric fields generated across the capillary wall due to the separation voltage and grounded components external to the capillary. Using thin‐wall (15 µm) fused silica separation capillaries to facilitate the study of radial fields, we show that the EOF mobility depends on both the separation voltage and the location of external grounds. This is consistent with the induced EOF model, in which radial electric fields embed positive charges at the capillary walls to create an electrical double layer. The magnitude of the effect is characterized and shown to have long‐range influences that are difficult to completely null by moving grounded components away from the separation capillary. Instead, active EOF control using externally applied potentials or a passive approach using a negative separation voltage are discussed as two possible methods for controlling the induced EOF. Both methods can reverse the EOF and improve the resolution and peak efficiency in amino acid separations. 

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