Direct Measurements of Colloidal Solvophoresis under Imposed Solvent and Solute Gradients

We describe a microfluidic system that enables direct visualization and measurement of diffusiophoretic migration of colloids in response to imposed solution gradients. Such measurements have proven difficult or impossible in macroscopic systems due to difficulties in establishing solution gradients that are sufficiently strong yet hydrodynamically stable. We validate the system with measurements of the concentration-dependent diffusiophoretic mobility of polystyrene colloids in NaCl gradients, confirming that diffusiophoretic migration velocities are proportional to gradients in the logarithm of electrolyte concentration. We then perform the first direct measurement of the concentration-dependent "solvophoretic" mobility of colloids in ethanol-water gradients, whose dependence on concentration and gradient strength was not known either theoretically or experimentally, but which our measurements reveal to be proportional to the gradient in the logarithm of ethanol mole fraction. Finally, we examine solvophoretic migration under a variety of qualitatively distinct chemical gradients, including solvents that are miscible or have finite solubility with water, an electrolyte for which diffusiophoresis proceeds down concentration gradients (unlike for most electrolytes), and a nonelectrolyte (sugar). Our technique enables the direct characterization of diffusiophoretic mobilities of various colloids under various solvent and solute gradients, analogous to the electrophoretic ζ-potential measurements that are routinely used to characterize suspensions. We anticipate that such measurements will provide the feedback required to test and develop theories for solvophoretic and diffusiophoretic migration and ultimately to the conceptual design and engineering of particles that respond in a desired way to their chemical environments

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Induced charge electroosmosis micropumps using arrays of Janus micropillars.

We report on a microfluidic AC-driven electrokinetic pump that uses Induced Charge Electro-Osmosis (ICEO) to generate on-chip pressures. ICEO flows occur when a bulk electric field polarizes a metal object to induce double layer formation, then drives electroosmotic flow. A microfabricated array of metal-dielectric Janus micropillars breaks the symmetry of ICEO flow, so that an AC electric field applied across the array drives ICEO flow along the length of the pump. When pumping against an external load, a pressure gradient forms along the pump length. The design was analyzed theoretically with the reciprocal theorem. The analysis reveals a maximum pressure and flow rate that depend on the ICEO slip velocity and micropillar geometry. We then fabricate and test the pump, validating our design concept by demonstrating non-local pressure driven flow using local ICEO slip flows. We varied the voltage, frequency, and electrolyte composition, measuring pump pressures of 15-150 Pa. We use the pump to drive flows through a high-resistance microfluidic channel. We conclude by discussing optimization routes suggested by our theoretical analysis to enhance the pump pressure

Induced charge electroosmosis micropumps using arrays of Janus micropillars.

We report on a microfluidic AC-driven electrokinetic pump that uses Induced Charge Electro-Osmosis (ICEO) to generate on-chip pressures. ICEO flows occur when a bulk electric field polarizes a metal object to induce double layer formation, then drives electroosmotic flow. A microfabricated array of metal-dielectric Janus micropillars breaks the symmetry of ICEO flow, so that an AC electric field applied across the array drives ICEO flow along the length of the pump. When pumping against an external load, a pressure gradient forms along the pump length. The design was analyzed theoretically with the reciprocal theorem. The analysis reveals a maximum pressure and flow rate that depend on the ICEO slip velocity and micropillar geometry. We then fabricate and test the pump, validating our design concept by demonstrating non-local pressure driven flow using local ICEO slip flows. We varied the voltage, frequency, and electrolyte composition, measuring pump pressures of 15-150 Pa. We use the pump to drive flows through a high-resistance microfluidic channel. We conclude by discussing optimization routes suggested by our theoretical analysis to enhance the pump pressure

Microfluidic Microdialysis: Spatiotemporal Control over Solution Microenvironments Using Integrated Hydrogel Membrane Microwindows

We present a powerful and versatile technique that enables exquisite spatial and temporal control over local solution chemistry in microfluidic devices. Using a microscope and a UV lamp, we use projection lithography to photopolymerize thin (10–25  μm) hydrogel membrane “microwindows” (HMMs) into standard microfluidic devices. These microwindows are permeable to solute and solvent diffusion and to electric fields, yet act as rigid walls from the standpoint of fluid flow. Reservoirs of solution may thus be rapidly imposed, switched, and maintained on one side of a HMM using standard microfluidic techniques, provoking changes in solution conditions on the other side without active mixing, stirring, or diluting. We highlight three paradigmatic experimental capabilities enabled by HMMs: (1) rapid dialysis and swapping of solute and/or solvent, (2) stable and convection-free localized concentration gradients, and (3) local electric permeability. The functional versatility of hydrogel microwindow membranes, coupled with the ease and speed of their fabrication and integration into simple microchannels or multilayer devices, will open a variety of novel applications and studies in a broad range of fields

Direct Measurements of Colloidal Solvophoresis under Imposed Solvent and Solute Gradients.

We describe a microfluidic system that enables direct visualization and measurement of diffusiophoretic migration of colloids in response to imposed solution gradients. Such measurements have proven difficult or impossible in macroscopic systems due to difficulties in establishing solution gradients that are sufficiently strong yet hydrodynamically stable. We validate the system with measurements of the concentration-dependent diffusiophoretic mobility of polystyrene colloids in NaCl gradients, confirming that diffusiophoretic migration velocities are proportional to gradients in the logarithm of electrolyte concentration. We then perform the first direct measurement of the concentration-dependent "solvophoretic" mobility of colloids in ethanol-water gradients, whose dependence on concentration and gradient strength was not known either theoretically or experimentally, but which our measurements reveal to be proportional to the gradient in the logarithm of ethanol mole fraction. Finally, we examine solvophoretic migration under a variety of qualitatively distinct chemical gradients, including solvents that are miscible or have finite solubility with water, an electrolyte for which diffusiophoresis proceeds down concentration gradients (unlike for most electrolytes), and a nonelectrolyte (sugar). Our technique enables the direct characterization of diffusiophoretic mobilities of various colloids under various solvent and solute gradients, analogous to the electrophoretic ζ-potential measurements that are routinely used to characterize suspensions. We anticipate that such measurements will provide the feedback required to test and develop theories for solvophoretic and diffusiophoretic migration and ultimately to the conceptual design and engineering of particles that respond in a desired way to their chemical environments

Induced charge electroosmosis micropumps using arrays of Janus micropillars

We report on a microfluidic AC-driven electrokinetic pump that uses Induced Charge Electro-Osmosis (ICEO) to generate on-chip pressures. ICEO flows occur when a bulk electric field polarizes a metal object to induce double layer formation, then drives electroosmotic flow. A microfabricated array of metal-dielectric Janus micropillars breaks the symmetry of ICEO flow, so that an AC electric field applied across the array drives ICEO flow along the length of the pump. When pumping against an external load, a pressure gradient forms along the pump length. The design was analyzed theoretically with the reciprocal theorem. The analysis reveals a maximum pressure and flow rate that depend on the ICEO slip velocity and micropillar geometry. We then fabricate and test the pump, validating our design concept by demonstrating non-local pressure driven flow using local ICEO slip flows. We varied the voltage, frequency, and electrolyte composition, measuring pump pressures of 15-150 Pa. We use the pump to drive flows through a high-resistance microfluidic channel. We conclude by discussing optimization routes suggested by our theoretical analysis to enhance the pump pressure