Dynamic Measurement of Nanoflows: Realization of an Optofluidic Flow Meter to the Nanoliter-per-Minute Scale.

The ultimate performance of flow-based measurements in microfluidic systems is currently limited by their accuracy at the nanoliter-per-minute scale. Improving such measurements (especially in contexts that require continuous monitoring) is challenging because of constraints associated with shrinking system geometries and limitations imposed by making precise measurements of smaller quantities in real time. A particularly interesting limit is the relative uncertainty as flow approaches zero, which diverges for most measurement methods. To address these problems, we have developed an optofluidic measurement system that can deliver and record light in a precise interrogation region of a microfluidic channel. The system utilizes photobleaching of fluorophore dyes in the bulk flow and can identify zero flow to better than 1 nL/min absolute accuracy. The technique also provides an independent method for determining nonzero flow rates based on a robust scaling relationship between the fluorescence emission and flow. Together, these two independent approaches enable precise measurement of flow to within 5% accuracy down to 10 nL/min and validation of flow control to within 5% uncertainty down to 2 nL/min. We also demonstrate that our technique can be used to extend a calibrated flow meter well below its specified range (e.g., 500 nL/min) and to make dynamic measurements of similar relative uncertainties to the calibrated meter, which would have otherwise expanded significantly in this regime.

Flow fluctuations in wormlike micelle fluids.

We investigate the unstable flow of wormlike micelle solutions in pressure driven capillary flow, with a focus on the effect of entrance geometry on the fluid fluctuations. The flow is measured at different points in the capillary using particle image velocimetry while simultaneously measuring the pressure drop across the entire capillary. The fluctuations are characterized by rapid flow rate jumps that correspond with a decrease in the pressure drop followed by a longer recovery period. Velocimetry measurements in the entrance region show a transition to unstable flow above a critical flow rate, where large flow circulations are observed in the tapered geometry and localized jets are observed in an abrupt contraction. The transition to this unstable flow is shown to occur at a similar dimensionless extension rate normalized by the micelle relaxation time. A rapid breakdown in micelle alignment is observed in polarized light microscopy at the onset of the flow rate jump, indicating the importance of rapid micelle structural changes on the fluctuations. We characterize the system by analyzing the power spectral densities and develop a dynamical systems model to describe the relationship between pressure and flow rate. These developments provide understanding to control flow fluctuations and motivation for more detailed study of the coupling of fluid microstructure transitions and flow fluctuations.

Essential role for proteinase-activated receptor-2 in arthritis.

Using physiological, pharmacological, and gene disruption approaches, we demonstrate that proteinase-activated receptor-2 (PAR-2) plays a pivotal role in mediating chronic inflammation. Using an adjuvant monoarthritis model of chronic inflammation, joint swelling was substantially inhibited in PAR-2-deficient mice, being reduced by more than fourfold compared with wild-type mice, with virtually no histological evidence of joint damage. Mice heterozygous for PAR-2 gene disruption showed an intermediate phenotype. PAR-2 expression, normally limited to endothelial cells in small arterioles, was substantially upregulated 2 weeks after induction of inflammation, both in synovium and in other periarticular tissues. PAR-2 agonists showed potent proinflammatory effects as intra-articular injection of ASKH95, a novel synthetic PAR-2 agonist, induced prolonged joint swelling and synovial hyperemia. Given the absence of the chronic inflammatory response in the PAR-2-deficient mice, our findings demonstrate a key role for PAR-2 in mediating chronic inflammation, thereby identifying a novel and important therapeutic target for the management of chronic inflammatory diseases such as rheumatoid arthritis.

Impaired B and T cell antigen receptor signaling in p110delta PI 3-kinase mutant mice.

Class IA phosphoinositide 3-kinases (PI3Ks) are a family of p85/p110 heterodimeric lipid kinases that generate second messenger signals downstream of tyrosine kinases, thereby controlling cell metabolism, growth, proliferation, differentiation, motility, and survival. Mammals express three class IA catalytic subunits: p110alpha, p110beta, and p110delta. It is unclear to what extent these p110 isoforms have overlapping or distinct biological roles. Mice expressing a catalytically inactive form of p110delta (p110delta(D910A)) were generated by gene targeting. Antigen receptor signaling in B and T cells was impaired and immune responses in vivo were attenuated in p110delta mutant mice. They also developed inflammatory bowel disease. These results reveal a selective role for p110delta in immunity.

Effect of protease-activated receptor-2 deficiency on allergic dermatitis in the mouse ear.

To investigate the involvement of protease-activated receptor-2 (PAR-2) in allergic dermatitis, we generated PAR-2-deficient (PAR-2(-/-)) mice. Ear thickness, contact hypersensitivity (CH) induced by topical application of picryl chloride (PC) or oxazolone (Ox) after sensitization, and vascular permeability after ear passive cutaneous anaphylaxis (PCA) were compared between wild-type (WT) and PAR-2(-/-) mice. Ear thickness was almost the same in untreated WT and PAR-2(-/-) mice. Topical application of PC or Ox thickened the ears at 6, 24 and 48 h after challenge with a peak at 24 h in WT mice. In PAR-2(-/-) mice, the ear swelling induced by both PC and Ox was suppressed at every time point, and significant inhibition was found at 24 h in PC-induced CH and at 24 and 48 h in Ox-induced CH. Histopathological observation of the ears at 24 h after challenge revealed that PC- or Ox-induced ear edema and infiltration of inflammatory cells in WT mice were greatly attenuated in PAR-2(-/-) mice. The vascular permeability in the ears after PCA was not different between WT and PAR-2(-/-) mice. These results strongly suggest that PAR-2 plays a crucial role in type IV allergic dermatitis but not in type I allergic dermatitis.