PepS: An Innovative Microfluidic Device for Bedside Whole Blood Processing before Plasma Proteomics Analyses.

Immunoassays have been used for decades in clinical laboratories to quantify proteins in serum and plasma samples. However, their limitations make them inappropriate in some cases. Recently, mass spectrometry (MS) based proteomics analysis has emerged as a promising alternative method when seeking to assess panels of protein biomarkers with a view to providing protein profiles to monitor health status. Up to now, however, translation of MS-based proteomics to the clinic has been hampered by its complexity and the substantial time and human resources necessary for sample preparation. Plasma matrix is particularly tricky to process as it contains more than 3000 proteins with concentrations spanning an extreme dynamic range (1010). To address this preanalytical challenge, we designed a microfluidic device (PepS) automating and accelerating blood sample preparation for bottom-up MS-based proteomics analysis. The microfluidic cartridge is operated through a dedicated compact instrument providing fully automated fluid processing and thermal control. In less than 2 h, the PepS device allows bedside plasma separation from whole blood, volume metering, depletion of albumin, protein digestion with trypsin, and stabilization of tryptic peptides on solid-phase extraction sorbent. For this first presentation, the performance of the PepS device was assessed using discovery proteomics and targeted proteomics, detecting a panel of three protein biomarkers routinely assayed in clinical laboratories (alanine aminotransferase 1, C-reactive protein, and myoglobin). This innovative microfluidic device and its associated instrumentation should help to streamline and simplify clinical proteomics studies.

Developing a new device for continuously recording, in vivo, the excretion rate of sweat (perspiration) in humans.

BACKGROUND: Some methodologies used for evaluating sweat production and antiperspirants are of a stationary aspect, that is, most often performed under warm (38°C) but resting conditions in a rather short period of time. The aim is to develop an electronic sensor apt at continuously recording sweat excretion, in vivo, during physical exercises, exposure to differently heated environments, or any other stimuli that may provoke sweat excretion. MATERIAL AND METHODS: A sensor (20 cm2 ) is wrapped under a double-layered textile pad. Fixed onto the armpits, these two arrays of electrodes are connected to electronic system through an analog multiplexer. A microcontroller is used to permanently record changes in the conductance between two electrodes during exposure of subjects to different sweat-inducing conditions or to assess the efficacy of applied aluminum hydrochloride (ACH)-based roll-ons at two concentrations (5% and 15%). RESULTS: In vitro calibration, using a NaCl 0.5% solution, allows changes in mV to be related with progressively increased volumes. In vivo, results show that casual physical exercise leads to sweat excretions much higher than in warm environment (37 or 45°C). Only, an exposure to a 50°C environment induced comparable sweat excretion. In this condition, sweat excretions were found similar in both armpits and both genders. Decreased sweat excretions were recorded following applications of ACH, with a dose effect. CONCLUSION: Developing phases of this new approach indicate that usual method or guidelines used to determine sweat excretions in vivo do not reflect true energy expenditure processes. As a consequence, they probably over-estimate the efficacy of antiperspirant agents or formulae.

Separation of Biological Particles in a Modular Platform of Cascaded Deterministic Lateral Displacement Modules.

Deterministic lateral displacement (DLD) has been extensively implemented in the last decade for size-based sample preparation, owing to its high separation performances for a wide range of particle dimensions. However, separating particles from 1 µm to 10 µm in one single DLD device is challenging because of the required diversity of pillar dimensions and inherent fabrication issues. This paper presents an alternative approach to achieve the extraction of E. coli bacteria from blood samples spiked with prostate cancer cells. Our approach consists in cascading individual DLD devices in a single automated platform, using flexible chambers that successively collect and inject the sample between each DLD stage without any external sample manipulation. Operating DLD separations independently enables to maximize the sorting efficiency at each step, without any disturbance from downstream stages. The proposed two-step automated protocol is applied to the separation of three types of components (bacteria, blood particles and cancer cells), with a depletion yield of 100% for cancer cells and 93% for red blood cells. This cascaded approach is presented for the first time with two DLD modules and is upscalable to improve the dynamic range of currently available DLD devices.

Flexible sensors for real-time monitoring of moisture levels in wound dressings.

OBJECTIVE: Few studies have investigated methods to monitor the moisture distribution and filling percentage of dressings during wound management. In this study, a new system allowing moisture monitoring across the wound dressing to be measured is examined. METHOD: The system is composed of a wound bed model with a fluid injection system to mimic exudate flow, a flexible sensor array and data acquisition software. The sensor is composed of 14 flexible electrode arrays, screen-printed on a flexible support and placed on the top of a wound dressing. The system is used to evaluate the performance of a foam-based dressing model. RESULTS: During constant injection of fluid, the wound dressing absorbed moisture at the wound interface throughout the experiment, and expanded as the fluid spread from the wound bed to the edging areas of the foam. The in-time monitoring by the use of the screen-printed electrodes allowed a mapping of the dressing wet surface and estimation of the foam saturation (filling percentage) based on a simple acquisition method without the need to remove the dressing from the wound bed. CONCLUSION: The findings of this study propose a non-invasive method to monitor the filling of the wound dressing and consequently, a potential solution for determining the optimal dressing change during wound management without causing irritation or further damage to the periwound skin.

Purification of complex samples: Implementation of a modular and reconfigurable droplet-based microfluidic platform with cascaded deterministic lateral displacement separation modules.

Particle separation in microfluidic devices is a common problematic for sample preparation in biology. Deterministic lateral displacement (DLD) is efficiently implemented as a size-based fractionation technique to separate two populations of particles around a specific size. However, real biological samples contain components of many different sizes and a single DLD separation step is not sufficient to purify these complex samples. When connecting several DLD modules in series, pressure balancing at the DLD outlets of each step becomes critical to ensure an optimal separation efficiency. A generic microfluidic platform is presented in this paper to optimize pressure balancing, when DLD separation is connected either to another DLD module or to a different microfluidic function. This is made possible by generating droplets at T-junctions connected to the DLD outlets. Droplets act as pressure controllers, which perform at the same time the encapsulation of DLD sorted particles and the balance of output pressures. The optimized pressures to apply on DLD modules and on T-junctions are determined by a general model that ensures the equilibrium of the entire platform. The proposed separation platform is completely modular and reconfigurable since the same predictive model applies to any cascaded DLD modules of the droplet-based cartridge.