Microfluidic system for cell separation and deformation assessment based on biomimetic and acoustofluidic methods

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Publications

A simple microfluidic device for the deformability assessment of blood cells in a continuous flow

Publication . Rodrigues, Raquel Oliveira; Pinho, Diana; Faustino, Vera; Lima, Rui A.

Blood flow presents several interesting phenomena in microcirculation that can be used to develop microfluidic devices capable to promote blood cells separation and analysis in continuous flow. In the last decade there have been numerous microfluidic studies focused on the deformation of red blood cells (RBCs) flowing through geometries mimicking microvessels. In contrast, studies focusing on the deformation of white blood cells (WBCs) are scarce despite this phenomenon often happens in the microcirculation. In this work, we present a novel integrative microfluidic device able to perform continuous separation of a desired amount of blood cells, without clogging or jamming, and at the same time, capable to assess the deformation index (DI) of both WBCs and RBCs. To determine the DI of both WBCs and RBCs, a hyperbolic converging microchannel was used, as well as a suitable image analysis technique to measure the DIs of these blood cells along the regions of interest. The results show that the WBCs have a much lower deformability than RBCs when subjected to the same in vitro flow conditions, which is directly related to their cytoskeleton and nucleus contents. The proposed strategy can be easily transformed into a simple and inexpensive diagnostic microfluidic system to simultaneously separate and assess blood cells deformability.

2015Journal article

Open access

Biomedical microfluidic devices by using low-cost fabrication techniques: a review

Publication . Faustino, Vera; Catarino, Susana; Lima, Rui A.; Minas, Graça

One of the most popular methods to fabricate biomedical microfluidic devices is by using a soft-lithography technique. However, the fabrication of the moulds to produce microfluidic devices, such as SU-8 moulds, usually requires a cleanroom environment that can be quite costly. Therefore, many efforts have been made to develop low-cost alternatives for the fabrication of microstructures, avoiding the use of cleanroom facilities. Recently, low-cost techniques without cleanroom facilities that feature aspect ratios more than 20, for fabricating those SU-8 moulds have been gaining popularity among biomedical research community. In those techniques, Ultraviolet (UV) exposure equipment, commonly used in the Printed Circuit Board (PCB) industry, replaces the more expensive and less available Mask Aligner that has been used in the last 15 years for SU-8 patterning. Alternatively, non-lithographic low-cost techniques, due to their ability for large-scale production, have increased the interest of the industrial and research community to develop simple, rapid and low-cost microfluidic structures. These alternative techniques include Print and Peel methods (PAP), laserjet, solid ink, cutting plotters or micromilling, that use equipment available in almost all laboratories and offices. An example is the xurography technique that uses a cutting plotter machine and adhesive vinyl films to generate the master moulds to fabricate microfluidic channels. In this review, we present a selection of the most recent lithographic and non-lithographic low-cost techniques to fabricate microfluidic structures, focused on the features and limitations of each technique. Only microfabrication methods that do not require the use of cleanrooms are considered. Additionally, potential applications of these microfluidic devices in biomedical engineering are presented with some illustrative examples.

2016Journal article

Open access

Red blood cells radial dispersion in blood flowing through microchannels: the role of temperature

Publication . Pinho, Diana; Rodrigues, Raquel Oliveira; Faustino, Vera; Yaginuma, Tomoko; Exposto, José; Lima, Rui A.

The behavior of suspensions of individual blood cells, such as red blood cells (RBCs), flowing through microvessels and microfluidic systems depend strongly on the hematocrit (Hct), microvessel topology and cell properties. Although it is well known that blood rheological properties are temperature dependent, to the best of our knowledge no work has studied the role of the temperature on the RBCs dispersion. A powerful way to investigate this latter effect is through a high-speed video microscopy system, which provides detailed flow measurements of each individual RBC. Hence, the effect of temperature on the RBCs dispersion flowing through a 100 μm glass capillary was examined by means of a confocal micro-PTV system. Hundreds of labeled RBCs were tracked at moderate Hct (12%) and at four different temperatures, i.e., 25 °C, 32 °C, 37 °C and 42 °C. The results yielded an enhancement of the RBCs diffusion as the temperature increases. Hence, our findings show that RBCs radial dispersion is temperature dependent and as a result the temperature should not be ignored in future blood flow studies. We believe that this finding is important for a better understanding of blood mass transport mechanisms under both physiological and pathological conditions.

2016Journal article

Restricted access

A passive microfluidic device based on crossflow filtration for cell separation measurements: a spectrophotometric characterization

Publication . Faustino, Vera; Catarino, Susana; Pinho, Diana; Lima, Rui A.; Minas, Graça

Microﬂuidic devices have been widely used as a valuable research tool for diagnostic applications. Particularly, they have been related to the successful detection of different diseases and conditions by assessing the mechanical properties of red blood cells (RBCs). Detecting deformability changes in the cells and being able to separate those cells may be a key factor in assuring the success of detection of some blood diseases with diagnostic devices. To detect and separate the chemically modiﬁed RBCs (mimicking disease-infected RBCs) from healthy RBCs, the present work proposes a microﬂuidic device comprising a sequence of pillars with different gaps and nine different outlets used to evaluate the efﬁciency of the device by measuring the optical absorption of the collected samples. This latter measurement technique was tested to distinguish between healthy RBCs and RBCs chemically modiﬁed with glutaraldehyde. The present study indicates that it was possible to detect a slight differences between the samples using an optical absorption spectrophotometric setup. Hence, the proposed microﬂuidic device has the potential to perform in one single step a partial passive separation of RBCs based on their deformability.

2018Journal article

Open access

Label-free multi-step microfluidic device for mechanical characterization of blood cells: diabetes type II

Publication . Pinho, Diana; Faustino, Vera; Catarino, Susana; Pereira, Ana I.; Minas, Graça; Pinho, Fernando T.; Lima, Rui A.

The increasing interest to establish significant correlations between blood cell mechanical measurements and blood diseases, has led to the promotion of microfluidic devices as attractive clinical tools for potential use in diagnosis. A multi-step microfluidic device able to separate red and white blood cells (RBCs and WBCs) from plasma and simultaneously measure blood cells deformability (5 and 20% of hematocrit) is presented in this paper. The device employs passive separation based on the cross-flow filtration principle, introduced at each daughter channel. At the outlets, hyperbolic geometries allow single-cell deformability analysis. The device was tested with blood from five healthy and fifteen diabetic type II voluntary donors. The results have shown that WBCs have lower deformability than RBCs, and no significant differences were observed in WBCs from healthy and pathological blood samples. In contrast, RBCs have shown significant differences, with pathological cells exhibiting lower deformability. Shear rheology has shown that blood from patients with type II diabetes has higher viscosity than blood from healthy donors. This microfluidic device has demonstrated the ability to reduce cell concentration at the outlets down to 1%, an ideal cell concentration for assessing the blood cells deformability, under healthy and pathological conditions. The results provide new insights and quantitative information about the hemodynamics of in vitro type II diabetes mellitus RBCs. Thus, such device can be a promising complement in clinical diagnosis and biological research as part of an integrated blood-on-a-chip system.

2022Journal article

Open access