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.

Microfluidic devices are electrical/mechanical systems that offer the ability to work with minimal sample volumes, short reactions times, and have the possibility to perform massive parallel operations. An important application of microfluidics is blood rheology in microdevices, which has played a key role in recent developments of lab-on-chip devices for blood sampling and analysis. The most popular and traditional method to fabricate these types of devices is the polydimethylsiloxane (PDMS) soft lithography technique, which requires molds, usually produced by photolithography. Although the research results are extremely encouraging, the high costs and time involved in the production of molds by photolithography is currently slowing down the development cycle of these types of devices. Here we present a simple, rapid, and low-cost nonlithographic technique to create microfluidic systems for biomedical applications. The results demonstrate the ability of the proposed method to perform cell free layer (CFL) measurements and the formation of microbubbles in continuous blood flow.

Several studies have already demonstrated that it is possible to perform blood flow studies in microfluidic systems fabricated by using low-cost techniques. However, most of these techniques do not produce microchannels smaller than 100 microns and as a result they have several limitations related to blood cell separation. Recently, manufacturers have been able to produce milling tools smaller than 100 microns, which consequently have promoted the ability of micromilling machines to fabricate microfluidic devices able to perform separation of red blood cells (RBCs) from plasma. In this work, we show the ability of a micromilling machine to manufacture microchannels with dimensions down to 30 microns. Additionally, we show for the first time the ability of the proposed microfluidic device to enhance the cell-free layer close to the walls, leading to perform partial separation of RBCs from plasma.

Several experimental techniques were performed in the past years using in vitro environments, in an attempt to not only understand the blood flow behaviour in microcirculation but also develop microfluidic devices as an alternative clinical methodology to detect blood diseases. Hence, the visualization and measurement of red blood cells (RBCs) flowing in a microfluidic device are important to provide not only essential information about hydrodynamic characteristics of the blood but also vital information to diagnose the initial symptoms of diseases during clinical investigations. For instance, RBC rigidity has been correlated with myocardial infarction, diabetes mellitus, hypertension, and also other haematological disorders and diseases that affect RBC deformation more directly, such as, hereditary spherocytosis, sickle cell anaemia and malaria. Regarding a better understanding of the RBCs deformation and motion, we present in this paper a compilation of studies made in our research group, using several microfluidic devices with different microchannel geometries and fabrication techniques (i.e., soft-lithography, xurography and hybrids) that focus in the shear and extensional flow behaviour, either in healthy or chemically stiffed RBCs.

Red blood cells (RBCs) deformability is a high relevant mechanical property, whose variations are associated with some diseases, such as diabetes and malaria. Therefore, the present study aims to compare different image processing methods for assessing the RBCs deformability in a continuous flow, measured in a polydimethylsiloxane (PDMS) microchannel composed by 15 μm spacing inner pillars. The images were acquired with a high speed camera and analyzed with ImageJ software for tracking and measuring the RBCs deformation index (DI). Additionally, to understand the performance of the software, it was performed a comparison between different image processing tools provided by ImageJ and the best methods for the deformation measurements were selected, considering the measured RBCs number and their DIs. The results show that those image methods significantly affect the number of measured RBCs and their DIs and, therefore, the studies focused on the deformability measurements need to take into account the effect of the image processing methods for avoiding loss of relevant information in the images.

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.

Axial migration of individual red blood cells (RBCs) is a well known effect that happens in microchannels with dimensions below 300 μm. This effect is known as the Fåhræus-Lindqvist effect gives place to a cell free layer (CFL) of plasma located around the walls of the microchannel [1]. Several flow studies regarding the CFL measurement exist in in vitro [1-3] environments. However, the manipulation of in vitro blood is often difficult due to the ethical, economical and safety issues involved. As a result, the development of blood analogue solutions is needed to improve our understanding regarding the blood flow behaviour in microchannels and microvessels. Although several studies using one-phase Newtonian and non-Newtonian blood analogues can be found in the literature [4, 5], there are few works related with the use of particulate solutions in which the particles mimic the RBCs behaviour. The present work proposes a particulate-viscoelastic solution made of xanthan gum and dextran with rigid polymethylmethacrylate (PMMA) spherical particles able to mimic the effect of CFL formation that frequently happens in in vitro blood flow systems. Flow visualizations through hyperbolic contraction microchannels were used to observe the CFL originated by the proposed blood analogue fluid. Additionally, this CFL was compared with the plasma layer formed by an in vitro blood sample.

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.

Intracranial aneurysm is a local dilatation of an intracranial artery with high risk of rupture and death. Although it is generally accepted that the weakening of the arterial wall is the main cause for the rupture of an aneurysm, it still no consensus about the reasons for its creation, expansion and rupture. In particular, what is the role played by the blood flow in these phenomena. In this way, the aim of this work is the in vitro mechanical assessment of the wall expansion, namely the displacements, deformations and strains occurring in a saccular intracranial aneurysm model, when subjected to different flow rates. To obtain new insights into the mechanisms involved in the aneurysm rupture, a 3D-VicTM Digital Image Correlation System was used and validated with a finite element analysis. The wall expansion results have revealed that the displacements, deformations and principal strains are directly related to the internal pressure caused by the fluid on the wall of the aneurism. These findings were especially observed in the weakened areas of the aneurysm model, where the wall was thinner. Furthermore, the technique used in this study has shown to be a potential method to validate numerical simulations of aneurysms, allowing the future performance of more complex and realistic haemodynamic studies.

Polydimethylsiloxane (PDMS), due to its remarkable properties, is a suitable polymer for the production of microparticles with industrial and medical applications. The micro-sized PDMS liquid droplets suffer a pronounced shrinkage while curing to turn into solid particles. In this article, we report the calibration of the shrinkage phenomenon in the production of PDMS microparticles. Our results show that this shrinkage does not depend on the amount of curing agent in the PDMS precursor or on the addition of micro/nanoparticles to the mixture, but on the surface effects due to the relatively large droplet surface-to-volume ratio. Moreover, we have also investigated the addition of colour to the particles to improve their visualization/detection. The addition of colour by using pigments enhances the visualization of the contours of the PDMS microparticles, and reveals the capability of this technique to microencapsulate micro/nanoparticles in PDMS spheres with diameters below 10 μm. We demonstrate that the technique used in this work is able to work with a wide range of viscosities obtaining an acceptable degree of monodispersity.