Browsing by Author "Pinto, Elmano"
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- Blood flow in microchannels manufactured by a low cost technique: xurographyPublication . Pinto, Elmano; Taboada, Bruna Rafaela Pereira; Faustino, Vera; Cidre, Diana; Rodrigues, Raquel Oliveira; Miranda, João Mário; Garcia, Valdemar; Dias, Ricardo P.; Lima, Rui A.The xurography is a technique that has been used to make molds to produce microchannels. In contrast to soft lithography [1, 2], xurography uses equipments and materials commonly used in the printing industry, such as cutting plotters, vinyl and other materials. The main advantage of this technique is to fabricate microchannels at a reduced cost [3, 4]. The Fahraeus-Lindqvist effect is a well know phenomenon that happens in microcirculation, where red blood cells (RBCs) have tendency to migrate toward the centre of the microtube resulting in a marginal cell-free layer (CFL) at regions adjacent to the wall [5]. Recently several studies showed strong evidence that the formation of the CFL is affected by the geometry of the microchannel [1, 6, 7] and the physiological conditions of the working fluid, such as the hematocrit (Hct) [2, 8]. The main objective of the present work is to fabricate polydimethysiloxane (PDMS) microchannels by using a soft xurography technique in order perform blood flow studies. Additionally, a high-speed video microscopy system is used to measure the CFL thickness in two different geometries, i. e., bifurcations and confluences.
- Cell-free layer (CFL) measurements in complex geometries: contractions and bifurcationsPublication . Novais, Susana; Pinho, Diana; Bento, David; Pinto, Elmano; Yaginuma, Tomoko; Fernandes, Carla S.; Garcia, Valdemar; Pereira, Ana I.; Lima, José; Mujika, Maite; Oliveira, Mónica S.N.; Dias, Ricardo P.; Arana, Sergio; Lima, Rui A.In this chapter we discuss the cell-free layer (CFL) developed adjacent to the wall of microgeometries containing complex features representative of the microcirculation, such as contractions, expansions, bifurcations and confluences. The microchannels with the different geometries were made of polydimethylsiloxane (PDMS) and we use optical techniques to evaluate the cell-free layer for red blood cells (RBC) suspensions with different hematocrit (Hct). The images are captured using a high-speed video microscopy system and the thickness of the cell free layer was measured using both manual and automatic image analysis techniques. The results show that in in vitro microcirculation, the hematocrit and the geometrical configuration have a major impact on the CFL thickness. In particular, the thickness of the cell-free layer increases as the fluid flows through a contraction-expansion sequence and that this increase is enhanced for lower hematocrit. In contrast, the flow rates tested in this studies did not show a clear influence on the CFL thickness.
- Cell-free layer analysis in a polydimethysiloxane microchannel: A global approachPublication . Pinto, Elmano; Faustino, Vera; Pinho, Diana; Rodrigues, Raquel Oliveira; Lima, Rui A.; Pereira, Ana I.The cell-free layer (CFL) is a hemodynamic phenomenon that has an important contribution to the rheological properti es of blood flowing in microvessels. The present work aims to find the closest function describing RBCs flowing around the cell depleted layer in a polydimethysiloxane (PDMS) microchannel with a diverging and a converging bifurcation. The flow behaviour of the CFL was investigated by using a high-speed video microscopy system where special attention was devoted to its behaviour before the bifurcation and after the confluence of the microchannel. The numerical data was first obtained by using a manual tracking plugin and then analysed using the genetic algorithm approach. The results show that for the majority of the cases the function that more closely resembles the CFL boundary is the sum of trigonometric functions.
- Cell-free layer measurements in a bifurcation microchannel : comparison between a manual and automatic methodsPublication . Bento, David; Pinho, Diana; Pinto, Elmano; Yaginuma, Tomoko; Correia, Teresa Montenegro; Lima, José; Pereira, Ana I.; Lima, Rui A.In the present work, in vitro blood flowing through a bifurcation microchannel was studied. The aim was to measure the Trajectories of the cell-free layer (CFL) by using different methods, i. e., a manual and two automatic methods.
- Cell-free layer measurements in a bifurcation microchannel : comparison between a manual and automatic methodsPublication . Bento, David; Pinho, Diana; Pinto, Elmano; Yaginuma, Tomoko; Correia, Teresa Montenegro; Lima, José; Pereira, Ana I.; Fernandes, Carla S.; Dias, Ricardo P.; Lima, Rui A.In the present work, in vitro blood flowing through a bifurcation microchannel was studied. The aim was to measure the Trajectories of the cell-free layer (CFL) by using different methods, i. e., a manual and two automatic methods.
- Estudo experimental de escoamentos fisiológicos em microcanais fabricados por xurografiaPublication . Pinto, Elmano; Lima, Rui A.; Garcia, Valdemar; Dias, Ricardo P.Este projecto de mestrado em Tecnologia Biomédica é de natureza experimental e possui como principais pontos de interesse a fabricação de microcanais em polidimetilsiloxano (PDMS) a baixo custo e a visualização do escoamento no interior dos microcanais produzidos. Constituem como principais objectivos deste projecto a fabricação de microcanais em PDMS com recurso a tecnologias aplicadas à indústria gráfica e efectuar a sua validação. Propôs-se desenvolver/melhorar uma metodologia de microfabricação de baixo custo, conhecida por xurografia, efectuar a visualização e quantificação de vários fenómenos associados ao escoamento sanguíneo em microcanais. A técnica desenvolvida neste projecto possui como referencia a litografia suave, que por sua vez é a técnica mais utilizada na área da microfabricação. No entanto esta técnica é muito dispendiosa e requer uma sala limpa. Devido à inexistência de uma sala limpa no Instituto Politécnico de Bragança, neste trabalho foi utilizada uma plotter de corte e diversos materiais utilizados na indústria gráfica (vinil, papel adesivo, entre outros) para a produção de um molde e microcanais em PDMS. As geometrias utilizadas para o estudo possuíam larguras com dimensões a variar entre os 150μm e os 1000μm. Estas possuíam ramificações com zonas de bifurcações e consequentes zonas de confluências. Apos a fabricação dos microcanais procedeu-se ao estudo do escoamento de fluidos fisiológicos nesses microcanais. Os fluidos fisiológicos utilizados foram sangue ovino com percentagens de hematócrito entre 1% e 15% em dextrano 40. Foram também testados caudais variáveis entre 5 e 15μL/min. Procedeu-se à visualização do escoamento dos fluidos utilizando um sistema de microscópica e captaram-se várias imagens dos microcanais, nomeadamente antes da bifurcação e depois da confluência. As imagens foram tratadas num software informático, quantificando a espessura da camada livre de células formada junto das paredes e a jusante da confluência. Os resultados obtidos demonstram que a técnica designada por xurografia pode ser utilizada para estudar vários fenómenos fisiológicos existentes na microcirculação. This project of the master's degree in Biomedical Technology is an experimental work and includes as the principal interests the fabrication of microchannels of polydimethylsiloxane (PDMS) at low cost and the ability to study blood flow phenomena within the microchannels produced. The main objective of this project is to fabricate microchannels in PDMS using the technologies applied to the printing industry and validation of their performance. It has been proposed to develop/improve a low cost method of microfabrication, to acquire know how about xurography and to visualize and quantify several physiological phenomena associated with blood flow in microchannels. The technique used as a reference in this project was soft lithography, which is the most used technique in the field of microfabrication. However this technique is very expensive and requires a clean room. Due to the lack of a clean room at the Polytechnic Institute of Bragança, a cutting plotter and various materials used in the printing industry (technique known as xurography) were used to produce the molds and the correspondent microchannels in PDMS. The geometries used in this study had widths with dimensions ranging from 150μm up to 1000μm. These channels had ramifications with bifurcations and confluences regions. After the fabrication of the microchannel the physiological fluid flow in these microchannels were studied. The physiologic fluids used were sheep blood with dextran 40 of which hematocrit percentages were between 1% and 15%. The flow rates varying between 5 and 15μL/min were also tested. The fluid flows were visualized using a microscopic system and captured images of the various microchannels, including especially before the bifurcation and after the confluence. The images were processed with a computer software to quantify the thickness of the cell free layer formed adjacent to the microchannel walls and downstream of the confluence. The results demonstrate that the technique known as xurography can be applied to investigate blood flow phenomena happening in microcirculation.
- Fabricação de microcanais utilizando o método de xurografia: visualização da camada livre de célulasPublication . Pinto, Elmano; Pinho, Diana; Bento, David; Correia, Teresa Montenegro; Garcia, Valdemar; Dias, Ricardo P.; Miranda, João MárioNo presente estudo, pretendeu-se desenvolver/melhorar uma metodologia de microfabricação de baixo custo, conhecida por xurografia e efectuar a visualização e quantificação da camada livre de células para diferentes escoamentos sanguíneos em microcanais.
- Fabricação de microcanais utilizando o método de xurografia: visualização da camada livre de célulasPublication . Pinto, Elmano; Pinho, Diana; Bento, David; Correia, Teresa Montenegro; Garcia, Valdemar; Dias, Ricardo P.; Miranda, João Mário; Lima, Rui A.No presente estudo, pretendeu-se desenvolver/melhorar uma metodologia de microfabricação de baixo custo, conhecida por xurografia e efectuar a visualização e quantificação da camada livre de células (CLC) junto às paredes para diferentes escoamentos sanguíneos em microcanais.
- Flow of red blood cells in microchannel networks: in vitro studiesPublication . Cidre, Diana; Rodrigues, Raquel Oliveira; Faustino, Vera; Pinto, Elmano; Pinho, Diana; Bento, David; Correia, Teresa Montenegro; Fernandes, Carla S.; Dias, Ricardo P.; Lima, Rui A.Blood exhibits unique flow characteristics on micro-scale level, due to the complex biochemical structure of Red Blood Cells (RBCs) and their response to both shear and extensional flow, which influence the rheological properties and flow behavior of blood [1,2]. In the past years, several in vitro studies where made and have revealed some physiologically significant phenomena, such as Fahraeus and Fahraeus-Lindqvist effect, that played a key role in recent developments of lab-onchip devices for blood sampling, analysis and cell culturing. However, the blood flow in microvascular networks phenomena it remains incompletely understood. Thus, it is important to investigate in detail the behavior of RBCs flow occurring at a microchannel network, such as with divergent and convergent bifurcations. Previews in vitro studies in microchannels with a simple divergent and convergent bifurcation, have shown a pronounced cell depleted zone immediately downstream of the apex of the convergent bifurcation. In the present work, by using a highspeed video microscopy system, we investigated the cell depleted zone in a microchannel network. The working fluid used in this study was dextran 40 (Dx40) containing about 10% of haematocrit level (10 Hct) of ovine red blood cells. The high-speed video microscopy system used in our experiments consists of an inverted microscope (IX71, Olympus, Japan) combined with a highspeed camera (i-SPEED LT, Olympus). A syringe pump Apparatus (PHD ULTRATM) with 1 ml syringe (Terumo) was used to push the working fluids through the microchannel network. Additionally, we investigated the effect of the flow rate on the formation of the cell free layer.
- Flow of red blood cells in microchannel networks: in vitro studiesPublication . Cidre, Diana; Rodrigues, Raquel Oliveira; Faustino, Vera; Pinto, Elmano; Pinho, Diana; Bento, David; Fernandes, Carla S.; Dias, Ricardo P.; Lima, Rui A.Human blood is a multiphase biofluid primarily composed by the deformable red blood cells (RBCs) suspended in plasma. Because the complex structure of RBCs, blood exhibits unique flow characteristics on micro-scale level, due to their complex biochemical mechanisms and their response to both shear and extensional flow, which influence the rheological properties and flow behaviour of blood [1,2]. In the past years in vitro blood studies have been extensively performed and some important physiological phenomena, such as Fahraeus and Fahraeus-Lindqvist effect, were revealed [1,3]. This pioneer studies performed by Fahraeus and Fahraeus-Lindqvist in straight glass microchannels [4] revealed that for narrow tubes (diameter<300 μm), the apparent viscosity of blood declines with decreasing diameter. More recently, due to the developments in microscopy, computers and image analysis techniques, several researchers have used new measuring methods to obtain deeper quantitative understanding of the blood flow dynamics, in vitro [5-8] and in vivo experiments [9-10]. The increasing interest by the microfluidic and biomedical communities has also played a key role in several recent developments of lab-on-chip devices for blood sampling, analysis and cell culturing, aimed in a near future, the development of blood diagnostic devices, as an alternative tool to the traditional diagnostic strategies. However, the blood flow in microvascular networks phenomena remains incompletely understood. Thus, it is important to investigate in detail the behaviour of RBCs flow occurring in a microchannel network, such as, with divergent and convergent bifurcations, which mimics the irregular vessel segments linked by numerous diverging and converging bifurcations. Previously, we made in vitro studies in microchannels with a simple divergent and convergent bifurcation, that showed a pronounced cell-free layer (CFL) immediately downstream of the apex of the convergent bifurcation [1,4]. This interesting result led us to the present work, where the CFL in a microchannel network is investigated by using a high-speed video microscopy system in order to further understand the blood flow behaviour in microvessels networks.