Inertial separation techniques in a microfluidic system have been widely employed in the field of medical diagnosis for a long time. Despite no requirement of external forces, it requires strong hydrodynamic forces that could potentially cause cell damage or loss during the separation process. This might lead to the wrong interpretation of laboratory results since the change of structures and functional characteristics of cells due to the hydrodynamic forces that occur are not taken into account. Therefore, it is important to investigate the cell viability and damage along with the separation efficacy of the device in the design process. In this study, two inertial separation techniques—spiral microchannel and contraction-expansion array (CEA)—were examined to evaluate cell viability, morphology and intracellular structures using a trypan blue assay (TB), Scanning Electron Microscopy (SEM) and Wright-Giemsa stain. We discovered that cell loss was not significantly found in a feeding system, i.e., syringe, needle and tube, but mostly occurred in the inertial separation devices while the change of cell morphology and intracellular structures were found in the feeding system and inertial separation devices. Furthermore, percentage of cell loss was not significant in both devices (7–10%). However, the change of cell morphology was considerably increased (30%) in spiral microchannel (shear stress dominated) rather than in CEA (12%). In contrast, the disruption of intracellular structures was increased (14%) in CEA (extensional and shear stress dominated equally) rather than spiral microchannel (2%). In these experiments, leukocytes of canine were used as samples because their sizes are varied in a range between 7–12 µm, and they are commonly used as a biomarker in many clinical and medical applications.
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