Introduction The modular μSiM is based on a 96-well plate format with a 100 μl open well. While these devices have proven extremely useful for various barrier models including the blood-brain barrier1 and the tendon-vascular interface2, there is a need to transform the open well into a fluidic channel. A closed channel device would enable…
Introduction As discussed in a previous post, we are working to adapt a PDMS flow insert as a manufacturable component. While our priority was maintaining the original flow channel geometry as Mansouri et al. designed, we slightly changed the channel length (5 mm –> 4.6 mm) to accommodate an extra bonding area for the pressure-sensitive…
Introduction A recent collaborative publication from RIT developed a modular µSiM to address the lack of physiological fluid flow in conventional open-well devices (Figure 1). The magnetic latching assembly of this design allows for quick configuration into a fluidic setup. The flow module is a microfabricated PDMS insert that contains a channel over the membrane…
Introduction: Many applications of the µSiM require fluid to be continuously supplied to the microfluidic devices. We use laboratory tubing (typically Cole-Parmer/Tygon Tubing with I.D = 1/16″) that is connected to syringes or peristaltic pumps to do so. However, interfacing the tubing with the current entry and exit ports for the bottom channel is not…
Introduction High throughput imaging techniques in conjunction with in vitro micro-physiological systems (MPS) allow for comprehensive exploration of physiologically relevant phenomena in a living 3D mimetic of a tissue microenvironment. Prior work in this lab has readily demonstrated that vascular tissue mimetics (the µSiM) facilitate real time phase-microscopy visualization of immune cell trafficking in response…
Overview Recently, we performed a series of flow cytometry studies assessing neutrophil (PMN) activation in first generation flow devices. All the relevant data and can be found in a previous post: here. As part of the analysis, we chose to quantify relative PMN activation by comparing isotype control histograms against L-selectin (CD62L) expression histograms. This…
Introduction As part of our continuing evaluation of the flow cell developed by our collaborators at RIT, our next goal is to demonstrate that neutrophils remain inactivated when flown through a first generation device (Figure 1). For this, we studied neutrophil activation as a function of fluid flow induced shear stress based on the physical…
Introduction Static endothelial cell cultures have been routinely performed and optimized on the µSiM platform for multiple membrane and cell types. In order to increase the physiological relevance of our device with tissue vasculature mimetics, we need to incorporate fluid flow to emulate shear stresses experienced by endothelium under physiological circulatory conditions. Recently Mehran has…
Introduction Infection or injury of tissue results in inflammatory signalling that motivates leukocyte recruitment to an inflamed location. Monocytes in particular are implicated in both pro inflammatory signalling through the M1 macrophage phenotype and regulation through the M2 macrophage phenotype. The μSiM devices developed in our lab have yet to assess/optimize monocyte transmigration from the…
Please see the attached .PDF file for a full monocyte isolation protocol. This is for isolating human monocytes from whole blood. Monocyte-Isolation-Protocol