An easy-to-handle microfluidic device suitable for immunohistochemical procedures in mammalian cells grown under flow conditions

Abstract

Microfluidics, the technology that manipulates small amount of fluids in microscale complex devices, has undergone a remarkable development during the last decade, by targeting a significant range of applications, including biological tests and single-cell analysis, and by displaying many advantages such as reduced reagent consumption, decreased costs and faster analysis. Furthermore, the introduction of microfluidic tools has revolutionized the study of vascular functions, because the controlled three-dimensional environment and the continuous perfusion provided by the microdevice allow simulating the physiological characteristics of the circulatory system. Researchers interested in the study of vascular physiology, however, are often hampered by the difficulty in handling reduced number of cells after growth in these devices. This work shows how to apply different protocols commonly used in biology, such as the immunofluorescence technique, to cells grown in reversibly-bound microfluidic devices, obtaining results comparable to those retrieved under static conditions in multiwells. In this way, we are able to combine the advantages of microfluidic, i.e., application of continuous flow and shear stress, with classical protocols for the study of endothelial cells.

Influence of shear stress and size on viability of endothelial cells exposed to gold nanoparticles

“…This table is provided to facilitate comparison with literature data, since the concentrations of colloidal solutions were often reported with only one of the proposed scales. Table 1 also reported the same data for the second batch of Au NP (batch 13 nm) previously investigated by us and characterized by smaller dimensions (average diameter 13 nm ± 3 nm) (Fede et al 2015 ).…”

“…Table 1 also reported the same data for the second batch of Au NP (batch 13 nm) previously investigated by us and characterized by smaller dimensions (average diameter 13 nm ± 3 nm) (Fede et al 2015 ).…”

“…This distribution, together with the weighted amount of gold used to synthesize the NP, are the data used to quantify NP concentration in solution. In particular, as previously described by us, the whole distribution of sizes was used to determine the number of NP per milliliter as well as the NP molar concentration (Fede et al 2015 ). A summary of the concentrations of Au NP employed in this work expressed with different units of measurements is given in Table 1 .…”

“…Recently, we compared the change in viability of HUVEC exposed for 24 h with varying doses of citrate-capped Au NP (13 nm diameter) under flow vs. static conditions (Fede et al 2015 ). HUVEC were grown inside multiwells (static exposure) or inside a linear MFD (flow exposure).…”

A versatile lab-on-a-chip tool for modeling biological barriers

“…Previously, the rat cell line RBE4 was studied for a similar purpose [17,36]. Biochips modeling the vascular system also widely use peripheral endothelial cells [37][38][39][40].…”