Sphyga: A multiparameter open source tool for fabricating smart and tunable hydrogel microbeads

Hydrogel microbeads are used in many biological applications, particularly for cell, protein or drug encapsulation. Although there are several methods for fabricating microbeads with controlled shapes and dimensions, many are limited to a small range of materials or sizes. We describe a compact open source tool - the spherical hydrogel generator (Sphyga) - for the fabrication of highly reproducible hydrogel based microbeads with predictable shapes and diameters ranging from 100 to 2000 μm. The unique feature of the system is the ability to modulate multiple parameters independently, so as to create a wide range of working conditions for fabricating tailored microbeads. Hence, by combining the different fabrication parameters, hydrogel beads with chosen shapes, sizes and materials can be generated with Sphyga. A multiparameter working-window was obtained by fixing the concentration of the base material, alginate, and varying the viscosity of the solution along with Sphyga's fabrication parameters (needle size, external air pressure, and material outflow). To validate the multiparameter working window, components such as proteins, cells, dyes and nanoparticles were also used to fabricate composite microbeads. The results show that the architecture of hydrogel microbeads can be engineered by considering the viscosity of the initial solution, which depends principally on the pH and composition of alginate solution. Coupled with Sphyga's multiple working parameters, material viscosity can then be used to tune hydrogel domains and thereby generate complex biologically relevant microenvironments for many biomedical applications.