Phagocytosis is a biological process in which cell debris, pathogens, or other foreign objects are eliminated. Debris or undesired cells are engulfed by phagocytes and broken down by enzymes for either reuse or excretion. The process of phagocytosis involves many forces which lead to potential deformations of the particulate being engulfed. Measuring the forces involved in phagocytosis would help us to understand the physical mechanisms involved, leading to a better understanding of disease processes. The forces that particulates experience could potentially be studied by employing soft (&lt;10 kPa) microspheres containing discrete fluorophores. Deformation of the spheres could be tracked by measuring the relative displacement of the fluorescent particles in real time during phagocytosis.<br/>We have developed a soft, biocompatible microsphere (2-10 microns in diameter) with discrete fluorescent markers for this purpose. The microspheres are composed of a crosslinked gelatin hydrogel which contains a distribution of small (&lt; 50 nm diameter) fluorescent spheres. To fabricate the microspheres, the fluorescent nanospheres were first added to an aqueous gelatin solution. The resulting solution was mixed vigorously with an organic phase to form a colloidal suspension of small gelatin spheres, which were subsequently crosslinked with glutaraldehyde. An aldehyde functionalization on the surface of the fluorescent nanospheres caused them to crosslink to the gelatin matrix, preventing them from leaching from the final product.<br/>Subsequent work will be separating the microspheres from the excess gelatin and organic phase for eventual use with cell cultures. Other materials may be manipulated using a similar protocol to create a better suited product for the application.

Soft microspheres with discrete fluorescent nanospheres for measuring forces during phagocytosis

253rd National Meeting (2017)

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