Precision Multi-site Cellular Dosing using a Membrane-Based Laminar-Flow Device

DESCRIPTION (provided by applicant): The proposed Phase I SBIR project is to show the feasibility of developing an affordable commercial instrument that provides laminar-flow fluidic interfaces to cell cultures at multiple coordinates in real time. The instrument will satisfy a growing, unmet need for a method that allows superior spatio-temporal control over the interactions between chemical dosants and cells using inexpensive materials and equipment. Spatio-temporal control is critical to examining interactions between chemical dosants and cells, especially in studies focused on polarized responses within individual cells such as directional chemotaxis. Minotaur Technologies' approach is based on the rapid laser-mediated creation of apertures within a membrane that serves as a support for cell growth and also separates two flow chambers - one of which is a cell medium chamber, and the other, a dosing chamber. This use of laser-induced dielectric breakdown as a means to machine surface pores allows for a precisely-directed laminar flow stream from the higher pressure dosing chamber into the cell environment such that only certain selected cells or cell regions are dosed with the desired chemical species. In the Phase I project Minotaur will demonstrate the feasibility of dosing cells with a dosant region resolution down to 2 micrometers and show the benefits of the technology for important neuroscience applications. Although we understand much of what goes on inside cells at a molecular level, understanding of how cells function as a whole lags far behind. Frequently, the study of living cells is hampered by lack of appropriate technologies. Scientific advances depend not only on new ideas, but also on technological advances that make conceptual shifts possible. Chemical dosing of live cells with agents that selectively alter cellular function in a controlled fashion provides a vital means to study cellular differentiation, growth, and death. The instrument developed will be applicable to a broad range of cell biology problems in neuroscience and beyond, including chemotaxis, stem-cell differentiation and in vitro testing of cellular response to emerging treatments. Public Health Relevance: A fundamental component of cellular organisms is the ability of cells to sense and respond to their environment. Elucidating the mechanisms of cellular responses to external stimuli and how cell-to-cell signaling pathways coordinate cellular activities is key to understanding how cells work. The improved understanding of cellular responses in both healthy and diseased states that may be gained from use of the proposed instrument could lead to development of more effective medicines and methods of diagnosis.