Microfluidic Chemotaxis-Based Cell Immunophenotyping for Ocular Diagnostics

Abstract

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Uveitis and primary intraocular lymphoma (PIOL) are diseases associated with the presence of increased lymphocytes within the eye. With the uveitis and PIOL diseases, there is an invasion of B-lymphocytes and T-lymphocytes, respectively. Each require unique methods for treatment, but the symptoms for each of the diseases are quite similar. Additionally, the physical structures of the two cell types are indistinguishable. The determination of cell type is currently done through an extensive flow cytometry procedure. This requires the exportation of the samples on ice (they change conformation ex vivo after approximately one hour and require extensive care to maintain fidelity), as well as trained personnel to run the instrument and deliver the results. This process is neither time or cost efficient.

Chemotaxis is the directed migration of particular cell types towards gradients of chemicals known as chemoattractants. The phenomenon of chemotaxis can be used for cell separation via competing gradients of T- and B-lymphocyte chemoattractant. Previous work has shown the efficacy of using chemotactic gradients in microfluidic devices. The implementation of this concept on a microfluidic lab-on-a-chip format would ultimately allow for the separation and quantification of the two cell types in a more cost and time effective manner, leading to accurate diagnosis within a matter of minutes.   Our work develops a fabric microfluidic device capable of producing competing gradients of different chemoattractants which will separate cells based on phenotype.  Cell movement is recorded on device and analyzed using cell tracking software.  Analysis of directed cell migration allows for diagnosis.