Abstract

Parallel High-Throughput Single-Cell Printing Platform for Optoporation-Mediated Large Cargo Delivery

Gayathri R, PhD Scholar (Student), Indian Institute of Technology, Madras

Effective intracellular delivery of biomolecules is crucial in analysing and engineering cell functions, for applications in therapeutic development, diagnostics, and drug delivery towards personalised medicine. Traditional bulk cell culture models employed so far for intracellular delivery, overlook cell population heterogeneity, while single-cell patterning offers robust statistical analyses without compromising cellular variability. In this work, a high-throughput single-cell patterning approach using microcontact printing is employed, where a micro-pillar stamp is used to imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters. This approach is universal for any protein-cell combination and has 97-99 % patterning efficiency. This single-cell patterning was used as a platform for massively parallel optoporation-mediated intracellular delivery of small to very large biomolecules such as PI dye (668 Da), Dextran (3000 MW), SiRNA (20-24 bp), and enzyme (464 kDa). An easy-to-fabricate and simple-to-use 2D array of titanium micro-dish device is used to facilitate near infrared mediated optoporation. which disrupts the cell plasma membrane, allowing biomolecules to enter into cells with high efficiency (~96±2%) and cell viability (~98±1%). Our compact, robust, and simple-to-print approach has potential for single-cell therapy and diagnostics research, providing high specificity and subcellular accuracy in single-cell level biological processes.