Massively Parallel High ‐Throughput Single‐Cell Patterning and Large Biomolecular Delivery in Mammalian Cells Using Light Pulses

High-throughput single-cell patterning is presented for massively parallel optoporation-mediated intracellular delivery of small to very large cargo such as PI dye (668  Da), Dextran (3000 MW), siRNA (20–24 bp), and enzyme (464 kDa), using a titanium micro-dish device. The approach is compact, robust, easy to use and potentially applicable for single-cell therapy and diagnostics. AbstractThe recent advancements of single-cell analysis have significantly enhanced the ability to understand cellular physiology when compared to bulk cellular analysis. Here a massively parallel single-cell patterning and very large biomolecular delivery is reported. Micro-pillar polydimethyl siloxane stamp with different diameters (40 –100 µm with 1 cm × 1 cm patterning area) is fabricated and then imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters. The maximum patterning efficiency is achieved 99.7% for SiHa, 96.75% for L929, and 98.6% for MG63 cell s, for the 100 µm micro-pillar stamp. For intracellular delivery of biomolecules into the patterned cells, a titanium micro-dish device is aligned on top of the cells and exposed by infrared light pulses. The platform successfully delivers small to very large biomolecules such as PI dyes (668 Da) , dextran 3000 Da, siRNA (20-24 bp), and large size enzymes (464 KDa) in SiHa, L929 and MG63 cells. The delivery efficiency for PI dye, Dextran 3000, siRNA, and enzyme...
Source: Small - Category: Nanotechnology Authors: Tags: Research Article Source Type: research