Photoporation reimagined: microfluidic paradigm for precision and safe intracellular delivery of large cargos

Abstract

Efficient and safe intracellular delivery remains a major challenge in biotechnology and medicine, especially for large biomolecules such as plasmids, RNP complexes, and proteins. Conventional physical and chemical approaches often compromise cell viability, scalability, or molecular integrity. Photoporation, using laser-induced transient membrane permeabilization, has emerged as a precise, contact-free alternative, enabling localized and minimally invasive delivery. However, traditional photoporation systems are limited by low throughput and poor reproducibility. Integrating photoporation with microfluidic technology addresses these bottlenecks by combining optical precision with hydrodynamic control, enabling uniform exposure, high throughput, and tunable pore dynamics. This review discusses recent advances in microfluidic photoporation platforms, focusing on material-assisted designs, mechanistic insights, and delivery outcomes across diverse cargo sizes. The challenges of laser energy optimization, nanoparticle variability, and clinical scalability are critically examined, alongside emerging opportunities such as AI-assisted feedback control and organ-on-chip validation. By bridging optical engineering, microfluidics, and cell biology, microfluidic photoporation holds transformative potential for next-generation intracellular delivery, cell therapy manufacturing, and precision medicine.