Abstract
The ability to deliver foreign molecules into a single living cell with high transfection efficiency and high cell viability is of great interest in cell biology for applications in therapeutic development, diagnostics, and drug delivery towards personalized medicine. Various physical delivery methods have long demonstrated the ability to deliver cargo molecules directly to the cytoplasm or nucleus and the mechanisms underlying most of the approaches have been extensively investigated. However, most of these techniques are bulk approaches that are cell-specific and have low throughput delivery. In comparison to bulk measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. To elucidate distinct responses during cell genetic modification, methods to achieve transfection at the single-cell level are of great interest. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. This review article aims to cover various microfluidic-based physical methods for single-cell intracellular delivery such as electroporation, mechanoporation, microinjection, sonoporation, optoporation, magnetoporation, and thermoporation and their analysis. The mechanisms of various physical methods, their applications, limitations, and prospects are also elaborated.
Highlights
The cell is the most fundamental independent unit of life which transmits information through molecules
The results show that DNA and messenger RNA (mRNA) are significantly dependent on the application of the electric field, while protein delivery is more dependent on mechanical membrane disruption methods
Sonoporationisisa aphysical physical approach of membrane disruption, in which an ultrais applied in the presence of microbubbles that acts as cavitation nuclei, which forces the sound is applied in the presence of microbubbles that acts as cavitation nuclei, which cell membrane resulting in disruption and the creation transient pores to forces the cell membrane resulting in disruption and theofcreation of membrane transient membrane allow biomolecules into theinto cell the includes ultrasound contrast pores to allow biomolecules cell The
Summary
The cell is the most fundamental independent unit of life which transmits information through molecules. Researchers have been conducting experiments for decades for developing and adapting molecules into prospective cargo to be delivered into the intracellular environment [4] The majority of these cargos are membrane-impermeable and need physical energy for intracellular delivery [5]. Intracellular delivery approaches are broadly divided into two types: (i) By using biological or viral vectors technique and (ii) by using physical techniques (non-viral chemical vectors) to deliver molecules into cell cytoplasm or nucleus. Various microfluidic-based physical methods for single-cell intracellular delivery have been gaining importance and major advancements have been happening in this area of transfection. We critically assess each delivery method comparing their performance parameters such as (i) cell viability and transfection efficiency with different cell types and cargo molecules, (ii) effectiveness of the single-cell approach, (iii) possibility to integrate with current workflows, and (iv) adaptability to changeover between delivery of different cell types, sizes, and morphologies
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