Delivery of Biomolecules into Individual Cells and Subcellular Compartments by Localized Electroporation via Nanopipette

Introducing exogenous biomolecules into individual cells
with precise
control over space, time, and dosage is crucial for both fundamental
and applied biological research. Glass nanopipettes have long been
employed to deliver biomolecules into individual cells; yet, their
reliance on the electrical charge of the target molecule and the need
for penetrating the cellular membrane pose significant limitations.
We demonstrate that voltage pulses applied through a glass nanopipette
in proximity to the cell membrane induce localized electroporation
and generate directional flow, enabling controlled delivery of both
charged and neutral biomolecules into subcellular compartments, e.g.,
the nucleus, without the need for penetrating the cellular membrane.
This approach minimizes cell damage and preserves cell viability,
even after multiple rounds of injection. Our findings will serve as
a reference for the design of novel nanopipette methods, contributing
to the newly established field of spatiotemporal analysis of live
cells.