(A) Recipient 143BTK− ρ0 cells were seeded on a 400 μm × 400 μm square to facilitate nanoblade delivery, tracking, and clonal selection.

(B) Schematic of mitochondrial delivery by photothermal nanoblade. A 3 μm inner diameter glass microcapillary pipette tip coated externally with titanium is positioned to lightly contact the cell surface. A 532 nm pulsed laser illumination triggers a cavitation bubble to open the membrane with coordinated delivery of donor mitochondria into a cell using a fluid pump.

(C) Representative confocal image of two foci of DsRed-labeled donor mitochondria from HEK293T cells in the cytosol of a single 143BTK− ρ0 cell whose endogenous mitochondria are stained with MitoTracker Green (upper left quadrant).

(D) Isolated MDA-MB-453 donor and 143BTK− parent cell mitochondria remain functional and coupled. Mean ± SD (n = 3).

(E) 2 weeks post-nanoblade delivery, donor MDA-MB-453 (and 143BTK− parent, not shown) mitochondria transferred into 143BTK− ρ0 recipient cells generated “rescue” clones that emerged in uridine-free dialyzed media (left). 143BTK− ρ0 control cells (or 143BTK− ρ0 cells that received 143BTK− ρ0 donor mitochondria, not shown) died and detached from the plate when grown in uridine-free dialyzed media (right).