CellLight Mitochondria-GFP, BacMam 2.0, provides an easy way to label mitochondria with green fluorescent protein (GFP) in live cells. Simply add the reagent to your cells, incubate overnight, and the cells are ready to image in the morning.

In this chapter, we detail the use of advanced confocal microscopy and mitochondrial matrix-targeted photoactivatable green fluorescent protein (mito-PAGFP) for the investigation of mitochondrial dynamics. We focus on direct visualization and quantification of mitochondrial fusion and mitochondrial network complexity in living mammalian cells.

PAGFP can be targeted to different parts of the mitochondrion in order to examine the mixing properties that underlie fusion events between these organelles. Once targeted, spatially precise photoconversion can be used to induce GFP fluorescence.

Sep 9, 2019 · Here we engineered MitoTag mice, which express a Cre recombinase-dependent green fluorescent protein targeted to the outer mitochondrial membrane, and developed an isolation approach to profile...

Because of the finite mitochondrial size, the 82– 91% recovery indicates that >90% of the GFP is mobile. The prompt and nearly complete fluorescence recovery was an unexpected observation, and suggested relatively free and rapid diffusion of GFP in the mitochondrial matrix.

Dec 11, 2017 · Here we report a split-GFP-based contact site sensor (SPLICS) engineered to fluoresce when organelles are in proximity. Two SPLICS versions efficiently measured narrow (8–10 nm) and wide (40–50...

Jul 13, 2020 · The Bi-Genomic Mitochondrial-Split-GFP, where both fragments of the Split-GFP are expressed by separated translation machineries, shuts off cytosolic fluorescence of dual-localized proteins, allowing visualization of their mitochondrial echoforms.

Mar 6, 2025 · The expression of either Halo–Miro1 or Halo–Miro2 stabilized Myo19 and rescued its expression by recruiting it to the mitochondria as verified by the GFP signal after transient transfection of GFP–Myo19 . Because upon downregulation of Miro, Myo19 is downregulated concomitantly, we had to take a different approach for analyzing whether ...

Jul 26, 2024 · We made use of a recently developed Bi-genomic split-GFP assay (BiG Mito-Split-GFP, hereafter shortened as BiG Mito-Split) where a larger fragment of GFP, GFP 1-10 is encoded in the mitochondrial DNA, and the small fragment GFP 11 is fused with a nuclear-encoded protein (Bader et al, 2020).

3 days ago · In most eukaryotes, mitochondrial organelles contain their own genome, usually circular, which is the remnant of the genome of the ancestral bacterial endosymbiont that gave rise to modern mitochondria. Mitochondrial genomes are dramatically reduced in their gene content due to the process of endosymbiotic gene transfer to the nucleus; as a result most mitochondrial proteins are encoded in the ...