mitochondria matrix targeted memerald  (Addgene inc)

Journal: Nature Communications

Article Title: FidlTrack: high-fidelity structure-aware single particle tracking resolves intracellular molecular motion in organelles sensing APP processing

doi: 10.1038/s41467-026-69067-y

Figure Lengend Snippet: a TIRF images of a frame from simultaneously acquired single-molecules (left) and the associated organelle geometries (right) for a mitochondria matrix targeted HaloTag probe and mNeonGreen marker (top), the mitochondria membrane protein TOMM20 and mitochondria matrix targeted mEmerald (middle), and the ER membrane protein SEC61B and ER lumen targeted mEmerald (bottom) in COS-7 cells. b Masks extracted via AI-assisted image processing from the first and last frame of the recordings for the mitochondria matrix targeted probe (top), TOMM20 (middle) and SEC61B (bottom) recordings presented in ( a ). c Stability maps, representing for each pixel the percentage of total frames in which it is part of a mask, for the mitochondria matrix targeted probe (top), TOMM20 (middle) and SEC61B (bottom) recordings presented in ( a ). d Bar plot presenting the fraction of recovered spots relative to the quantity recovered from the entire stack of masks when using the mask of either the first or last frame to perform structure-aware tracking in the different recordings presented in ( a ). e Reconstructed trajectories (individually colour-coded) from the datasets presented in ( a ) using conventional tracking (not structure-aware, top, left), structure-aware tracking using the mask from the first frame (top, right) or the whole stack of masks (bottom, left). f Plot of the displacement distributions extracted from the trajectories presented in ( e ) fitted to a mixture of two Rayleigh distributions (dashed red lines, see Methods section “Fitting of displacement lengths distributions”). g Plot of the pooled displacement distributions for structure-aware ambiguity-removed tracking of SEC61B::Halotag in COS-7 cells after 4 h of BSA or Oleic Acid treatment at 400 µM. Pooling was done over n =7/6 recordings for BSA and oleic acid, respectively, the reported statistics correspond to a two-tailed Kolmogorov–Smirnov test p < 2.2251e-308. h Bar plot of the Kolmogorov–Smirnov statistics from ( g ) for either conventional or structure-aware tracking and with or without ambiguity removal. i Temporally colour-coded mitochondria positions over 700 frames (left), trajectory of the centre of the pointed mitochondria (right, top) and the raw (black) and mitochondrion motion-corrected trajectory (red) of a TOMM20 receptor moving at the surface of the pointed mitochondrion (right, bottom). Source data are provided as a Source Data file.

Article Snippet: 827 , mEmeraldMito , mEmerald Mito , Mitochondria matrix targeted mEmerald , Fig. , Addgene #54160.

Techniques: Marker, Membrane, Two Tailed Test

Journal: Nature Communications

Article Title: FidlTrack: high-fidelity structure-aware single particle tracking resolves intracellular molecular motion in organelles sensing APP processing

doi: 10.1038/s41467-026-69067-y

Figure Lengend Snippet: a Confocal images of a COS-7 cell stably expressing the ER marker mEmerald ER , an ER-targeted anti-alfatag intrabody fused to an HaloTag (stained with Halo-JFX650), an Amyloid Precursor Protein containing the Alfatag peptide and fused to a SnapTag (stained with Snap-TMR) and the merged image showing the near perfect correlation of the three signals. b Principle of the intrabody status detection method at the single-molecule level. Unbound intrabodies have a relatively fast luminal dynamic while bound intrabodies acquire the slower membrane dynamics of their APP target (Created with Biorender ). c TIRF images of ER structures (top) and associated FidlTrack reconstructed single-particle trajectories (colour-coded by the average trajectory displacement length of each trajectory) for cells expressing: the intrabody together with the ER-retained Snap-tagged APP (stained with Snap-PA-JF646, left), the ER-retained intrabody alone (stained with Halo-PA-JF646) (middle), or the intrabody (stained with Halo-PA-JF646) together with the APP construct (right). d Distributions of the pooled averaged trajectory displacement lengths for the unbound intrabody (red), APP (blue) and intrabody + APP (purple) recordings. The dashed lines correspond to fits of the distributions to single-components Gaussian models. The pooling is over 5 recordings for each condition. e Distributions of the averaged trajectory displacement lengths for 5 individual recordings of the intrabody in the presence of APP. The dashed lines correspond to fits of the distributions to two-component Gaussian models. f Average trajectory displacement length parameter (fitted value ± 95% CI) of each component extracted from the fits of the different distributions presented in ( e ). The dashed lines correspond to the average trajectory displacement length parameters of the single-component Gaussian fits of the APP (blue) and intrabody-only (red) recordings from ( d ). g Associated proportion of bound and unbound intrabody populations for the fits presented in ( e ) (see Methods section “Binding status extraction from intrabody trajectories”). h Principle of the procedure for unmixing APP-bound and unbound intrabody trajectories. i Unmixing of APP vs unbound intrabodies on a recording, from left to right: ER structure of a cell expressing both the intrabody and APP, recorded trajectories, APP trajectories and unbound trajectories, colour coded by individual trajectory. Source data are provided as a Source Data file.

Article Snippet: 827 , mEmeraldMito , mEmerald Mito , Mitochondria matrix targeted mEmerald , Fig. , Addgene #54160.

Techniques: Stable Transfection, Expressing, Marker, Staining, Membrane, Single Particle, Construct, Binding Assay, Extraction