Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Light-induced modulation of the mitochondrial respiratory chain activity: possibilities and limitations

doi: 10.1007/s00018-019-03321-z

Figure Lengend Snippet: IXplore SpinSR Olympus super-resolution microscopy reveals kinetics of mitochondria fragmentation upon HFLP laser irradiation. a Representative time-lapse super-resolution images of mitochondria in living Huh7 and Alexander cells stained with MitoTracker® red. Cells exposed to HFLP laser (398 and 650 nm; 50 µW) for indicated time periods. As a positive control, cells were treated with 20% ethanol for 15 min. Arrows show cites of mitochondria fission. b Kinetics of ΔmΦ upon HFLP laser irradiation. Huh7 and Alexander cells were labeled with 1 μM JC-1. Labeled cells were exposed to HFLP laser (398 and 650 nm; 50 µW) for indicated time periods. Quantitative analysis of ΔmΦ imaged by laser scanning confocal microscopy was performed utilizing LaserSharp 2000 software (BioRad). Data are expressed as mean ± SEM (n = 3), t = 0 timepoint served as control, ***p < 0.001. As a positive control, cells were treated with 20% ethanol. c Kinetics of mitochondrial ROS accumulation upon HFLP laser irradiation. Huh7 and Alexander cells were labeled with MitoTracker® red CM-H2XRos. Labeled cells were exposed to HFLP laser (398 and 650 nm; 50 µW) for indicated time periods. Quantitative analysis of MitoTracker® red CM-H2XRos fluorescence imaged by laser scanning confocal microscopy was performed utilizing LaserSharp 2000 software (BioRad). Data are expressed as mean ± SEM (n = 3), t = 0 timepoint served as control, ***p < 0.001. Non-irradiated cells treated with H2O2 (0.5 mM) were used as a ROS positive control

Article Snippet: Open in a separate window Fig. 3 IXplore SpinSR Olympus super-resolution microscopy reveals kinetics of mitochondria fragmentation upon HFLP laser irradiation. a Representative time-lapse super-resolution images of mitochondria in living Huh7 and Alexander cells stained with MitoTracker® red.

Techniques: Microscopy, Irradiation, Staining, Positive Control, Labeling, Confocal Microscopy, Software, Fluorescence

Journal: bioRxiv

Article Title: Spatial localization of CD16a at the human NK cell ADCC lytic synapse

doi: 10.1101/2024.08.09.605851

Figure Lengend Snippet: (A-C) The far-left column depicts a cartoon representation of activating conditions used to model CD16a mediated activation and displays representative images of NK-92 CD16a-SNAP confocal and MINFLUX data of immune synapses on coated glass. From left to right, images show confocal reference, MINFLUX data colored given through DBSCAN, a zoomed in region of MINFLUX localizations as marked in the overview, and trace centers (single localizations are colored in purple, and isolated pairs are in orange) overlayed on raw MINFLUX data (colored in light cyan). Conditions are glass coated with (A) α-CD16 and α-CD18 antibodies, (B) α-CD18 antibody, and (C) PLL. (D) Average Ripley’s H function (top) and individual region of interest (ROI) Ripley’s H function (bottom) of MINFLUX data. (E) Nearest neighbor analysis of all MINFLUX data. (F) Nearest neighbor analysis of isolated pairs of CD16a molecules. (G) The total number of isolated CD16a pairs observed in all ROIs. Scale bars in confocal images and raw MINFLUX renderings represent 500 nm and zoomed in region scale bars represent 50 nm.

Article Snippet: JM is an employee of the company Abberior Instruments America, which commercializes super-resolution microscopy systems, including MINFLUX.

Techniques: Activation Assay, Isolation

Journal: bioRxiv

Article Title: Spatial localization of CD16a at the human NK cell ADCC lytic synapse

doi: 10.1101/2024.08.09.605851

Figure Lengend Snippet: (A-B) The left column depicts a cartoon representation of activating conditions used to model CD16a mediated activation. Representative images of NK-92 CD16-SNAP confocal and MINFLUX data of immune synapse on SLBs. From left to right, images show confocal reference, MINFLUX data colored by ID given through DBSCAN, a zoomed in region of MINFLUX localizations as marked in the overview, and trace centers (single localizations are colored in purple, and isolated pairs are in orange) overlayed on raw MINFLUX data (colored in light cyan). SLBs contained HER2 and ICAM-1 (A) without Trastuzumab (-Traz) and (B) with Trastuzumab (+ Traz). (C) Average Ripley’s H function (top) and individual ROI Ripley’s H function (bottom) of MINFLUX data. (D) Nearest neighbor analysis of all MINFLUX data. (E) Nearest neighbor analysis of isolated pairs of CD16a molecules. Scale bars in confocal images and raw MINFLUX data represent 500 nm and zoomed in region scale bars represent 50 nm. MINFLUX microscopy data were collected across at least three independent experiments with a total of 12 cells analyzed.

Article Snippet: JM is an employee of the company Abberior Instruments America, which commercializes super-resolution microscopy systems, including MINFLUX.

Techniques: Activation Assay, Isolation, Microscopy

Journal: bioRxiv

Article Title: Spatial localization of CD16a at the human NK cell ADCC lytic synapse

doi: 10.1101/2024.08.09.605851

Figure Lengend Snippet: MINFLUX data of CD16a localization demonstrated that CD16a often is found in pairs with an intra-fluorophore distance of ∼17 nm apart. Shown is a cartoon representation of what this could look like in un-scaffolded CD16a (left) versus CD16a scaffolded by a homodimer such as pCD3ζ.

Article Snippet: JM is an employee of the company Abberior Instruments America, which commercializes super-resolution microscopy systems, including MINFLUX.

Techniques: