Journal: Scientific Reports

Article Title: A new model of sperm nuclear architecture following assessment of the organization of centromeres and telomeres in three-dimensions

doi: 10.1038/srep41585

Figure Lengend Snippet: Panels a–d display the 3D localization of telomeres in four representative sperm cells. Nuclei are stained blue with DAPI, with telomere signals shown in green. Nuclei were captured with a Z step of 0.2 μm, with a minimum of 92 sections per nuclei using wide-field microscopy (Deltavision) and subjected to deconvolution (SoftWoRx). The first image on the left side of panels a–d, shows the raw image following deconvolution. 3D models were rendered with Imaris software (Bitplane, Switzerland, the second image in panels a–d, display the formation of 3D models with rendered telomeres (merged composite). The third image subsequently shows the same models with the DAPI partially removed (DAPI plane clipped) to better observe the 3D radial distribution of telomeres throughout all regions of the nucleus. The fourth and fifth images show the same DAPI clipped models rotated along the X-axis to further depict the 3D radial organization. Scale bar = 3 μm. Panel e displays a single sperm cell following 3D modeling that clearly depicts the sperm tail that is utilized to evaluate the longitudinal organization of telomeres with full DAPI, partially clipped DAPI and no DAPI. This clearly displays that sperm telomeres are longitudinally localized throughout the sperm nucleus from the sperm head to the tail). Scale bar = 4 μm.

Article Snippet: 3D reconstruction of the captured images (as 3D stacks) from SoftWoRx were rendered in 3D and analyzed using the Imaris software (V.7.6.3 Bitplane – Zurich, Switzerland) by converting images to 32-bit float images.

Techniques: Staining, Microscopy, Software

Journal: Scientific Reports

Article Title: A new model of sperm nuclear architecture following assessment of the organization of centromeres and telomeres in three-dimensions

doi: 10.1038/srep41585

Figure Lengend Snippet: Panels a–e display the 3D localization of centromeres in two representative sperm cells. Nuclei were captured with a Z step of 0.2 μm, with a minimum of 92 sections per nuclei using wide-field microscopy (Deltavision) and subjected to deconvolution (SoftWoRx). The upper panel shows the raw image following deconvolution. The middle panel shows the 3D rendered models using the Imaris software, for the same cells. Nuclei are stained with DAPI (blue in the raw image; pseudocolored gray in the 3D rendered models); the pancentromeric probe staining all centromeres are red ( a ); nuclear organizing regions (NOR) are green ( b ); and centromeres 1, 5, and 19 are aqua ( c ). Image d shows the merged image of all investigated loci, and image e is the same merged image rotated 180°, showing the backside of the cell. The bottom panel, shows the same two sperm cells with the DAPI plane partially removed (clipped) and rotated along the X-axis to better observe the radial 3D distribution of centromeres and NOR throughout the sperm nucleus. Scale bar = 5 μm.

Article Snippet: 3D reconstruction of the captured images (as 3D stacks) from SoftWoRx were rendered in 3D and analyzed using the Imaris software (V.7.6.3 Bitplane – Zurich, Switzerland) by converting images to 32-bit float images.

Techniques: Microscopy, Software, Staining

Journal: bioRxiv

Article Title: Multi-color 4D superresolution light-sheet microscopy reveals organelle interactions at isotropic 100-nm resolution and sub-second timescales

doi: 10.1101/2021.05.09.443230

Figure Lengend Snippet: a , Dual-color volume rendering (top panel) of the ER (tagged with EGFP-Sec61β) and mito Ma (tagged with Cox4-mKate2) in a whole live U2OS cell. The lower panels show the y-z and x-z MIPs along the sectional planes in the top panel. Scale bar, 5 μm. b , Illustrative ROI, shown as one x-y MIP and three corresponding 3D renderings from different perspectives, revealing the real spatial relationships and contacts between the ER and mitochondria otherwise hardly visible in x-y views (top row). Scale bar, 500 nm. c , Illustrative ROI, shown as x-y, y-z, and x-z MIPs and a 3D rendering, revealing the ER wrapping percentages (the white numbers) at the contact sites otherwise indiscernible in the lateral images. Scale bar, 500 nm. d, e , Mitochondrial morphology at the ER-mitochondria contact sites with similar or different ER wrapping percentages. Scale bar, 200 nm. The white or blue arrows in b, d, e indicate the ER-mitochondria contact sites. f, g , Time-lapse dual-color volume renderings showing the dynamic processes of ER-mediated mitochondrial division. The images in the white boxes show the raw reconstructed images of the corresponding y-z sectional planes. The 2D and 3D models show the percentages of the ER wrapping around the mitochondrial circumference (the contact sites marked in red). h , Quantitative statistics of the ER wrapping percentage at the division sites during the ER-mediated mitochondrial division events (n=23). i , Time-lapse dual-color volume renderings showing the dynamic process involving the cooperation of the ER and one mitochondrion (the white arrows) in the division of another mitochondrion (the blue and yellow arrows). The zoomed-in view shows the contact site where the ER occurs in conjunction with the two mitochondria. Scale bar, 200 nm.

Article Snippet: All the others 3D rendering images were generated by loading the time-lapse image stacks into Amira or Imaris software.

Techniques: