3d sim image reconstruction  (Nikon)

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Quantitative Profiling of Alpha-Subunit of IL-3 Receptor on Single Acute Myeloid Leukemia Cells by Super-Resolution Imaging

doi: 10.31557/APJCP.2023.24.1.185

Figure Lengend Snippet: Quantitative Profiling and 3D Structure Reconstruction of CD123 on Molm13 and Molm13 R-5 Enriched Cells by SIM and Software Imaris. (A) Dispersion pattern of CD123 on the surface of single cell reconstructed from the raw SIM image. (B) Density and dispersion pattern of CD123 on Molm13 and Molm13-R5 single cells at angles of 60°, 120° and 240°. (C) Comparison of CD123 density on the surface of single cells for Molm13 and Molm13-R5 cells. Column sharing not same superscript letter differ significantly (P < 0.05).

Article Snippet: SIM image 3D reconstruction by Imaris 3D structure reconstruction and quantification of CD123 molecules was determined by software imaris as reported by us previously (Xi et al., 2019).

Techniques: Software, Dispersion, Comparison

Journal: bioRxiv

Article Title: Dynein acts to cluster glutamate receptors and traffic the PIP5 kinase, Skittles, to regulate postsynaptic membrane organization at the neuromuscular junction

doi: 10.1101/2021.09.27.462070

Figure Lengend Snippet: A-A”) The punctate distribution of dynein, while somewhat broader, is similar to that of the glutamate receptor subunit GluRIIC at the NMJ. A single optical section through the NMJ is shown. In the inset, many GluRIIC positive punctae overlap with dynein punctae and appear yellow (A’’) . An example of the GluRIIC localization in a control animal (B-B’) and in a dynein depleted animal (C-C’) used for the analyses in (D) . HRP labels the presynaptic side of the NMJ, and single optical sections through the NMJ are shown. D) Boxplots of the radius of iGluR spheres in control animals, and those depleted of dynein by RNAi. n=number of NMJ analyzed. Unpaired, one-tailed t-test results: ***, P<0.0001, **, P<0.005, *, P<0.05. E-E’’) Structured illumination microscopy images of the GluRIIC subunit and Bruchpilot (BRP), a presynaptic active zone component, in control animals and in animals depleted of dynein within the muscle (F-F’’) . G) Quantification of the number of BRP, active zone punctae apposed to a postsynaptic glutamate receptor cluster for control animals and those depleted of muscle dynein. n=number of NMJ analyzed, each point on the graph is an individual NMJ. Unpaired, two-tailed t-test results: ***, P<0.0001. Boxed areas are shown as insets. Scale, 10μm.

Article Snippet: Reconstructed 3D Structural Illumination Microscopy (SIM) images were analyzed using Imaris 9.5.1 (Bitplane, Belfast, UK).

Techniques: Control, One-tailed Test, Microscopy, Two Tailed Test

Journal: bioRxiv

Article Title: Dynein acts to cluster glutamate receptors and traffic the PIP5 kinase, Skittles, to regulate postsynaptic membrane organization at the neuromuscular junction

doi: 10.1101/2021.09.27.462070

Figure Lengend Snippet: Compared to controls (A-A’) , depletion of postsynaptic Sktl by RNAi results in increased cytoplasmic GluRIIC staining in the muscle and decreased extra synaptic GluRIIC punctae (B-B’) . Arrows indicated some of the extra synaptic punctae in (A,B) . C) Depletion of postsynaptic Sktl results in increased GluRIIC at the membrane compared to controls. Structured illumination microscopy was used to image and analyze GluRIIC clusters and their apposition to presynaptic BRP punctae. GluRIIC receptor cluster size, and their apposition to presynaptic Bruchpilot (BRP) punctae appear to be similar in controls (D-D’) and Sktl depleted animals (E-E’) . F) Quantification of the number of BRP, active zone, punctae apposed to a postsynaptic glutamate receptor cluster for control animals and those depleted of postsynaptic Sktl in comparison to the data shown in for dynein depletion, replicated again here for a side-by-side comparison. G-H’) Structured illumination microscopy was used to image glutamate receptor clusters and dynein together at NMJ to better determine the relationship between the two components. Two examples of wild type synaptic terminals are shown. GluRIIC labels glutamate receptor clusters, and Dhc64c labels the dynein heavy chain. In the merged image insets dynein often localizes to the center of the glutamate clusters (G’,H’) . I) A model for how dynein functions on the postsynaptic side of the NMJ to transport Sktl for local production of PIP 2 and organization of the postsynaptic spectrin cytoskeleton, and stabilization/organization of glutamate receptor clusters. n=number of NMJ analyzed, each point on the graph is an individual NMJ. Unpaired, two-tailed t-test results: ***, P<0.0001, **, P<0.005, *, P<0.05. Boxed areas are shown as insets. Scale, 10μm.

Article Snippet: Reconstructed 3D Structural Illumination Microscopy (SIM) images were analyzed using Imaris 9.5.1 (Bitplane, Belfast, UK).

Techniques: Staining, Membrane, Microscopy, Control, Comparison, Two Tailed Test