Journal: bioRxiv

Article Title: Mapping brain-wide excitatory projectome of primate prefrontal cortex at submicron resolution: relevance to diffusion tractography

doi: 10.1101/2021.09.13.460040

Figure Lengend Snippet: The STP tomography was integrated with a two-photon microscope and a vibratome, both of which were computer controlled and fully automated. The traveling range of XYZ stage was customized to 50 by 60 by 70 mm to cover the entire macaque brain.

Article Snippet: High resolution fluorescent images were acquired with a confocal laser microscope Nikon TiE (Nikon, Tokyo, Japan) with a Plan Fluo 40× Oil DIC N2 objective (N.A.

Techniques: Tomography, Microscopy

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Localization of Angiopoietin-1 and Tie2 Immunoreactivity in Rodent Ependyma and Adjacent Blood Vessels Suggests Functional Relationships

doi: 10.1369/jhc.2009.954610

Figure Lengend Snippet: Primary antibodies and lectin used in tissue section labeling, immunoprecipitation, and Western blotting

Article Snippet: Tie2 , Santa Cruz Biotechnology , sc-9062 , Rabbit , Internal aa 234–533 (human) , 1:25 , Cy3.

Techniques: Labeling, Immunoprecipitation, Western Blot, Immunolabeling, Amplification, Plasmid Preparation

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Localization of Angiopoietin-1 and Tie2 Immunoreactivity in Rodent Ependyma and Adjacent Blood Vessels Suggests Functional Relationships

doi: 10.1369/jhc.2009.954610

Figure Lengend Snippet: Relationships of ependymal and vascular Angpt1 IR and tyrosine kinase 2 (Tie2) IR in normal adult female rats (confocal microscopy). (A) Angpt1 IR in aqueductal ependyma; (B) tomato lectin (T. Lect.)–labeled ependyma and a peri-ependymal blood vessel (bv); (C) merge of A,B and Hoechst nuclear stain (blue color) showing Angpt1-immunoreactive processes between the ependyma and an adjacent blood vessel. (D) Angpt1 IR in aqueductal ependyma; (E) Tie2 IR in a membranous pattern in the ependymal layer and in an adjacent blood vessel (bv). Insert: digital magnification of a portion of the ependyma showing concentration of Tie2 IR (red) on the lumenal surface and between adjacent ependymal cells in a Hoechst (blue)-stained section; (F) merge of D,E and Hoechst nuclear stain (blue color) showing Angpt1-immunoreactive processes (arrow) directed toward the Tie2-immunoreactive blood vessel (bv) and close association of membranous Tie2 IR and cytoplasmic Angpt1 IR in the cuboidal ependyma. (G) Angpt1-immunoreactive process; (H) Tie2 IR in a peri-ependymal blood vessel (bv); (I) merge of G,H and Hoechst nuclear stain (blue color) showing the close relationship between the Angpt1-immunoreactive process (arrow) and the Tie2-immunoreactive blood vessel (bv). (J) Angpt1 IR in a blood vessel; (K) Tie2 IR (arrow); (L) merge of J,K and Hoechst nuclear stain (blue color) showing the association between vascular Angpt1 IR and Tie2 IR. Note: Panels E and F were digitally postprocessed by linearly attenuating the intensity of the red signal using the “Image-Adjust-Levels” command in Adobe Photoshop software (Adobe Systems). Asterisk indicates aqueduct lumen. Bars: A–F = 25 μm; G–L = 10 μm.

Article Snippet: Tie2 , Santa Cruz Biotechnology , sc-9062 , Rabbit , Internal aa 234–533 (human) , 1:25 , Cy3.

Techniques: Confocal Microscopy, Labeling, Staining, Concentration Assay, Software

Journal: bioRxiv

Article Title: Solvatochromic reporter to image plasma membrane order leaflet by leaflet reveals a highly asymmetric bilayer locally modulated by transbilayer interactions

doi: 10.1101/2024.07.23.604763

Figure Lengend Snippet: (a) Confocal images of equatorial plane of cells labelled with 1uM Laurdan, NR12S or C3L showing the sub-cellular distribution of the respective probes. (b) Schematic showing the BSA back extraction assay used for extracting the outer leaflet population of probe in adherent cells. (c-f) Confocal images (c, e) collected from the bottom membrane plane of untreated cells (left panels) or cells treated with 2mM NEM to inhibit flippase activity (c) or 10uM Ionomycin to activate scramblase activity (e), before (top) and after (bottom) back extraction by BSA. Graphs (d, f) show the extent of incorporation (-) and remainder (+) of C3L after extraction with BSA. Each dataset is normalized to the mean intensity of untreated cells labelled with C3L. Data were obtained from from at least 40 cells. (g) Graph shows the time course of fluorescence intensity of C3L remaining at cell surface as a function of exposure to BSA added at t=0 mins (obtained using a CSU22 spinning disk confocal) in untreated cells (-), NEM treated cells (--) and NEM+Ionomycin treated cells (-.-). Each dataset is normalized to its mean starting intensity at t=0mins. Lines indicate fits to single exponential decay where k is the rate constant for the decay in fraction of C3L at the membrane after adding BSA, C i is the BSA resistant fraction after back-extraction. Data were obtained from atleast 20 cells in each condition. (h) Table shows rate constant (k) and BSA resistant fraction (C i ) obtained from the fit to a single exponential decay shown in panel g. All data is plotted as mean±s.d. p values determined from unpaired t test where ** indicates significance indicated by p<0.005. Scale=10µm.

Article Snippet: Live confocal imaging was done on a CSU W1 SORA (or where indicated a CSU22 mounted on Nikon TiE microscope with Andor iXon 897 cameras) spinning disk mounted on a Nikon Eclipse Ti2 microscope equipped with two Andor iXon 888 cameras.

Techniques: Extraction, Membrane, Activity Assay, Fluorescence