Journal: PLoS ONE
Article Title: Connexin 30 Expression and Frequency of Connexin Heterogeneity in Astrocyte Gap Junction Plaques Increase with Age in the Rat Retina
doi: 10.1371/journal.pone.0057038
Figure Lengend Snippet: Imaris snapshot images of confocal z-stack projections that visualise the entire depth of the retinal fibre layer where both astrocytes and blood vessels are situated. The overlapping immunohistochemistry represents connexin heterogeneity in gap junction plaques expressed by astrocytes and captured using high resolution three-dimensional projection, simulated fluorescence processing and orthogonal mode camera dialogue. The GFAP and GS isolectin B4 stain were omitted from these images to better illustrate the fluorescence of the connexins. A&C were taken in the central retinal region and B&D from a peripheral retinal region. A&C and B&D are a different angle of the same 3D analysis to show different connexin combinations as indicated by the arrows. Immunostaining of astrocyte connexins represents Cx26 (blue), Cx30 (red), Cx43 (green) and Cx45 (brown). Different colocalisation patterns of connexin proteins in diverse astrocyte connexin hemichannels were qualitatively analysed (1Cx protein, small arrows, 2Cx and 3Cx proteins, curved arrows, and 4Cx proteins, large arrowheads). All images are highly zoomed and rendered confocal z-slice projections. For qualitative purposes, displayed images were also constructed using automated scale bar, object frame (grid, tickmarks and box) and XYZ clipping plane (i.e. crops object below the plane). The set camera angle logarithmic functions were also applied to measure angle, elevation and azimuth parameters for each of the images represented. This was as follows: angle = −5.6521 (A), 4.4572 (B), −6.3247 (C), −0.0729 (D), elevation = 91.8453 (A), 88.6771 (B), 93.7761 (C), 90.0196 (D), azimuth = 119.9393 (A), −65.222 (B), 127.6513 (C), 93.3840 (D).
Article Snippet: The size of each Cx plaque was measured in voxels (X, Y and Z dimensions) and pixels (2-dimensions only) following acquisition using Imaris 3D-reconstruction image analysis and processing software (Bitplane Inc, Saint Paul, MN) version 7.0.
Techniques: Immunohistochemistry, Fluorescence, Staining, Immunostaining, Construct
![(a) Top panel. MicroCT views using <t>Fiji</t> <t>software</t> show tooth family unit development at resting (tooth family unit 7), initiation (tooth family unit 9), and growth (tooth family unit 8) phases by viewing a single plane of the <t>3D</t> dataset. Bottom panel. These same stages are shown from H&E staining of sectioned materials. Higher magnification views of the indicated areas are shown. (b) 3D visualization of teeth 7–12. (c) Tracing of tooth development at different hatching, 4 months and 1 year. f functional tooth, r replacement tooth, r1 replacement tooth 1, r2 replacement tooth 2, dl dental lamina. Panels A and C are from [11]](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_0952/pmc10880952/pmc10880952__nihms-1964883-f0004.jpg)
