image processing and 3d reconstruction  (MathWorks Inc)

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

Journal: Microsystems & Nanoengineering

Article Title: Manipulation of single cells via a Stereo Acoustic Streaming Tunnel (SteAST)

doi: 10.1038/s41378-022-00424-9

Figure Lengend Snippet: a SteAS-based trapping of a single fluorescent PS particle (5 μm). When the power was applied, particles are trapped and suspended in the microchannel. b When the power is turned off, the particles settle to the surface. c A suspended particle is trapped and rotated in the AST under the combined effect of radiation force and drag force. The stacked image sequences (6 images, 27 ms apart) are captured at approximately 10 mW. The green dotted circle represents the range of trapping points. The red arrows show the direction of particle rotation. The yellow dotted rectangle shows the position of the UHF device. d Pattern of individual cells based on the AST. HeLa cells stained with calcein-AM (green) are used to represent the size and profile of the trapping point. The white arrow represents the direction of lateral flow, the red arrows show the trajectories of individual cells in SteAS, the white dotted pentagon is the position of the UHF device, and the blue curved rectangles represent the profile of the AST. e A single cluster trapped in the AST. The cells are highlighted by pseudocolor (red). The detailed images show the images before and after image processing. The rotation axis and direction are represented by a blue dotted line and a blue arrow, respectively. f The result of 3D reconstruction.

Article Snippet: The image processing and 3D reconstruction were achieved by MATLAB software (USA).

Techniques: Staining