deconvolution algorithm deltavision softworx image analysis program  (Softworx Inc)

Journal: The Journal of Biological Chemistry

Article Title: Multi-modal mechanisms of the metastasis suppressor, NDRG1: Inhibition of WNT/β-catenin signaling by stabilization of protein kinase Cα

doi: 10.1016/j.jbc.2024.107417

Figure Lengend Snippet: Triple co-localization of β-catenin, NDRG1, and PKCα demonstrates the formation of a possible metabolon, decreasing β-catenin levels and nuclear localization. A – H , NDRG1 overexpression in PANC-1 cells significantly increases the co-localization between β-catenin, NDRG1, and PKCα. A and B , VC and NDRG1 overexpressing PANC-1 cells were incubated for 24 h/37 °C in the presence and absence of WNT3a (100 ng/ml). The cells were then examined for β-catenin ( green ), NDRG1 ( red ), and PKCα ( blue ) expression and triple co-localization ( white ) using confocal immunofluorescence microscopy. Studies were performed using a 100× objective at the same acquisition setting with Olympus Fluoview FV3000 software. Images were digitally magnified for better demonstration of the possible co-localization. The scale bar = 3 μm for all images except for the inset images, where the scale bar = 9 μm. C and D , deconvolution analysis was then implemented on the images in ( A and B ) using Olympus CellSens imaging software. White arrows demonstrate triple co-localization and possible formation of the metabolon. The scale bar = 3 μm for all deconvolution images except for the inset images, where the scale bar = 1.8 μm. In all studies, the images are representative of three experiments, and the quantitative analysis of the pixel intensities is shown for ( E ) β-catenin; ( F ) NDRG1; ( G ) PKCα; and ( H ) Co-localization between β-catenin, NDRG1, and PKCα. Quantitative analyses were performed using ImageJ software and were presented as the mean ± SD ( n = 3). Analysis of pixel intensity and co-localization were performed using 24 cells. Statistical significance is denoted as ∗∗ p < 0.01 and ∗∗∗ p < 0.001 comparing NDRG1 overexpressing PANC-1 cells in the presence and absence of WNT3a to VC cells in the absence of WNT3a; or ### p < 0.001 comparing NDRG1 overexpressing PANC-1 cells to VC cells in the presence of WNT3a.

Article Snippet: The images of visualized cells were then examined using Olympus Fluoview software, and in some studies, images were processed for deconvolution analysis with CellSens software (Olympus) to improve contrast and image resolution.

Techniques: Over Expression, Incubation, Expressing, Immunofluorescence, Microscopy, Software, Imaging

Journal: Cancers

Article Title: Lysyl-Oxidase Dependent Extracellular Matrix Stiffness in Hodgkin Lymphomas: Mechanical and Topographical Evidence

doi: 10.3390/cancers14010259

Figure Lengend Snippet: Digital pathology imaging and measurement of lymph node stroma in HL and NHL. Slides of hematoxilin-eosin (HE, representative cases in Panel ( A )) or Mallory trichrome staining (representative cases in Panel ( B )) from LN sections of patients with HL ( n = 11), FL1-2 ( n = 6), FL3A ( n = 8), DLBCL ( n = 6), compared with 9 LDN, were subjected to Digital Pathology Imaging. Panel ( C ). HE sections were analyzed with the Genie software (right images in Panel ( A )) and the percentage area of stromal compartment per slide was calculated, adjusted to total tissue area in the image analyzed. Data are expressed as a percentage of green pseudocolor (stromal) areas and are the mean ± SEM of the indicated number of cases for each histological type. * p < 0.05 vs. LDN, FL1-2 and FL3A. Panel ( D ): digital images of Mallory trichrome stained sections were analyzed with an automated image analysis algorithm for color deconvolution (right images in Panel ( B )) and collagen areas quantified by the Image Scope software. Data are expressed as percentage of blue (collagen) areas and are the mean ± SEM of the indicated number of cases for each histological type. * p < 0.005 vs. LDN, FL1-2 and FL3A.

Article Snippet: Layers of each virtual slide annotation were created manually, selecting the whole area, and analyzed with the automated image analysis algorithm color deconvolution (Aperio software version 9.1, Aperio Technologies, Nussloch GmbH, Wetzlar, Germany).

Techniques: Imaging, Staining, Software