Journal: Reviews in Cardiovascular Medicine

Article Title: Targeting Tumor Necrosis Factor-α Mitigates Glucose Fluctuation-Induced Aortic Valve Fibrosis: Insights From Diabetic Rat Models

doi: 10.31083/RCM42804

Figure Lengend Snippet: Colocalization of CD3, CD68, and JAK1 in the aortic valves of diabetic rat models . (A,B) The coimmunostaining analysis of the HG and GF groups demonstrated the presence of CD3, CD68, and JAK1 proteins, indicating that JAK1 expression is primarily localized in T lymphocytes and macrophages. Notably, this expression pattern could be altered through the inhibition of TNF- α . Scale bar = 50 µm. DAPI, 4 ′ ,6-diamidino-2-phenylindole.

Article Snippet: In addition, the concentrated SABC-POD (Mouse/Rabbit IgG) kit (SA2010, BOSTER Biological Technology Co., Ltd., Wuhan, China), DyLight 488 conjugated AffiniPure goat anti-mouse IgG (H + L) (BA1126, BOSTER Biological Technology Co., Ltd., Wuhan, China), DyLight 594 conjugated AffiniPure goat anti-rabbit IgG (H + L) (BA1142, BOSTER Biological Technology Co., Ltd., Wuhan, China), ethylenediaminetetraacetic acid (EDTA) antigen retrieval solution (AR0023, BOSTER Biological Technology Co., Ltd., Wuhan, China), 4 ′ ,6-diamidino-2-phenylindole (DAPI) staining solution (AR1176, BOSTER Biological Technology Co., Ltd., Wuhan, China), human TNF- α enzyme-linked immunosorbent assay (ELISA) kit (EK0525, BOSTER Biological Technology Co., Ltd., Wuhan, China), and human TGF- β 1 ELISA kit (EK0513, BOSTER Biological Technology Co., Ltd., Wuhan, China) were obtained from Boster, China.

Techniques: Expressing, Inhibition

Journal: BMC Biology

Article Title: MATISSE: a method for improved single cell segmentation in imaging mass cytometry

doi: 10.1186/s12915-021-01043-y

Figure Lengend Snippet: Combining fluorescence microscopy with multiplex IMC data of colorectal tissue advances quality of single cell segmentation. a Cartoon describing MATISSE, a novel pipeline adding microscopic imaging to multiplex IMC analysis and downstream segmentation. In short: tissue sections on slides were stained using isotope-conjugated primary antibodies, DNA intercalator, and DAPI. The tissue was first scanned using a fluorescent microscope and then processed with IMC. Data produced by both techniques is aligned using the nuclear staining. Nuclear and membranous pixel probability maps are produced based on the fluorescent images and IMC data respectively. These probability maps are used to generate a segmentation map, where all detected cells are included. b Representative images of DNA intercalator on a colorectal tissue section analyzed by Ir193 labeling and IMC (left) or DAPI labeling and fluorescent microscopy (IF, right). c IMC-only (IMC) and MATISSE cell segmentation (MATISSE) were performed, and shown are the different predicted outlines on a representative image of Ir193 labeling. Arrows indicate areas with cell fragmentation. d Display of a large region of interest (ROI) showing an overlay of the predicted cell outlines (pink) upon IMC or MATISSE segmentation on a representative IMC image of DNA-Ir193 labeling of colorectal tissue. Highlighted in yellow is the approximate position of the basement membrane surrounding the epithelial monolayer. Scale bar 25 μm. e Cell density was calculated as the number of cells within a radius of 10 μM from the center of each single cell [ , ]. This number is displayed with a color code for each cell in the representative image

Article Snippet: We observed that incorporating fluorescent microscopy images based on DAPI nuclear staining into MATISSE workflow resulted in superior visual and signal intensity-based separation of nuclei in dense areas (Fig. b, Additional file : Fig S1B).

Techniques: Fluorescence, Microscopy, Multiplex Assay, Imaging, Staining, Produced, Labeling, Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Lyz2 -Cre-Mediated Genetic Deletion of Septin7 Reveals a Role of Septins in Macrophage Cytokinesis and Kras -Driven Tumorigenesis

doi: 10.3389/fcell.2021.795798

Figure Lengend Snippet: Effect of myeloid Septin7-deletion in myelopoiesis and BMDM proliferation. (A) Septin7-genotye analyses of bone marrow (BM) cells and BM-derived macrophages (BMDM) from the myeloid specific Septin7-KO mice show efficient deletion of the floxed allele. (B) Cell surface marker analysis of BM cells from Septin7 flox/flox :Lyz2-Cre and Septin7 wt/wt :Lyz2-Cre reveal no significant differences ( n = 3 mice each, p value > 0.3) in stem/progenitors (cKit + and Sca1 + ), monocytes (CD11b + and F4/80 + ) and granulocyte (Gr1 + and CD11b + ) populations. (C) Immunofluorescence analysis showing obligatory binucleation of Septin7 -KO BMDMs. DAPI is used for nuclear staining, WGA and Phalloidin (staining F-actin) are shown as counter stains. Septin7-negative cells display more than one nucleus, whereas Septin7-positive cells (stained green) display only one.

Article Snippet: DAPI for DNA staining was from Carl Roth (#6335.1).

Techniques: Derivative Assay, Marker, Immunofluorescence, Staining