Journal: The Journal of Clinical Investigation

Article Title: Cyclooxygenase-2 in adipose tissue macrophages limits adipose tissue dysfunction in obese mice

doi: 10.1172/JCI152391

Figure Lengend Snippet: WT and myeloid COX-2 –/– mice were on the HFD for 11 weeks. ( A ) Hyperinsulinemic-euglycemic clamps determined more severe insulin resistance in myeloid COX-2 –/– mice, as less glucose infusion was needed to maintain a constant blood glucose ( n = 4). ( B – D ) Myeloid COX-2 –/– mice had increased plasma insulin levels at baseline and during clamp periods ( B ), decreased rates of glucose disappearance (RD) ( C ), and increased endogenous glucose production (EGP) ( D ) ( n = 4 and 5). ( E ) Myeloid COX-2 –/– mice had decreased glucose uptake, a marker of insulin resistance in adipose tissues (BAT, SAT, VAT), SM (gastrocnemius and soleus), and heart and brain ( n = 4 and 5). ( F ) Picrosirius red staining indicated more fibrosis in EF and IF in myeloid COX-2 –/– mice than WT mice ( n = 8). Scale bars: 100 μm. ( G ) Myeloid COX-2 –/– mice had decreased insulin-stimulated p-Akt in EF, SM, and liver, an indication of increased insulin resistance ( n = 4–6). ( H ) Quantitative p-Akt immunofluorescent staining showed insulin insensitivity in SM in myeloid COX-2 –/– mice ( n = 6). Scale bars: 100 μm. ( I ) Myeloid COX-2 –/– mice had lower Adipoq mRNA levels in EF and IF ( n = 6). Data are mean ± SEM. * P < 0.05, ** P < 0.01, analyzed using 2-way ANOVA followed by Tukey’s post hoc test for A , 2-way ANOVA followed by Bonferroni’s post hoc test for B – D , F , and I , and 2-tailed Student’s t test for E , G , and H . Brown, subcutaneous, and visceral adipose tissue (BAT, SAT, and VAT); EF, epididymal fat; IF inguinal fat; SM, skeletal muscle.

Article Snippet: Fat tissue fibrosis was determined using a Picrosirius red stain kit (Direct Red 80, 365548, Sigma-Aldrich).

Techniques: Marker, Staining

Journal: Cancers

Article Title: Assessment of Ovarian Tumor Growth in Wild-Type and Lumican-Deficient Mice: Insights Using Infrared Spectral Imaging, Histopathology, and Immunohistochemistry

doi: 10.3390/cancers13235950

Figure Lengend Snippet: Workflow showing the histology, the immunohistochemistry of formalin-fixed paraffin-embedded ID8 ovarian tumor sections, SHG imaging, Picrosirius red staining (polarized light), and analysis of FTIR images using common K-means clustering.

Article Snippet: To study ECM collagen organization, deparaffinized tissue sections were stained using the Abcam Picrosirius red stain kit (ab150681), according to the manufacturer’s instructions.

Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Imaging, Staining

Journal: Cancers

Article Title: Assessment of Ovarian Tumor Growth in Wild-Type and Lumican-Deficient Mice: Insights Using Infrared Spectral Imaging, Histopathology, and Immunohistochemistry

doi: 10.3390/cancers13235950

Figure Lengend Snippet: Analysis of collagen organization in ovarian tumor sections of wild-type and lumican-deficient mice. ( a , b ) Representative microphotographs of s.c. allograft sections stained with HES (top panel, original magnification 20×, scale bar 500 µm) in Lum +/+ ( a ) and Lum −/− mice ( b ); ( c – f ) Picrosirius red and SHG image analyses of ovarian allografts in tumors implanted in Lum +/+ mice ( c , d ) and in tumors from Lum −/− mice ( e , f ); ( c , e ) Collagen SHG images from ID8 ovarian tumors (original magnification 20×); ( d , f ) Ovarian tumor sections stained with Picrosirius red and viewed under widefield cross-polar optics (original magnification 20×, scale bar 50 µm). Birefringence of collagen fibers allows distinction between type I (red) and type III (green) collagens; ( g ) Analysis of collagen fibers intensity by SHG in tumors and healthy tissues present in each section (mean ± SD, ns: not significant); ( h ) Analysis of tumor ECM collagen organization from images derived from Gabor filtering and FFT, processed on Picrosirius red images (mean ± SD, ns: not significant); ( i ) Quantification on Picrosirius red stained sections of the relative distribution of red pixels (corresponding to type I collagen) in tumor ECM of Lum +/+ and Lum −/− sections (mean ± SD, * p < 0.05); ( j ) Quantification on Picrosirius red stained sections of the relative distribution of green pixels (corresponding to type III collagen) within tumors of Lum +/+ and Lum −/− sections (mean ± SD, * p < 0.05).

Article Snippet: To study ECM collagen organization, deparaffinized tissue sections were stained using the Abcam Picrosirius red stain kit (ab150681), according to the manufacturer’s instructions.

Techniques: Staining, Derivative Assay

Journal: Reproductive Sciences

Article Title: Cervix Stromal Cells and the Progesterone Receptor A Isoform Mediate Effects of Progesterone for Prepartum Remodeling

doi: 10.1177/1933719118820462

Figure Lengend Snippet: Top: Photomicrographs of progesterone receptor (PR) stained cells and counterstained cell nuclei (CN) in cervix sections from nonpregnant (NP), pregnant (days 15 and 18 postbreeding), and day of birth postpartum (PP) mice. Scale bar is 25 µm. Middle: Density of PR cells normalized to CN per area to account for variability in cell nuclei density due to heterogeneity of tissue morphology within and among sections in individuals, as well as within and among groups. Data are the mean ± SE (n = 5-21/group). Bottom: As inversely related to cross-linked collagen in the extracellular matrix,2 optical density of birefringence of picrosirius red-stained cervix sections cells was normalized to CN per area. Data are mean ± SE (n = 4-10/group; *P < .05 vs NP mice or **vs NP, day 15 and day 16 postbreeding groups by 1-way ANOVA). See Methods for details about staining and analyses. SE indicates standard error; ANOVA, analysis of variance.

Article Snippet: Additional sections were stained with a picrosirius red stain kit (PK-4000; Polysciences, Warrington, Pennsylvania) to assess extracellular collagen organization and counterstained with hematoxylin (SH26; Fisher Scientific, Asheville, NC) as described previously.

Techniques: Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: Protective Effect of Cimicifuga racemosa (L.) Nutt Extract on Oocyte and Follicle Toxicity Induced by Doxorubicin during In Vitro Culture of Mice Ovaries

doi: 10.3390/ani13010018

Figure Lengend Snippet: Representative images of collagen fibers labeled by Picrosirius red: ( A ) control group (DMEM + ); ( B ) DOXO (0.3 μg/mL); ( C ) CIMI (5 ng/mL) and ( D ) CIMI (5 ng/mL) + DOXO (0.3 μg/mL). Scale bar: 100 μm (400×).

Article Snippet: To assess collagen fibers of extracellular matrix of ovarian cortex, staining with Picrosirius Red (Abcam Kit) was performed following the methodology described by Rittié [ ] with modifications.

Techniques: Labeling, Control