Co-immunofluorescence (IF) was performed in a previously described cohort of 200 HGSC on tissue microarray (TMA)9 (link) and in a previously described cohort of 1000 HGSC (from the Canadian Ovarian Experimental Unified Resource)25 (link) on TMA (online supplemental table S1 ). Only cases that did not receive neoadjuvant chemotherapy were included for analyses. Slides were deparaffinized, rehydrated, demasked using a citrate buffer and incubated for 30 min with a protein blocking solution. Primary antibodies were co-incubated on the slides overnight at 4°C and secondary antibodies were incubated for 2 hours at room temperature (online supplemental table S2 ). Slides were then mounted with 4′,6-diamidino-2-phenylindole (DAPI). Sections were imaged at 20X magnification on VS-110 scanner (Olympus) and super images were imported into Visiomorph DP (Visiopharm). CK8-18 positivity was used to identify the epithelial area and where indicated α smooth muscle actin (SMA) positivity was used to identify the activated stromal area. Cores with epithelial or stromal areas representing less than 5% of the total tissue area were eliminated from survival analyses. Recommendations for Tumor Marker Prognostic Studies criteria were followed.26 (link)
Visiomorph dp
Manufactured by Visiopharm
Sourced in Denmark
Visiomorph DP is a digital pathology software platform developed by Visiopharm. It provides image analysis capabilities for processing and analyzing digital pathology slide images.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using visiomorph dp
1
Co-immunofluorescence Profiling of HGSC
2
Whole Slide Digital Analysis of Mammary Gland
3
Nasal Septum Histological Evaluation
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From each animal the nasal septum was carefully dissected out, subjected to overnight fixation in 4% buffered formaldehyde, dehydrated, and embedded in paraffin. From each septum, series of 3 μm sections (transverse section plan) were obtained from two depth levels, separated by 200 μm. Sets of sections from both depth levels were subjected to standard hematoxylin staining, Periodic Acid‐Schiff (PAS) staining (for goblet cell evaluation), FITC eosinophil granule staining 6 , 32 , and immunohistochemical (IHC) staining of T lymphocytes. Sections for IHC were subject to Heat‐Induced Epitope Retrieval (HIER, PT‐link machine, Dako Cytomation, Denmark) prior to automated IHC staining (Dako Autostainer Robot, Dako) using a mouse anti‐guinea pig T cell monoclonal antibody (MCA751, clone CT5, AbD Serotec). For quantification, stained sections were digitalized in an Olympus VS‐120 slide scanner. The epithelial area was outlined my manual cursor tracing the histochemical staining for eosinophils, T cells, and PAS were automatically quantified by computerized image analysis (VisiomorphDP, Visiopharm, Denmark).
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