For a total of 138 cases in Cohort A, Ki-67 immunohistochemistry was performed on formalin-fixed paraffin-embedded histopathological specimens. The immunohistochemistry was performed using an accredited methodology used in clinical routine practice with CONFIRM anti-Ki-67 (30-9) Rabbit Monoclonal Primary Antibody and stained with Ventana automated slide stainer (Ventana Medical Systems, Inc., USA). Analyses of lymph node and tonsils were included as positive controls and omission of the primary antibody served as negative control. Scoring was performed by calculating the percentage of positive cells (brown stained nuclei) in hotspot areas in at least 2000 cells.
Ventana automated slide stainer
Manufactured by Roche
Sourced in United States
The Ventana automated slide stainer is a laboratory equipment designed to automate the staining process for tissue samples. It is used to prepare slides for microscopic examination in various medical and research applications.
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Lab products found in correlation
5 protocols using ventana automated slide stainer
1
Ki-67 Immunohistochemistry for Tumor Analysis
2
FUCA-1 Expression in Tumor Samples
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Immunohistochemical analysis of FUCA-1 expression was performed on formalin-fixed, paraffin-embedded (FFPE) tumor sections using a rabbit polyclonal anti-FUCA-1 antibody (Proteintech Proteintech Group, Inc. Rosemont, IL, USA). Sections were stained using the Ventana automated slide stainer (Ventana Medical Systems, Tucson, Az, USA) with diluition 1: 100. Expression of FUCA-1 in tumor and normal cells was evaluated independently by two investigators (C.U. and F.B.) who were blinded to clinico-pathological data. Cytoplasmic staining of ≥25% of the tumor cells was considered positive, following the score: 0 <25%; + 25-50%; ++ 50-75%; +++ ≥75%. The possible association of FUCA-1 expression with clinico-pathological features was investigated by statistical analysis
3
Immunohistochemical Biomarker Detection
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ER and PR were detected by immunohistochemistry using CONFIRM™ mouse anti-ER antibody (clone 6F11) and CONFIRM™ mouse anti-PR antibody (clone 16) from Ventana Medical Systems. The staining was performed according to the manufacturer instructions using the Ventana® automated slide stainer (Ventana Medical Systems, S.A., Illkirch, France). HER2 was detected with the DAKO AO0485 polyclonal rabbit antibody also according to the guidelines provided by the manufacturer [35 (link)].
Ki67 was stained with the monoclonal mouse anti-human Ki67 clone MIB-1 (DAKO M7240) using the DAKO Link48 Auto Stainer protocol [36 (link)].
Ki67 was stained with the monoclonal mouse anti-human Ki67 clone MIB-1 (DAKO M7240) using the DAKO Link48 Auto Stainer protocol [36 (link)].
4
Evaluating Macrophage Presence in Tumor Samples
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In order to investigate the presence of macrophage in tumoral samples, CD68 expression was evaluated by IHC on FFPE tumor sections using mouse monoclonal anti-human antibody (Ventana Medical Systems, Inc., Tucson, AZ, USA). Sections were stained using the Ventana automated slide stainer (Ventana Medical Systems, Inc.). Expression of CD68 was evaluated independently by two of the authors (C.U. and F.B.) who were both blinded to the clinicopathologic data. Discrepancies between the two observers were discussed with a third pathologist (L.T.). The authors evaluated the CD68 expression in macrophages. The samples were evaluated as positive if staining was present. The intensity of the staining was evaluated in 5 high power fields (×40 magnification) and the ratio between the number of CD68 macrophages positive cells and the number of tumoral cells of the samples (i.e., 1:1 and 1:10) was used to create a graduated score: Low (1:100, 1:200), Medium (1:50, 1:20), and High (1:1, 1:2, 1:3, 1:5).
5
TUSC2 Expression in Thyroid Tumors
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Immunohistochemical analysis of TUSC2 expression was performed on formalin-fixed, paraffin-embedded (FFPE) tumor sections of 60 tumor cases, including 40 ATC and 20 PTC samples. As a control, thyroid tissues from 8 normal glands of patients who underwent surgery for head and neck tumors and 29 normal contralateral tissues were stained. Rabbit polyclonal anti-human TUSC2 antibody (Proteintech Group, Inc. Chicago, USA) was used (dilution 1:150). Sections were stained using a Ventana automated slide stainer (Ventana Medical Systems, Tucson, Az). TUSC2 expression in epithelial tumor and normal tissue cells was evaluated independently by two investigators (C.U. and F.B.) who were blinded to the clinicopathological data. Discrepancies between the two observers were discussed with a third pathologist (A.P.). For all cases, the percentage of positive cells per 5 high power fields (40X) was determined.
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