Ventana staining system

After receiving institutional review board approval with a waiver of informed consent, we performed a retrospective chart and pathological review to identify patients of the senior author (MM) who underwent surgical neurolysis and resection of neuromas as part of surgical repair. The pathological review was performed to ensure tissue was available. Histology was performed at the discretion of the senior author and neuropathologist (CP); it included hematoxylin and eosin (H&E), trichrome, and laminin immunohistochemistry (IHC) stains. Five-micrometer, formalin-fixed, paraffin-embedded sections of neuroma resections from all patients were cut at regular intervals and mounted on glass sides. H&E and trichrome stains were prepared on all specimens except for Cases Two and Four. Immunohistochemical staining was performed using laminin antibodies (Leica Biosystems, Wetzlar‎, Germany) at a dilution of 1:100. Staining was performed using the avidin-biotinylated peroxidase complex (ABC) method on a Ventana Staining system (Ventana Medical Systems, Inc., Arizona, United States) and counterstained with hematoxylin.
The means (ranges) of patient ages and follow-up, along with the mechanism of injury and descriptive histopathological findings, were analyzed. Statistical analysis was not performed with this limited sample.

Immunohistochemical staining was performed on 5-micron cut sections of formalin-fixed, paraffin-embedded (FFPE) tissue blocks of all cases on Ventana staining system (Ventana Medical Systems, Tucson AZ, USA). All cases were stained with INI1 (monoclonal mouse antibody; 1:400; BD Bioscience, San Jose, CA, USA), Hep-par1 (mouse monoclonal antibody; 1:200; Dako, Santa Clara, CA, USA), Arginase (rabbit monoclonal antibody; 1:100; Sigma-Aldrich, St. Louis, MO, USA) and glypican-3 (mouse monoclonal antibody; ready to use; Sigma-Aldrich, St. Louis, MO, USA) antibodies.

4 μm sections were cut and used for H&E staining and immunohistochemistry (IHC) from each block. All slides were reviewed by two expert hematopathologists (LL, SL). Cases with insufficient or not optimal material were excluded from the study. The immunohistochemical analysis was performed with a panel of 25 antibodies commonly used in NHL and HL cases, using the Ventana staining system (Ventana, Tucson, AZ, USA).