Envision flex high ph visualization system

Tissue arrays were constructed using the 124 paired CRC tissues and metastatic tissues. Immunohistochemical staining was performed on 4-mm sections of paraffin-embedded tissues to determine the expression level of proteins. In brief, the slides were incubated in the first antibody (CST) diluted 1:200 at 4 °C overnight. The subsequent steps were performed using the EnVision FLEX High pH visualization system according to the manufacturer’s instructions (Dako).

Tissue samples were fixed in 10 % neutral-buffered formalin, then processed and embedded in paraffin. Sections were cut at 5 μm for hematoxylin and eosin (Richard-Allan Scientific, Kalamazoo, MI, USA) staining and immunohistochemistry.
Immunohistochemical evaluation was performed using monoclonal antibodies specific for α-smooth muscle actin (α-SMA), an indicator of myofibroblast presence (Clone 1A4, DAKO, Glostrup, Denmark) and for CD68 (Clone KP1, DAKO, Glostrup, Denmark [antibody recognizes a 110-kDa glycoprotein expressed on monocytes and macrophages]). All immunohistochemistry was performed using the EnVision™ FLEX High pH visualization system (DAKO, Glostrup, Denmark).
General characteristics of the histopathology of implant capsules with different Baker scores were assessed visually by review of hematoxylin and eosin-stained capsule samples. Capsules were classified into four categories: (1) dense collagen, acellular or low cellular content (example Fig. 4a), (2) dense collagen, moderate to high cellular content (example Fig. 4b), (3) synovial metaplasia (example Fig. 4c, d), or (4) loosely packed collagen (example Fig. 4e, f).

Tissue arrays were conducted using 124 CRC tissues and 46 paired metastatic tissues. IHC staining was performed on 4 mm sections of paraffin embedded tissues to determine the expression level of proteins. In brief, the slides were incubated in first antibody diluted 1:50 to 1:200 at 4 °C overnight. The subsequent steps were performed using the EnVision FLEX High pH visualization system according to the manufacturer’s instructions (Dako, Denmark).