Diva or borg decloaking solution

Biopsy samples for histopathology and immunohistochemistry were immediately placed in formalin and processed by routine histologic methods and assessed by a board certified pathologist. Immunohistochemistry was performed on formalin fixed, paraffin embedded specimens following antigen retrieval with Diva or Borg decloaking solution (Biocare Medical). Adjacent 10 μm sections were stained for CD3⁺ (rabbit anti-human CD3, DAKO, USA; biotinylated goat anti-rabbit IgG, Vector Labs, USA; streptavidin, Biogenix; DAB, DAKO, USA), or FoxP3⁺ (rat anti-FoxP3, Ebioscience, USA; rat-on-mouse HRP probe/polymer, Biocare, USA; AEC, Biocare, USA). CD3⁺ and FoxP3⁺ cells in the dermis and epidermis were quantified from images captured at 400x magnification using a DP25 camera and BX53 microscope (Olympus, USA).

Biopsy samples were immediately placed in formalin and processed by routine histologic methods, followed by hematoxylin and eosin staining and assessment by a board certified pathologist. Immunohistochemistry was performed on formalin fixed, paraffin embedded specimens following antigen recovery with Diva or Borg decloaking solution (Biocare Medical, Concord, CA). Adjacent 10-μm sections were stained for CD3⁺ (rabbit anti-human CD3 [DAKO, Carpinteria, CA]; biotinylated goat anti-rabbit IgG [Vector Labs, Burlingame, CA]; streptavidin [Biogenex, Fremont, CA]; DAB [DAKO]), or FoxP3⁺ (rat anti-FoxP3 [eBioscience]; rat-on-mouse horseradish peroxidase probe/polymer [Biocare, Concord, CA]; AEC [Biocare]). CD3⁺ and FoxP3⁺ cells in the dermis and epidermis were quantified from images captured at 400× magnification using a DP25 camera and BX53 microscope (Olympus, Center Valley, PA).