Anti glial fibrillary acidic protein antibody

Each ERM specimen was collected during vitrectomy surgery for ERM. The tissue was immediately fixed with 4% paraformaldehyde and cryoprotected; then, 10-lm sections were obtained, as previously described. 33, 34 The sections were stained with hematoxylin and eosin, or immunostained with anti-alpha smooth muscle actin (aSMA) antibody (1:200; Sigma-Aldrich, St. Louis, MO, USA), anti-glial fibrillary acidic protein (GFAP) antibody (1:400; Cell Signaling Technology, Danvers, MA, USA), and anti-collagen type I antibody (1:200; Rockland Immunochemicals, Inc., Limerick, PA, USA); they were then visualized with Alexa 488-and Alexa 594-conjugated antibodies (1:1000; Invitrogen, Carlsbad, CA, USA), as well as 4 0 , 6diamidino-2-phenylindole (DAPI; Invitrogen). Images were acquired with a BioImaging Navigator fluorescence microscope (BZ-9000; Keyence, Osaka, Japan). This study was conducted in accordance with the guidelines of the Declaration of Helsinki; the protocol was registered within the UMIN Clinical Trial Registry (registered number UMIN000024553) and approved by the Nagoya University Hospital Ethics Review Board. Written informed consent was obtained from all participating patients.