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To report histopathologic analysis of corneas after corneal collagen crosslinking (CXL) for the treatment of keratoconus. Prospective, noncomparative, consecutive case series. Sixteen eyes of 13 patients with keratoconus who had undergone CXL were evaluated histologically with the aid of methylene blue-stained, Masson trichrome-stained, and immunofluorescent (IF) stained corneas. Color photographs of the corneas were also taken. Tissue staining was reviewed by 2 independent observers. The staining intensity of the basement membrane zone (BMZ) was assessed using a graded scale (0 to 4) and categorized as no staining, mild staining, moderate staining, or severe staining. Any staining that occurred beyond the endothelium was not included in the grading. The staining intensity of the BMZ in the crosslinking and non-crosslinking areas was compared. All 16 corneas showed the presence of IF staining, which was concentrated within the stromal layers of the cornea. No staining was observed within the endothelium, Descemet membrane, or Bowman layer. Evaluation of the IF staining intensity of the BMZ revealed that the intensity was lower in the stromal layers of the crosslinking area, indicating loss of IF staining in these areas. Several IF-stained specimens showed hyaline deposits in the stromal areas of the crosslinking area. Furthermore, mild or moderate staining of the BMZ was observed in 5 of the 16 specimens, and severe staining was observed in the other 11 specimens. Corneal staining, ranging from mild to severe, was observed in the crosslinking area of 16 of 16 specimens after CXL. The staining was observed in the subepithelial and stromal areas of the crosslinking area, without any endothelial involvement. The results
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