The evaluation of the surgical margins is a major concern in surgical pathology, and marking of surgical margins with substances such as alcian blue, Tipp-ex, artist's pigments, colored gelatin, starch, erythrocyte layers, etc. was recommended for this purpose; Indian ink and tissue marking dyes are widely used. As there is no systematic study comparing tissue marking dyes and Indian ink as the most common substances used for the purpose, this study was conducted to compare the two. Penetration into the tissue, brightness under the microscope, the spreading area of one drop of dye on tissue paper, the intensity of colors, and unit price were compared for each of the five colors of Rotring's Indian ink and Thermo-Shandon's tissue marking dyes, applied on reduction mammoplasty specimens. Rotring's Indian ink is proved to be just as effective as Thermo-Shandon's tissue marking dye and bares the majority of the characteristics of a perfect staining substance, which are easily applied, quickly fixed, durable and cheap, contain no potential contaminants, be work safe, would not smudge/stain surrounding tissues, and look bright under the microscope without obscuring the view.