There has been extensive interest in the micro and nanoscale manipulation of various substrates in the past few decades. One promising technique is dip-pen nanolithography which has shown the capability to pattern substrates of all forms including, tissue-derived substrates. Patterning of tissue-derived substrates is of particular interest, as it would facilitate studies into controlling cell morphology and cell-substrate interaction. To expand the field into this area both peptides and bioactive collagen-binding peptide-linked biomolecules were patterned to the inner collagenous zone of the Bruch's membrane (BM). Collagen-binding peptide, and extra cellular matrix (ECM) proteins laminin and fibronectin were patterned on the BM and SiO(x). The lithographic protocol was facilitated by Triton X-100 which was used to clean the tissue-derived construct after harvesting. This produced a collagen-exposed BM which was more hydrophilic (contact angle 67 degrees +/-8.49 degrees) surface compared with other cleaning methods but it maintained similar surface roughness (root-mean-square) 80+/-18 nm and collagen exposure. This type of surface can be readily patterned with the chosen inks under lower humidity conditions.