In this paper, two kinds of stamps (squares (a×a)) separated by spacing b, the values of a and b were varied from 2.5 µm to 50 µm), i.e. positive and negative stamps, were prepared. The stamps inked with the rhodamine-labeled bovine serum albumin (BSA) were then microcontacted with the aldehyde-functionalized titanium surfaces. Water contact angle and X-ray photoelectron spectrum (XPS) indicated that BSA can be covalently immobilized on aldehyde modified titanium surface by microcontact printing. The experimental results of CLSM showed that the patterns with resolution from 2.5 µm to 50 µm were obtained successfully. Both positive stamp and negative stamp were deformed when the value of a was less than or equal to 5 µm, which resulted in replication errors. Furthermore, the larger spacing (50 µm) resulted in stamp collapse when the value a of the positive stamp was less than or equal to 10 µm, leading to whole fluorescence on substrates.
