Chemically micropatterned surfaces have broad applications in many fields. In this presentation, we report a new method for preparing chemically micropatterned surfaces by controlling and manipulating supported phospholipid monolayers as soft resist layers with molecular-level precision. First, we introduce self-assembled supported phospholipid monolayers on solid surfaces and use microcontact lift-up processes to create micropatterned phospholipid monolayers (with micrometer resolution) on the surface. Next, the micropatterned phospholipid monolayers can function as “soft” resist layers to protect underlying solid substrates and create either positive or negative chemically micropatterned surfaces during subsequent treatments. Unlike traditional “hard” resist layers which can only be removed by using harsh chemical treatments, this novel soft resist layer only comprises a single layer of compact phospholipid; therefore, it can be easily removed by water rinsing after the preparation of micropatterns. This method is also versatile. It can be applied to prepare protein microarray or silver patterns on solid substrates.