AbstractCyclen‐peptide bioconjugates are usually prepared in multiple steps that require individual preparation and purification of the cyclic peptide and hydrophilic cyclen derivatives. An efficient strategy is discovered for peptide cyclization and functionalization toward lanthanide probe via three components intermolecular crosslinking on solid‐phase peptide synthesis with high conversion yield. Multifunctionality can be conferred by introducing different modular parts or/and metal ions on the cyclen‐embedded cyclopeptide. As a proof‐of‐concept, a luminescent Eu³⁺ complex and a Gd³⁺‐based contrasting agent for in vitro optical imaging and in vivo magnetic resonance imaging, respectively, are demonstrated through utilizing this preparation of cyclen‐embedded cyclic arginylglycylaspartic acid (RGD) peptide.