MicroRNAs　(miRNAs)　are　pivotal　in　regulating　biological　processes　such　as　cell　proliferation　and　disease　progression.　Traditional　miRNA　detection　methods　like　qRT-PCR　and　Northern　blotting　do　not　allow　for　monitoring　dynamic　changes　in　living　cells　and　typically　require　invasive　sample　collection.　This　research　presents　a　robust,　non-enzymatic　technique　known　as　the　Localized　Hybridization　Chain　Reaction　Amplifier　(LHCRA),　designed　for　real-time,　in　vivo　miRNA　imaging.　This　method　addresses　these　limitations　by　providing　rapid　and　sensitive　detection　of　miRNAs,　crucial　for　progressing　clinical　diagnostics　and　research.　The　LHCRA　utilizes　hairpin　chain　reaction　probes　embedded　within　self-assembled　DNA　micelles　(DM),　significantly　amplifying　miRNA　signals.　LHCRA　demonstrated　exceptional　performance,　with　a　detection　limit　of　100　pM　and　a　response　time　of　10　min.　Importantly,　it　effectively　differentiated　between　cancerous　and　normal　cells　using　clinical　peripheral　blood　samples,　showcasing　high　specificity　and　stability　under　physiological　conditions.　Additionally,　LHCRA　maintained　consistent　performance　in　complex　biological　media,　including　10　%　serum　and　2　U/mL　DNase　I,　confirming　its　broad　applicability　in　various　diagnostic　scenarios.　This　performance　highlights　LHCRA＇s　potential　as　a　transformative　tool　in　miRNA-based　diagnostics.　The　introduction　of　LHCRA　marks　a　significant　advancement　in　miRNA　detection　technology.　By　enabling　accurate,　non-invasive,　and　real-time　monitoring　of　miRNA　dynamics　within　living　cells,　this　method　offers　substantial　potential　to　enhance　diagnostic　accuracy　and　deepen　our　understanding　of　disease　mechanisms,　particularly　in　cancer.　Thus,　it　could　greatly　improve　therapeutic　strategies　and　patient　outcomes　in　clinical　environment.　©　2024