Neurotransmitter release relies on the regulated fusion of synaptic vesicles (SVs) that are tightly packed within the presynaptic bouton of neurons. The ability of SVs to dynamically recycle while remaining clustered at the presynapse is unknown but suggests anchoring mechanisms that regulate SV function. One potential anchor is a family of phosphoproteins that interact with presynaptic SVs, called synapsins. Synapsins tightly bind SVs and form biomolecular condensates via liquid-liquid phase separation which can potentially help trap SVs at the presynapse. An alternative model for synapsin’s clustering function is its ability to tetramerize, thereby forming a network of between SVs. Synapsin 2a (Syn2a) is the only isoform capable to rescuing the synaptic depletion induced by synapsin triple knockout. We investigate the role of Syn2a in SV clustering using super-resolution dual-pulse sub-diffractional tracking of internalised molecules and sptPALM to simultaneously track Syn2a-mEos3.2 molecules and single SVs from the reserve pool in live hippocampal neurons. Reserve SVs display lower presynaptic mobility compared to recycling SVs and both pools exhibit a mobile axonal pool. Triple knockout of Synapsin 1-3 genes increased the mobility of reserve SVs. Re-expression of wild-type Syn2a (Syn2aWT), but not the tetramerization-deficient mutant K337Q (Syn2aK337Q), fully rescued these effects. Single-particle tracking revealed that Syn2aK337QmEos3.2 exhibited altered activity-dependent presynaptic translocation and nanoclustering. Syn2a tetramerization therefore controls its own presynaptic nanoclustering and contributes to the dynamic immobilisation of the SV reserve pool.