Abstract
<jats:title>ABSTRACT</jats:title> <jats:p> Hyperpolarised magnetic resonance (MR) enables real-time measurement of metabolic flux in living systems but remains difficult to deploy for cell-based studies because each hyperpolarisation event typically interrogates a single biological condition, limiting throughput, replication and longitudinal experimentation. Here we present a deployable microfluidic platform for parallel and longitudinal hyperpolarised <jats:sup>13</jats:sup> C metabolic phenotyping using standard MRI instrumentation. By combining microfluidics with spatially resolved MR spectroscopic imaging, the platform converts a single hyperpolarised preparation into multiple independent metabolic measurements without dedicated radiofrequency receive arrays or specialised instrumentation. We demonstrate reproducible discrimination of metabolically active and inactive samples, resolve cell-type-specific metabolic phenotypes, quantify biochemical and pharmacological perturbations, and recover metabolic exchange kinetics from parallel samples. Beyond increasing throughput, the platform enables repeated, non-destructive metabolic interrogation of the same recirculating three-dimensional cell cultures, allowing longitudinal phenotyping of living constructs rather than endpoint comparisons of independent samples. Across this study, 186 HP-MR measurements were acquired using only 44 polarisation events, corresponding to an approximately 4-fold increase in experimental throughput, while longitudinal monitoring reduced biological sample preparation 5-fold by following the same constructs over time. By lowering the technical barrier to hyperpolarised metabolic imaging while enabling both parallel and longitudinal metabolic phenotyping, this platform provides an accessible framework for drug discovery, microphysiological systems and patient-derived models. </jats:p>