Methods for the analysis of cell secretions at the single-cell level only provide semiquantitative endpoint readouts. Here we describe a microwell array for the real-time spatiotemporal monitoring of extracellular secretions from hundreds of single cells in parallel. The microwell array incorporates a gold substrate with arrays of nanometric holes functionalized with receptors for a specific analyte, and is illuminated with light spectrally overlapping with the device’s spectrum of extraordinary optical transmission. Spectral shifts in surface plasmon resonance resulting from analyte–receptor bindings around a secreting cell are recorded by a camera as variations in the intensity of the transmitted light while machine-learning-assisted cell tracking eliminates the influence of cell movements. We used the microwell array to characterize the antibody-secretion profiles of hybridoma cells and of a rare subset of antibody-secreting cells sorted from human donor peripheral blood mononuclear cells. High-throughput measurements of spatiotemporal secretory profiles at the single-cell level will aid the study of the physiological mechanisms governing protein secretion.

在单细胞水平上分析细胞分泌物的方法仅提供半定量的终点读数。在这里，我们描述了一种微孔阵列，用于并行实时时空监测数百个单细胞的细胞外分泌物。微孔阵列包含一个金基板，该基板上有纳米孔阵列，这些纳米孔阵列通过特定分析物的受体进行功能化，并用与设备的非凡光学传输光谱重叠的光谱进行照明。分泌细胞周围的分析物与受体结合引起的表面等离振子共振的光谱变化被相机记录为透射光强度的变化，而机器学习辅助的细胞跟踪消除了细胞运动的影响。我们使用微孔阵列来表征杂交瘤细胞和从人类供体外周血单核细胞中分选的罕见抗体分泌细胞子集的抗体分泌谱。单细胞水平的时空分泌谱的高通量测量将有助于研究控制蛋白质分泌的生理机制。