Biohybrid Photonic Platform for Subcellular Stimulation and Readout of In Vitro Neurons

Targeted manipulation of neural activity via light has become an indispensable tool for gaining insights into the intricate processes governing single neurons and complex neural networks. To shed light onto the underlying interaction mechanisms, it is crucial to achieve precise control of individual neural activity, as well as a spatial read-out resolution on the nanoscale. Here, a versatile photonic platform with subcellular resolution for stimulation and monitoring of in-vitro neurons is demonstrated. Low-loss photonic waveguides are fabricated on glass substrates using nanoimprint lithography and featuring a loss of only -0.9 +/- 0.2 dB cm-1 at 489 nm and are combined with optical fiber-based waveguide-access and backside total internal reflection fluorescence microscopy. Neurons are grown on the bio-functionalized photonic chip surface and, expressing the light-sensitive ion channel Channelrhodopsin-2, are stimulated within the evanescent field penetration depth of 57 nm of the biocompatible waveguides. The versatility and cost-efficiency of the platform, along with the possible subcellular resolution, enable tailor-made investigations of neural interaction dynamics with defined spatial control and high throughput. Light-based manipulation of neural activity is vital for understanding single neurons and complex networks. Precise control and nanoscale spatial resolution are essential. A photonic platform using low-loss waveguides on glass substrates and optical fiber-based technology enables subcellular stimulation and monitoring of in vitro neurons. This versatile, cost-effective platform allows tailored studies of neural interaction dynamics with precise spatial control and high throughput.image