Miniature multiplexed label-free pH probe in vivo

Correlating in-brain pH fluctuations with the pathophysiology has been impeded by the lack of in vivo techniques to precisely determine local pH changes. Here, we developed an all-in-one pH probe for spatially-resolved and label-free pH sensing in vivo, based on a field-effect pH sensor, i.e., a light-addressable potentiometric sensor (LAPS), coupled to a flexible multimodal fiber. A readout photocurrent from the LAPS, elicited from a modulated light source, registers the localized surface potential change, proportional to the pH change. Upon simultaneous illuminations at multi-spot by a plurality of light sources with different modulation frequencies, pH changes at multiple designated spots are obtained via demultiplexing this photocurrent. To enable its in vivo applications, we combined the LAPS with a multimodal fiber fabricated by the convergence thermal drawing. Such fiber seamlessly integrates a multicore optical waveguide in the center for the light delivery, surrounded by electrodes for leading out photocurrent and serving as a pseudo-reference electrode, respectively. Such hybrid all-in-one pH probes can measure pH changes at 14 pixels simultaneously with a spatial resolution of 250 pm and a temporal resolution of 30 Hz. The pH sensitivity was characterized as 57.5 +/- 2.2 mV/pH homogeneously across all measurable pixels. Such probes have been implanted into the hippocampal formation of rats and their capabilities to capture pH changes at multiple pixels were evaluated at both physiological and pathological conditions. Technologies developed here represents a new class of in vivo chemical sensing technologies enabling the spatially-resolved investigation of intrinsic chemical signals in deep brain structures with high spatial and temporal resolutions.