Imaging Brain Tissue with Quantum Light at Low Power.

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Light-induced tissue damage is a crucial limitation for traditional microscopy of the living brain, underscoring the need for new techniques that minimize exposure of samples to light. Here, we tested the hypothesis that quantum light, i.e., entangled photons, could detect brain structures at a lower excitation energy. In a proof of principle, we show microscopic images of fixed brain tissue in the hippocampus area created by fluorescence selective excitation in the process of entangled two-photon absorption in a scanning microscope. Quantum-enhanced entangled two-photon microscopy (TPM) had brain imaging capabilities at an unprecedented low excitation intensity of ∼3.6 × 107 photons/s, orders of magnitude lower than the excitation level for the classical two-photon fluorescence image obtained in the same microscope. The extremely low light probe intensity demonstrated in entangled TPM is of critical importance in the investigation of neural activity to minimize heating and photobleaching during repetitive imaging. It may have important functional implications in optogenetic technology, removing unintended heating and accumulated photodamage effects. This technology also opens avenues in spatially resolved brain tissue investigations with quantum light, providing new capabilities in local spectroscopy.