Miniature single-photon microscopes have been widely used to image neuronal assemblies in the brain of freely moving animals over the last decade. However, these systems have important limitations for imaging in-depth fine neuronal structures. We present a subcellular imaging single-photon device that uses an electrically tunable liquid crystal lens to enable a motion-free depth scan in the search of such structures. Our miniaturized microscope is compact ( 10    mm × 17    mm × 12    mm ) and lightweight ( ≈ 1.4    g ), with a fast acquisition rate (30–50 frames per second), high magnification ( 8.7 × ), and high resolution (1.4 μm) that allow imaging of calcium activity of fine neuronal processes in deep brain regions during a wide range of behavioral tasks of freely moving mice.

We present a motion-free system for microendoscopic imaging of biological tissues at variable focal depths. Fixed gradient index and electrically tunable liquid crystal lenses (TLCL) were used to build the imaging optical probe. The design of the TLCL enables polarization-independent and relatively low-voltage operation, significantly improving the energy efficiency of the system. A focal shift of approximately 74 ± 3 µm could be achieved by electrically controlling the TLCL using the driving frequency at a constant voltage. The potential of the system was tested by imaging neurons and spines in thick adult mouse brain sections and in vivo, in the adult mouse brain at different focal planes. Our results indicate that the developed system may enable depth-variable imaging of morpho-functional properties of neural circuitries in freely moving animals and can be used to investigate the functioning of these circuitries under normal and pathological conditions.

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