Fig. 1.
System and implementation of simultaneous ultrasound modulation and two-photon calcium imaging. (a) System design of the dual-modality system combining a two-photon laser scanning fluorescence microscope and a 30 MHz polymer ultrasound transducer. (b) The mechanical assembly including the head bar, O-ring, water container, and base support. (c) Representative two-photon calcium imaging of mouse visual cortex without and with ultrasound modulation. (d) Calcium transient traces for the neurons labeled in c. (e) Maximum-normalized calcium activity. (f) Statistics of ΔF/F0 before, during, and after ultrasound modulation for awake mice. (g) Statistics of ΔF/F0 before, during, and after ultrasound modulation for mice in anesthesia. (h) The recovering of the calcium activity in awake mice over time after the ultrasound was turned off. (i) Statistics of ΔF/F0 during the initial and the subsequent 90 s after the ultrasound was turned off in awake mice. **P < 0.01, *** or ###P < 0.001.