Fig. 4: The manipulation of fCOAT system on neural activities.

a Diagram illustrating the experimental scheme used to demonstrate the ability of fCOAT focusing to elicit stronger calcium signals through 200-μm-thick mouse brain tissue. The black dashed line indicates the positional relationship between the target neuron and the focus. b The merge images of the expression of photosensitive protein and calcium indicator of the target neuron. Scale bar, 10 μm. c Targeted activation of calcium signaling in a representative neuron after focusing through the brain slice. Green: jGCaMP6s fluorescence signals. Orange: excitation beam intensity through the brain slice with and without wavefront shaping. The white dotted line: the location of the optimized focus and the boundaries of individual neuron. d Quantitative analysis of jGCaMP6s fluorescence signals obtained from the neuron in c. Shadows are given by the upper and lower limits of stimulations. e The histogram representation from the data in d. Comparison of peak neural response (mean ± SE) to different types of stimuli, ****P < 0.0001, strong (73.76 ± 5.41% ΔF⁄F) versus weak (23.47 ± 1.89% ΔF⁄F). f Individual traces of jGCaMP6s fluorescence in different C1V1 neurons under 589 nm illumination using focus and speckle light fields. g The statistical results of neuron data in f and the corresponding analysis between different types of stimuli. Error bars:1.5 SE of five measurements of peak neural responses. h The results of the relationship between the focus PBR and the calcium signals enhancements. The shadow is formed by moving the trend line up and down. i Targeted activation of calcium signaling in representative neurons after focusing through the brain slices. Red: Cal590 fluorescence signals. Green: the expression of the used opsin ChR2. Blue: excitation beam intensity through the brain slice with or without wavefront shaping. The white dotted line: the boundaries of individual neurons. Scale bar, 20 μm. j Individual traces of Cal590 fluorescence in different ChR2 neurons under 488 nm illumination using extended focus and speckle light fields.