Journal: The Journal of Neuroscience

Article Title: Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

doi: 10.1523/JNEUROSCI.1736-22.2022

Figure Lengend Snippet: TIPM produces reliable, directional, and magnitude-dependent responses following roll tilts. A , A 4 dpf larval zebrafish mounted in agarose for roll stimuli on a mirror galvanometer. B , Schematic of our experimental paradigm. Baseline fluorescence (used for normalization) is measured when the platform is horizontal. The galvanometer is then stepped and held at an eccentric angle (Stimulus) where fluorescence is not recorded, then quickly returned to horizontal, whereupon fluorescent recording begins (Response). C , Voltage trace from galvanometer during a 10, 20, and 30° step to the left. D , Feedback voltage from the galvanometer during a step to 30°. E , Slices from a two-photon volume of Tg(nefma:GAL4);Tg(UAS:GCaMP6s) fish. Dashed yellow overlays indicate pixels that correspond to analyzed vestibulospinal neurons. Yellow square shows close-up of a single analyzed cell. Scale bar, 20 µm. F , Normalized fluorescence traces for all trials of one neuron during baseline and response to an ipsilateral 30° roll. G , Distribution of coefficients of variation of peak ΔF/F values across 30° step trials for responsive neurons ( n = 69 neurons). H , Mean normalized fluorescence traces for one neuron during baseline and response to ipsilateral and contralateral roll steps of varying magnitudes (10, 20, 30°). I , Mean peak ΔF/F responses across all responsive neurons for ipsilateral and contralateral rolls of 10, 20, and 30° magnitudes. Error bars indicate ± SEM. J , Distribution of directionality indices (see above, Materials and Methods) across all responsive neurons.

Article Snippet: Galvanometer control was done in MATLAB 2019b software (MathWorks) using the Data Acquisition Toolbox to interface with a data acquisition card (PCIe-6363, National Instruments).

Techniques: Fluorescence

Journal: The Journal of Neuroscience

Article Title: Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

doi: 10.1523/JNEUROSCI.1736-22.2022

Figure Lengend Snippet: Variability in responses arises predominantly from intrinsic sources. A , Timeline of an experiment of repeated imaging of the same fish across two mounts on the galvanometer. B , Two-photon volumes of vestibulospinal neurons in a Tg(nefma:GAL4);Tg(UAS:GCaMP6s) larva during two sequential mounting and imaging experiments. Colored overlays indicate the same neurons located in two separate volumes taken across the two mounts. C , Mean fluorescence traces (± SD) during the baseline period and following an ipsilateral roll during the first and second mount for two neurons (orange and blue traces correspond to colored neuron overlays). D , Mean peak response during trials in the first mount experiment are strongly correlated with the mean peak response from the second mount (ρ = 0.83). E , Peak response on a single experimental trial is strongly correlated with peak response on the subsequent trial within the same experiment (ρ = 0.79).

Article Snippet: Galvanometer control was done in MATLAB 2019b software (MathWorks) using the Data Acquisition Toolbox to interface with a data acquisition card (PCIe-6363, National Instruments).

Techniques: Imaging, Fluorescence

Journal: The Journal of Neuroscience

Article Title: Tilt in Place Microscopy: a Simple, Low-Cost Solution to Image Neural Responses to Body Rotations

doi: 10.1523/JNEUROSCI.1736-22.2022

Figure Lengend Snippet: Ventral vestibulospinal neurons respond to impulse stimuli in a nondirectional, magnitude-independent manner. A , Voltage trace corresponding to feedback from galvanometer during a 10, 15, and 30° impulse step to the left. B , Feedback trace during the impulse step to 30°. C , Normalized fluorescence traces for all trials of one neuron during baseline and response to an ipsilateral 30° impulse step. Note the lower scale on the vertical axis relative to – . D , Distribution of coefficients of variation of peak fluorescent intensity across 30° step trials for responsive neurons ( n = 22 neurons). E , Mean normalized fluorescence traces for one neuron during baseline and response to an ipsilateral and contralateral impulse step of varying magnitudes (10, 20, 30°). F , Mean peak ΔF/F responses across all responsive neurons for ipsilateral and contralateral impulse steps of 10, 15, and 30° magnitudes. Error bars indicate ± SEM. G , Distribution of directionality indices (see above, Materials and Methods) across all responsive neurons. H , Spatial location of impulse-responsive (green) or nonresponsive (gray) vestibulospinal somata relative to the Mauthner neuron lateral dendrite in microns. I , Distribution of dorsoventral position of vestibulospinal neuron bodies relative to the Mauthner lateral dendrite in microns for impulse-responsive (green) and nonresponsive (gray) neurons. Solid lines represent the distribution of all responsive or nonresponsive neurons across all fish. Error bars represent ± 1 bootstrapped SD.

Article Snippet: Galvanometer control was done in MATLAB 2019b software (MathWorks) using the Data Acquisition Toolbox to interface with a data acquisition card (PCIe-6363, National Instruments).

Techniques: Fluorescence