Dataset Information

Correction of z-motion artefacts to allow population imaging of synaptic activity in behaving mice.

ABSTRACT: KEY POINTS:Motion artefacts associated with motor behaviour are an inevitable problem of multiphoton imaging in awake behaving animals, particularly when imaging synapses. Correction of axial motion artefacts usually requires volumetric imaging resulting in slower rates of acquisition. We describe a method to correct z-motion artefacts that is easy to implement and allows population imaging of synaptic activity while scanning a single plane in a standard multiphoton microscope. The method uses a reference volume acquired in two colour channels - an activity reporter and an anatomical marker of blood vessels. The procedure estimates the z-displacement in every frame and applies an intensity correction in which the z intensity profile for each synapse is modelled as a Moffat function. We demonstrate that the method allows synaptic calcium signals to be collected from populations of synaptic boutons in mouse primary visual cortex during locomotion. ABSTRACT:Functional imaging of head-fixed, behaving mice using two-photon imaging of fluorescent activity reporters has become a powerful tool for studying the function of the brain. Motion artefacts are an inevitable problem during such experiments and are routinely corrected for in x and y dimensions. However, axial (z) shifts of several microns can also occur, leading to intensity fluctuations in structures such as synapses that are small compared to the axial point-spread function of the microscope. Here we present a simple strategy to correct z-motion artefacts arising over the course of a time-series experiment in a single optical plane. Displacement in z was calculated using dye-filled blood vessels as an anatomical marker, providing high contrast images and accuracy to within ?0.1 µm. The axial profiles of ROIs corresponding to synapses were described using a Moffat function and this 'ROI-spread function' used to correct activity traces on an ROI-by-ROI basis. We demonstrate the accuracy and utility of the procedures in simulation experiments using fluorescent beads and then apply them to correcting measurements of synaptic activity in populations of vasoactive-intestinal peptide (VIP) interneurons expressing the synaptic reporter SyGCaMP6f. Correction of z-motion artefacts had a substantial impact on the apparent correlation between synaptic activity and running speed, demonstrating the importance of correcting these when performing imaging experiments in awake mice.

SUBMITTER: Ryan TM

PROVIDER: S-EPMC7318612 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Correction of z-motion artefacts to allow population imaging of synaptic activity in behaving mice.

Ryan Thomas Michael TM Hinojosa Antonio Jesus AJ Vroman Rozan R Papasavvas Christoforos C Lagnado Leon L

The Journal of physiology 20200303 10

<h4>Key points</h4>Motion artefacts associated with motor behaviour are an inevitable problem of multiphoton imaging in awake behaving animals, particularly when imaging synapses. Correction of axial motion artefacts usually requires volumetric imaging resulting in slower rates of acquisition. We describe a method to correct z-motion artefacts that is easy to implement and allows population imaging of synaptic activity while scanning a single plane in a standard multiphoton microscope. The metho ...[more]

PMID: 32020615

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Project description:Motion artefacts (MAs) are induced within EEG data collected simultaneously with fMRI when the subject's head rotates relative to the magnetic field. The effects of these artefacts have generally been ameliorated by removing periods of data during which large artefact voltages appear in the EEG traces. However, even when combined with other standard post-processing methods, this strategy does not remove smaller MAs which can dominate the neuronal signals of interest. A number of methods are therefore being developed to characterise the MA by measuring reference signals and then using these in artefact correction. These methods generally assume that the head and EEG cap, plus any attached sensors, form a rigid body which can be characterised by a standard set of six motion parameters. Here we investigate the motion of the head/EEG cap system to provide a better understanding of MAs. We focus on the reference layer artefact subtraction (RLAS) approach, as this allows measurement of a separate reference signal for each electrode that is being used to measure brain activity. Through a series of experiments on phantoms and subjects, we find that movement of the EEG cap relative to the phantom and skin on the forehead is relatively small and that this non-rigid body movement does not appear to cause considerable discrepancy in artefacts between the scalp and reference signals. However, differences in the amplitude of these signals is observed which may be due to differences in geometry of the system from which the reference signals are measured compared with the brain signals. In addition, we find that there is non-rigid body movement of the skull and skin which produces an additional MA component for a head shake, which is not present for a head nod. This results in a large discrepancy in the amplitude and temporal profile of the MA measured on the scalp and reference layer, reducing the efficacy of MA correction based on the reference signals. Together our data suggest that the efficacy of the correction of MA using any reference-based system is likely to differ for different types of head movement with head shake being the hardest to correct. This provides new information to inform the development of hardware and post-processing methods for removing MAs from EEG data acquired simultaneously with fMRI data.

Dataset Information

Correction of z-motion artefacts to allow population imaging of synaptic activity in behaving mice.

Publications

Correction of z-motion artefacts to allow population imaging of synaptic activity in behaving mice.

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