Dataset Information

Recording Neural Activity in Unrestrained Animals with Three-Dimensional Tracking Two-Photon Microscopy.

ABSTRACT: Optical recordings of neural activity in behaving animals can reveal the neural correlates of decision making, but brain motion, which often accompanies behavior, compromises these measurements. Two-photon point-scanning microscopy is especially sensitive to motion artifacts, and two-photon recording of activity has required rigid coupling between the brain and microscope. We developed a two-photon tracking microscope with extremely low-latency (360 ?s) feedback implemented in hardware. This microscope can maintain continuous focus on neurons moving with velocities of 3 mm/s and accelerations of 1 m/s2 both in-plane and axially. We recorded calcium dynamics of motor neurons and inter-neurons in unrestrained freely behaving fruit fly larvae, correlating neural activity with stimulus presentations and behavioral outputs, and we measured light-induced depolarization of a visual interneuron in a moving animal using a genetically encoded voltage indicator. Our technique can be extended to stabilize recordings in a variety of moving substrates.

SUBMITTER: Karagyozov D

PROVIDER: S-EPMC6287944 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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Recording Neural Activity in Unrestrained Animals with Three-Dimensional Tracking Two-Photon Microscopy.

Karagyozov Doycho D Mihovilovic Skanata Mirna M Lesar Amanda A Gershow Marc M

Cell reports 20181001 5

Optical recordings of neural activity in behaving animals can reveal the neural correlates of decision making, but brain motion, which often accompanies behavior, compromises these measurements. Two-photon point-scanning microscopy is especially sensitive to motion artifacts, and two-photon recording of activity has required rigid coupling between the brain and microscope. We developed a two-photon tracking microscope with extremely low-latency (360 μs) feedback implemented in hardware. This mic ...[more]

PMID: 30380425

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Project description:Background and objectiveTo develop a real-time recording system based on computer binocular vision and two-dimensional image feature extraction to accurately record mandibular movement in three dimensions.MethodsA computer-based binocular vision device with two digital cameras was used in conjunction with a fixed head retention bracket to track occlusal movement. Software was developed for extracting target spatial coordinates in real time based on two-dimensional image feature recognition. A plaster model of a subject's upper and lower dentition were made using conventional methods. A mandibular occlusal splint was made on the plaster model, and then the occlusal surface was removed. Temporal denture base resin was used to make a 3-cm handle extending outside the mouth connecting the anterior labial surface of the occlusal splint with a detection target with intersecting lines designed for spatial coordinate extraction. The subject's head was firmly fixed in place, and the occlusal splint was fully seated on the mandibular dentition. The subject was then asked to make various mouth movements while the mandibular movement target locus point set was recorded. Comparisons between the coordinate values and the actual values of the 30 intersections on the detection target were then analyzed using paired t-tests.ResultsThe three-dimensional trajectory curve shapes of the mandibular movements were consistent with the respective subject movements. Mean XYZ coordinate values and paired t-test results were as follows: X axis: -0.0037 ± 0.02953, P = 0.502; Y axis: 0.0037 ± 0.05242, P = 0.704; and Z axis: 0.0007 ± 0.06040, P = 0.952. The t-test result showed that the coordinate values of the 30 cross points were considered statistically no significant. (P<0.05).ConclusionsUse of a real-time recording system of three-dimensional mandibular movement based on computer binocular vision and two-dimensional image feature recognition technology produced a recording accuracy of approximately ± 0.1 mm, and is therefore suitable for clinical application. Certainly, further research is necessary to confirm the clinical applications of the method.

Project description:ObjectiveDevelopments in digital technology have permitted researchers to study mandibular movements. Here, the accuracy of a real-time, computerized, binocular, three-dimensional (3D) trajectory-tracking device for recording functional mandibular movements was evaluated.MethodsAn occlusal splint without the occlusal region was created based on a plaster cast of the lower dentition. The splint was rigidly connected with a target on its labial side and seated on the cast. The cast was then rigidly attached to the stage of a high-precision triaxial electronic translator, which was used to move the target-cast-stage complex. Half-circular movements (5.00-mm radius) in three planes (XOY, XOZ, YOZ) and linear movements along the x-axis were performed at 5.00 mm/s. All trajectory points were recorded with the binocular 3D trajectory-tracking device and fitted to arcs or lines, respectively, with the Imageware software. To analyze the accuracy of the trajectory-tracking device, the mean distances between the trajectory points and the fitted arcs or lines were measured, and the mean differences between the lengths of the fitted arcs' radii and a set value (5.00 mm) were then calculated. A one-way analysis of variance was used to evaluate the spatial consistency of the recording accuracy in three different planes.ResultsThe mean distances between the trajectory points and fitted arcs or lines were 0.076 ± 0.033 mm or 0.089 ± 0.014 mm. The mean difference between the lengths of the fitted arcs' radii and the set value (5.00 mm) was 0.025 ± 0.071 mm. A one-way ANOVA showed that the recording errors in three different planes were not statistically significant.ConclusionThese results suggest that the device can record certain movements at 5.00 mm/s, which is similar to the speed of functional mandibular movements. In addition, the recordings had an error of <0.1 mm and good spatial consistency. Thus, the device meets some of the requirements necessary for recording human mandibular movements.

Dataset Information

Recording Neural Activity in Unrestrained Animals with Three-Dimensional Tracking Two-Photon Microscopy.

Publications

Recording Neural Activity in Unrestrained Animals with Three-Dimensional Tracking Two-Photon Microscopy.

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