Imaging Large-Scale Neural Activity with Cellular Resolution in Awake, Mobile Mice
We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with i...
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          | Published in | Neuron (Cambridge, Mass.) Vol. 56; no. 1; pp. 43 - 57 | 
|---|---|
| Main Authors | , , , , | 
| Format | Journal Article | 
| Language | English | 
| Published | 
        United States
          Elsevier Inc
    
        04.10.2007
     Elsevier Limited  | 
| Subjects | |
| Online Access | Get full text | 
| ISSN | 0896-6273 1097-4199 1097-4199  | 
| DOI | 10.1016/j.neuron.2007.08.003 | 
Cover
| Abstract | We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to ∼2-5 μm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%. | 
    
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| AbstractList | We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to approximately 2-5 microm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%.We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to approximately 2-5 microm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%. We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to ∼2-5 μm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%. We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to 2-5 is a subset of m. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%. We report a technique for two-photon fluorescence imaging with cellular resolution in awake behaving mice with minimal motion artifact. The apparatus combines an upright table mounted two-photon microscope with a spherical treadmill consisting of a large air supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball’s upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their head remains motionless. Image sequences demonstrate that running-associated brain motion is limited to ~2–5 µm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom designed Hidden Markov Model based motion correction algorithm useful for post-processing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion induced false positive error rate of <5%. We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to approximately 2-5 microm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%. We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to ∼2-5 μm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%.  | 
    
| Author | Dombeck, Daniel A. Tank, David W. Adelman, Thomas L. Khabbaz, Anton N. Collman, Forrest  | 
    
| Author_xml | – sequence: 1 givenname: Daniel A. surname: Dombeck fullname: Dombeck, Daniel A. organization: Department of Molecular Biology, Carl Icahn Labs, Princeton University, Princeton, NJ 08544, USA – sequence: 2 givenname: Anton N. surname: Khabbaz fullname: Khabbaz, Anton N. organization: Department of Molecular Biology, Carl Icahn Labs, Princeton University, Princeton, NJ 08544, USA – sequence: 3 givenname: Forrest surname: Collman fullname: Collman, Forrest organization: Department of Molecular Biology, Carl Icahn Labs, Princeton University, Princeton, NJ 08544, USA – sequence: 4 givenname: Thomas L. surname: Adelman fullname: Adelman, Thomas L. organization: Department of Molecular Biology, Carl Icahn Labs, Princeton University, Princeton, NJ 08544, USA – sequence: 5 givenname: David W. surname: Tank fullname: Tank, David W. email: dwtank@princeton.edu organization: Department of Molecular Biology, Carl Icahn Labs, Princeton University, Princeton, NJ 08544, USA  | 
    
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/17920014$$D View this record in MEDLINE/PubMed | 
    
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| SubjectTerms | Algorithms Animals Astrocytes - physiology Behavior, Animal Brain - cytology Diagnostic Imaging - instrumentation Diagnostic Imaging - methods Equipment Design Exercise Test - methods Female Fitness equipment Luminescent Proteins - metabolism Male Markov Chains Mice Mice, Transgenic Models, Neurological Motor Activity - physiology Neurons Neurons - physiology Organic Chemicals - metabolism SYSNEURO Wakefulness - physiology Walking  | 
    
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| Title | Imaging Large-Scale Neural Activity with Cellular Resolution in Awake, Mobile Mice | 
    
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