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Skin suturing and cortical surface viral infusion improves imaging of neuronal ensemble activity with head-mounted miniature microscopes. | LitMetric

Skin suturing and cortical surface viral infusion improves imaging of neuronal ensemble activity with head-mounted miniature microscopes.

J Neurosci Methods

Unit on Neural Circuits and Adaptive Behaviors, Clinical and Translational Neuroscience Branch, National Institute of Mental Health, Bethesda, MD, USA. Electronic address:

Published: November 2017

AI Article Synopsis

  • In vivo optical imaging of neural activity provides insights into brain functions, but traditional methods can cause tissue damage and limit clarity during imaging.
  • The researchers developed a new method using a wide-diameter glass pipette to infuse a viral calcium reporter while suturing the scalp to maintain optical window clarity during recovery.
  • This technique allowed efficient labeling of neurons, improved success rates, and enabled the recording of several hundred neurons in freely moving mice, with potential applications in other optical imaging methods.

Article Abstract

Background: In vivo optical imaging of neural activity provides important insights into brain functions at the single-cell level. Cranial windows and virally delivered calcium indicators are commonly used for imaging cortical activity through two-photon microscopes in head-fixed animals. Recently, head-mounted one-photon microscopes have been developed for freely behaving animals. However, minimizing tissue damage from the virus injection procedure and maintaining window clarity for imaging can be technically challenging.

New Method: We used a wide-diameter glass pipette at the cortical surface for infusing the viral calcium reporter AAV-GCaMP6 into the cortex. After infusion, the scalp skin over the implanted optical window was sutured to facilitate postoperative recovery. The sutured scalp was removed approximately two weeks later and a miniature microscope was attached above the window to image neuronal activity in freely moving mice.

Results: We found that cortical surface virus infusion efficiently labeled neurons in superficial layers, and scalp skin suturing helped to maintain the long-term clarity of optical windows. As a result, several hundred neurons could be recorded in freely moving animals.

Comparison With Existing Methods: Compared to intracortical virus injection and open-scalp postoperative recovery, our methods minimized tissue damage and dura overgrowth underneath the optical window, and significantly increased the experimental success rate and the yield of identified neurons.

Conclusion: Our improved cranial surgery technique allows for high-yield calcium imaging of cortical neurons with head-mounted microscopes in freely behaving animals. This technique may be beneficial for other optical applications such as two-photon microscopy, multi-site imaging, and optogenetic modulation.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5650079PMC
http://dx.doi.org/10.1016/j.jneumeth.2017.08.016DOI Listing

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