Viral Injection and Cranial Window Implantation for In Vivo Two-Photon Imaging

  • Gordon B. Smith
  • David Fitzpatrick
Part of the Methods in Molecular Biology book series (MIMB, volume 1474)


Two-photon (2P) imaging has proven to be a powerful tool for investigating neural structure and function both in brain slices and in intact systems. In vivo 2P imaging presents significant challenges in sample preparation, which are exacerbated in non-murine species. Here, we describe procedures for the effective virally mediated labeling of neurons and for the implantation of cranial windows for imaging. The procedures described here are applicable to a range of species, including mice, and are routinely used in ferrets and tree shrews to provide large-scale labeling of cortical volumes and high-quality imaging data.

Key words

Two-photon In vivo Microscopy Imaging Viral expression Genetically encoded calcium sensor 



We would like to thank Dominique Ouimet for helpful discussions on surgical procedures, Yishai Elyada for the headplate design and discussions on cranial window implants, and Daniel Wilson for images of dendritic spines and for comments on this manuscript.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Functional Architecture and Development of Cerebral CortexMax Planck Florida Institute for NeuroscienceJupiterUSA

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