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. 2018 Nov 2;8:16262. doi: 10.1038/s41598-018-34472-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Optical setup. (A) A custom-made microscope comprising one illumination path (green) and two detection paths (red) is coupled to the animal brain using an image guide attached to a micro-objective. The central element of the fiberscope setup is a DMD, which is used to create a matrix of illumination and detection pinholes. The optical setup is fully described in the methods. Lenses L₃ and L₄ are off-axis to partly straighten up the multipoint-scanning confocal image at the camera. DBS: dichroic beam splitter. M: mirror. BS(90:10): beam splitter with 90% reflection and 10% transmission. F: filter. (B) Picture of an unrestrained (freely behaving) mouse in its cage with the fiberscope probe fixed on the skull. (C) Fixation of the image guide (ferrule shown in yellow) and GRIN lens (ferrule shown in cyan) to the skull. The image guide is attached to the GRIN lens using a connection device (piece 1). The GRIN lens is fixed to the skull using a head plate (piece 2) that is positioned using a micromanipulator.