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. 2022 Mar 7;12:3678. doi: 10.1038/s41598-022-07313-1

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Characterization of ASAP3-mCyRFP3 (ASAP3-R3). (a) Representative images of ASAP3 fusions to various RFPs expressed in primary rat hippocampal neurons acquired on a spinning-disk confocal microscope using Improvision Volocity 6.0 and processed in NIH Fiji 2.1. (b) Laser-scanning confocal images of ASAP3-R3 in a rat hippocampal neuron. Images were acquired with Zeiss Zen Blue software and processed in Fiji 2.1. (c) Above, normalized excitation spectra of mCyRFP3 and mEGFP, showing a single excitation wavelength can be used. Below, normalized emission spectra of mCyRFP3 and mEGFP, showing their emissions are separable (d) Steady-state voltage responsivity of ASAP3-R3. Error bars represent standard error of the mean (n = 8 HEK293 cells). Graphs were made in Microsoft Excel for Mac 16.