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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
Injection of Viral Tracers by Nanoject
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bd8qi9vw BICCN, Allen Institute for Brain Science This protocol describes the delivery of a neuronal tracer using the Nanoject II. The surgery uses a stereotaxic system to target specific brain coordinates in the mouse. Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Injection of Viral Tracers by Nanoject. protocols.io https://dx.doi.org/10.17504/protocols.io.bd8qi9vw 2021-04-15 09:14:30
Mouse Cardiac Perfusion Fixation and Brain Collection
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.be2djga6 BICCN, Allen Institute for Brain Science This protocol describes the procedures for intracardiac perfusion fixation of postnatal mice, including anesthesia, exsanguination, fixation, brain removal and post-fixation storage. Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 4 2020 Allen Institute for Brain Science 2020. Mouse Cardiac Perfusion Fixation and Brain Collection. protocols.io https://dx.doi.org/10.17504/protocols.io.be2djga6 2021-04-15 09:14:26
Sequential smFISH
 
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ZengU19 BICCN Grant DOI:10.17504/protocols.io.y6nfzde BICCN, Allen Institute for Brain Science We have developed a multiplexed single molecule FISH protocol for use at the Institute. This protocol was optimized on human tissue, but will work on mouse tissue as well. It was adapted from Lyubimova et. al., Nature Protocols, 2013.  Allen Institute 3 2019 ZengU19 BICCN Grant 2019. Sequential smFISH. protocols.io https://dx.doi.org/10.17504/protocols.io.y6nfzde 2021-04-15 09:14:27
Isolation of Nuclei from Adult Human Brain Tissue for 10x Genomics Platform
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bq64mzgw BICCN, Allen Institute for Brain Science Isolation of nuclei from frozen adult human brain tissue or thawed and microdissected brain tissue sections for RNA-seq analysis. Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U01MH114812. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2021 Allen Institute for Brain Science 2021. Isolation of Nuclei from Adult Human Brain Tissue for 10x Genomics Platform. protocols.io https://dx.doi.org/10.17504/protocols.io.bq64mzgw 2021-04-15 09:14:26
FACS Single Cell Sorting
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.besbjean BICCN, Allen Institute for Brain Science This protocol provides step by step instructions for performing single-cell sorting by using BD FACSAria II machine or BD FACSAria Fusion machine and BD FACSDiva software.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 3 2020 Allen Institute for Brain Science 2020. FACS Single Cell Sorting. protocols.io https://dx.doi.org/10.17504/protocols.io.besbjean 2021-04-15 09:14:30
50 mM Phosphate Buffer (with 0.02% sodium azide option)
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bezjjf4n BICCN, Allen Institute for Brain Science 50 mM Phosphate Buffer can be used as a general buffer solution for multiple applications. It is suitable for use as a buffer for specimen embedding, as part of brain preparation associated with TissueCyte 2P Serial Imaging, described in PF0315 TissueCyte Specimen Embedding Acrylamide Coembedding.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. 50 mM Phosphate Buffer (with 0.02% sodium azide option). protocols.io https://dx.doi.org/10.17504/protocols.io.bezjjf4n 2021-04-15 09:14:30
Pulling Pipettes using the DMZ-Zeitz Universal
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bdpfi5jn BICCN, Allen Institute for Brain Science Protocol is used for the manufacture of glass pipettes for patch clamp electrophysiology using the DMZ-Zeitz Universal puller.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 2 2020 Allen Institute for Brain Science 2020. Pulling Pipettes using the DMZ-Zeitz Universal. protocols.io https://dx.doi.org/10.17504/protocols.io.bdpfi5jn 2021-04-15 09:14:33
Degassing Reagents
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bg4wjyxe BICCN, Allen Institute for Brain Science This protocol is used for the degassing of critical reagents used in both the colorimetric and fluorescent in situ hybridization (ISH). This is crucial in eliminating hybridization bubbles from forming on the tissue. Hybridization bubbles result in no expression where they are located on the tissue.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Degassing Reagents. protocols.io https://dx.doi.org/10.17504/protocols.io.bg4wjyxe 2021-04-15 09:14:34
PBS with 0.1% Sodium Azide
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bfzsjp6e BICCN, Allen Institute for Brain Science This protocol describes the method for preparing PBS with 0.1% Sodium Azide for storage and/or processing of fixed tissue.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. PBS with 0.1% Sodium Azide. protocols.io https://dx.doi.org/10.17504/protocols.io.bfzsjp6e 2021-04-15 09:14:33
Subcutaneous Injection Adult Mouse
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bezdjf26 BICCN, Allen Institute for Brain Science This protocol describes general procedures used for subcutaneous injection in adult mice.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Subcutaneous Injection Adult Mouse. protocols.io https://dx.doi.org/10.17504/protocols.io.bezdjf26 2021-04-15 09:14:31
PCR Prep from Glycerol Stock for IVT Reactions
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.brt2m6qe BICCN, Allen Institute for Brain Science This protocol is used for the PCR reaction from clone templates in glycerol stocks generating transcript templates ready for IVT reaction.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number 1U19MH114830-01. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health Allen Institute 1 2021 Allen Institute for Brain Science 2021. PCR Prep from Glycerol Stock for IVT Reactions. protocols.io https://dx.doi.org/10.17504/protocols.io.brt2m6qe 2021-04-15 09:14:52
Sequencing open chromatin of single cell nuclei: snATAC-seq
 
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Sebastian Preissl, Xinxin Wang, Bing Ren DOI:10.17504/protocols.io.pjudknw BICCN Protocol for applying the TN5-based ATAC-seq method to the analysis of the chromatin of single cell nuclei isolated from brain tissue, as performed within the Center for Epigenomics of the Mouse Bain Atlas (CEMBA) and in coordination with the BRAIN Initiative Cell Census Network (BICCN). Cellular and Molecular Medicine, University of California, San Diego, Cellular and Molecular Medicine, University of California, San Diego, Ludwig Institute and Cellular and Molecular Medicine, University of California, San Diego 1 2018 Sebastian Preissl, Xinxin Wang, Bing Ren 2018. Sequencing open chromatin of single cell nuclei: snATAC-seq. protocols.io https://dx.doi.org/10.17504/protocols.io.pjudknw 2021-04-15 09:14:49
Core Protocol for Serial Imaging of Fluorescently-Labeled Mouse Brain on the TissueCyte 1000 System
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bdvmi646 BICCN, Allen Institute for Brain Science This protocol describes the instructions for setup and scanning of fluorescently-labeled tissue sections mounted in agarose and attached to a slide using the TissueCyte 1000 block face 2-photon system.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 2 2020 Allen Institute for Brain Science 2020. Core Protocol for Serial Imaging of Fluorescently-Labeled Mouse Brain on the TissueCyte 1000 System. protocols.io https://dx.doi.org/10.17504/protocols.io.bdvmi646 2021-04-15 09:14:40
cDNA Synthesis
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.9geh3te BICCN, Allen Institute for Brain Science This protocol describes the synthesis of first-strand cDNA from total RNA using the Invitrogen SuperScript III Reverse Transcriptase system. Allen Institute 1 2019 Allen Institute for Brain Science 2019. cDNA Synthesis. protocols.io https://dx.doi.org/10.17504/protocols.io.9geh3te 2021-04-15 09:14:54
Electrode Internal Solution with Biocytin II
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bepnjdme BICCN, Allen Institute for Brain Science This SOP provides instructions to prepare Internal Solution II used for simultaneous patch clamp electrophysiology experiments to probe circuit connectivity.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Electrode Internal Solution with Biocytin II. protocols.io https://dx.doi.org/10.17504/protocols.io.bepnjdme 2021-04-15 09:14:37
Mouse Whole Cell Tissue Processing for 10x Genomics Platform
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bsfdnbi6 BICCN, Allen Institute for Brain Science This protocol describes how to isolate tissue from various ROI’s of the mouse brain followed by preparation of a highly concentrated single cell suspension for transcriptomic profiling on 10x Genomics platform.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 8 2021 Allen Institute for Brain Science 2021. Mouse Whole Cell Tissue Processing for 10x Genomics Platform. protocols.io https://dx.doi.org/10.17504/protocols.io.bsfdnbi6 2021-04-15 09:14:58
Preparation of Gelatin Slide Coating Solution
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bfzvjp66 BICCN, Allen Institute for Brain Science This protocol describes the preparation of gelatin solution to be used for coating glass microscope slides.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Preparation of Gelatin Slide Coating Solution. protocols.io https://dx.doi.org/10.17504/protocols.io.bfzvjp66 2021-04-15 09:14:41
Artificial Cerebrospinal Fluid IV (ACSF.IV)
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bepsjdne BICCN, Allen Institute for Brain Science This protocol describes making Artificial Cerebrospinal Fluid IV (ACSF.IV). ACSF.IV is used for multiple applications including incubation of fresh mouse brain slices prior to electrophysiological recording.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Artificial Cerebrospinal Fluid IV (ACSF.IV). protocols.io https://dx.doi.org/10.17504/protocols.io.bepsjdne 2021-04-15 09:14:43
Maintenance of TissueCyte System
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.ber3jd8n BICCN, Allen Institute for Brain Science This protocols provides detailed instructions for procedures needed for maintenance of the TissueCyte scanning systems.Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Maintenance of TissueCyte System. protocols.io https://dx.doi.org/10.17504/protocols.io.ber3jd8n 2021-04-15 09:14:45
3D Reconstruction of Neurons in Vaa3D
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bcseiwbe BICCN, Allen Institute for Brain Science This protocol is used to generate accurate digital representations of neuron morphologies from a variety of brain regions and species. Each reconstruction captures the positions and thicknesses of the soma, dendrites and axon of a biocytin-filled cell within a slice of brain tissue. To generate the reconstruction we use an image stack containing ~200-700 serial 2D images that captures the full extent of the cell within the slice. We use the Vaa3D (Terafly) program with a Kazom’s Mozak user interface (Mozak for short) to visualize the 2D images in 3D. Once the stack is loaded in Mozak, our reconstruction is generated by placing nodes in 3D space. The placement of these nodes is dependent on the signal in the images. Our final output, an SWC text file (.swc format), contains many thousand rows. Each row contains a node ID, an x, y, z coordinate, radius value, neurite type, and parent node ID. After tracing is complete, we preform post processing to provide radius values, check for errors and consistency then the SWCs are uploaded into our Laboratory Information Management System. Allen Institute 1 2020 Allen Institute for Brain Science 2020. 3D Reconstruction of Neurons in Vaa3D. protocols.io https://dx.doi.org/10.17504/protocols.io.bcseiwbe 2021-04-15 09:15:06

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