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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
50 mM Phosphate Buffer (with 0.02% sodium azide option)
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.bezjjf4n 2021-03-29 03:12:47
cDNA Synthesis
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.besejebe 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.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. cDNA Synthesis. protocols.io dx.doi.org/10.17504/protocols.io.besejebe 2021-03-29 03:12:50
Tissue Quality Evaluation for Brain Perfusion/Dissection Specimens
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bd8yi9xw BICCN, Allen Institute for Brain Science This protocol provides guidance in the assessment of quality of tissues post perfusion and dissection and applies to all such procedures. Perfusion and dissection quality will be categorized on the following scale: perfect, good, fair, and poor. This SOP also presents written descriptions, and images, of specific category attributes. 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. Tissue Quality Evaluation for Brain Perfusion/Dissection Specimens. protocols.io dx.doi.org/10.17504/protocols.io.bd8yi9xw 2021-03-29 03:14:29
Patch-Seq Internal Solution with Biocytin
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.besdjea6 BICCN, Allen Institute for Brain Science This SOP provides instruction to prepare Internal Solution which is used for patch clamp electrophysiology, modified for cleanliness and mRNA capture and sustainability.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. Patch-Seq Internal Solution with Biocytin. protocols.io dx.doi.org/10.17504/protocols.io.besdjea6 2021-03-29 03:14:30
Bright Field Image Capture with Zeiss AxioImager
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bdpmi5k6 BICCN, Allen Institute for Brain Science This protocol describes the system that is used to capture Z stack images which are reassembled post-processing into 3D reconstructions.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. Bright Field Image Capture with Zeiss AxioImager. protocols.io dx.doi.org/10.17504/protocols.io.bdpmi5k6 2021-03-29 03:14:34
Slice Preparation and Blockface Imaging for Electrophysiology - Mouse
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bdpqi5mw BICCN, Allen Institute for Brain Science This protocol describes the procedure for transcardial perfusion of an adult mouse, followed by dissection, embedding, slicing, and blockface imaging of the mouse brain using either a Leica vibratome or a compresstome. The output tissue is suitable for use in electrophysiological recordings and/or live cell filling applications.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. Slice Preparation and Blockface Imaging for Electrophysiology - Mouse. protocols.io dx.doi.org/10.17504/protocols.io.bdpqi5mw 2021-03-29 03:14:41
Slice Preparation with Tissue Dissociation - Mouse Protocol
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bq6wmzfe BICCN, Allen Institute for Brain Science This protocol describes the procedure for transcardial perfusion of an adult mouse, followed by brain dissection, embedding, slicing, blockface imaging, ROI microdissection, and tissue dissociation. The output of this protocol is a cell suspension that can be sorted via FACS for single-cell transcriptional profiling.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 2021 Allen Institute for Brain Science 2021. Slice Preparation with Tissue Dissociation - Mouse Protocol. protocols.io dx.doi.org/10.17504/protocols.io.bq6wmzfe 2021-03-29 03:14:47
Human Neurosurgical Tissue Processing
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bet4jeqw BICCN, Allen Institute for Brain Science This protocol describes the procedure for receiving human surgical tissue, followed by trimming, mounting, embedding, and slicing under sterile conditions in order to produce tissue suitable for both slice culture as well as acute slice electrophysiology 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 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 5 2020 Allen Institute for Brain Science 2020. Human Neurosurgical Tissue Processing. protocols.io dx.doi.org/10.17504/protocols.io.bet4jeqw 2021-03-29 03:15:00
TissueCyte Installation And Alignment Guide
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bdpti5nn BICCN, Allen Institute for Brain Science This protocol outlines some of the steps that are required to align a TissueCyte 1000 serial two-photon imaging system and install commonly replaced parts.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. TissueCyte Installation And Alignment Guide. protocols.io dx.doi.org/10.17504/protocols.io.bdpti5nn 2021-03-29 03:15:00
PBS with 0.1% Sodium Azide
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.bfzsjp6e 2021-03-29 03:12:57
DAB Solutions
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bew7jfhn BICCN, Allen Institute for Brain Science 3, 3´ Diaminobenzidine (DAB) is a commonly used chromogen for histological staining. In the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol and xylene. Peroxidase catalyzes hydrogen peroxide oxidation of substrate DAB. This SOP provides instructions to prepare four different formulations of DAB. DAB for Immunohistochemical staining (IHC), DAB for Acetylcholinesterase (AChE) Staining, Nickel Enchanced DAB for Biocytin staining, and DAB for Biocytin staining.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 5 2020 Allen Institute for Brain Science 2020. DAB Solutions. protocols.io dx.doi.org/10.17504/protocols.io.bew7jfhn 2021-03-29 03:13:03
Electrode Internal Solution with Biocytin II
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bdn8i5hw 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 2 2020 Allen Institute for Brain Science 2020. Electrode Internal Solution with Biocytin II. protocols.io dx.doi.org/10.17504/protocols.io.bdn8i5hw 2021-03-29 03:13:01
Electrode Internal Solution with Biocytin II
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.bepnjdme 2021-03-29 03:13:09
PCR Prep from Glycerol Stock for IVT Reactions
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.brt2m6qe 2021-03-29 03:13:08
cDNA Synthesis
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.9geh3te 2021-03-29 03:13:11
Core Protocol for Serial Imaging of Fluorescently-Labeled Mouse Brain on the TissueCyte 1000 System
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.bdvmi646 2021-03-29 03:13:16
Mouse Whole Cell Tissue Processing for 10x Genomics Platform
 
Resource Report
Resource Website
Allen Institute for Brain Science 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 dx.doi.org/10.17504/protocols.io.bsfdnbi6 2021-03-29 03:13:18
Intraperitoneal Injection in an Adult Mouse
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bfzgjp3w BICCN, Allen Institute for Brain Science This protocol describes the general procedure used for intraperitoneal 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. Intraperitoneal Injection in an Adult Mouse. protocols.io dx.doi.org/10.17504/protocols.io.bfzgjp3w 2021-03-29 03:13:51
Multi-Patch Clamp Recording
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bcteiwje BICCN, Allen Institute for Brain Science This protocol describes the process to obtain multiple electrophysiological recordings from postnatal mouse and/or human brain slices. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Multi-Patch Clamp Recording. protocols.io dx.doi.org/10.17504/protocols.io.bcteiwje 2021-03-29 03:13:55
DAB Solutions
 
Resource Report
Resource Website
Allen Institute for Brain Science 10.17504/protocols.io.bdpdi5i6 BICCN, Allen Institute for Brain Science 3, 3´ Diaminobenzidine (DAB) is a commonly used chromogen for histological staining. In the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol and xylene. Peroxidase catalyzes hydrogen peroxide oxidation of substrate DAB. This SOP provides instructions to prepare four different formulations of DAB. DAB for Immunohistochemical staining (IHC), DAB for Acetylcholinesterase (AChE) Staining, Nickel Enchanced DAB for Biocytin staining, and DAB for Biocytin staining.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. DAB Solutions. protocols.io dx.doi.org/10.17504/protocols.io.bdpdi5i6 2021-03-29 03:14:02

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