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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
Case Processing SOP (Spleen)
 
Resource Report
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Marda Jorgensen 10.17504/protocols.io.bc3kiykw Human BioMolecular Atlas Program (HuBMAP) Method Development Community The purpose of this Standard Operating Procedure is to outline procedures for processing and storing spleen received for HuBMAP consortium assay and analysis. University of Florida 3 2020 Marda Jorgensen 2020. Case Processing SOP (Spleen). protocols.io dx.doi.org/10.17504/protocols.io.bc3kiykw 2021-03-29 03:08:40
High Resolution Imaging Mass Spectrometry Analysis using Bruker Daltonics Platforms
 
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Elizabeth Neumann, Jamie Allen, David Anderson, Danielle Gutierrez, Jeff Spraggins 10.17504/protocols.io.7gdhjs6 VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope:Acquire 10 μm spatial resolution imaging mass spectrometry (IMS) datasets of ~ 5mm2 regions of tissue.Expected Outcome:A series of ion images that can be used to visualize the distribution of lipids localizing to physiological regions within the human kidney. Vanderbilt University, Vanderbilt University, Vanderbilt University, Vanderbilt University, Vanderbilt University 1 2019 Elizabeth Neumann, Jamie Allen, David Anderson, Danielle Gutierrez, Jeff Spraggins 2019. High Resolution Imaging Mass Spectrometry Analysis using Bruker Daltonics Platforms. protocols.io dx.doi.org/10.17504/protocols.io.7gdhjs6 2021-03-29 03:07:52
PAS Staining of Fresh Frozen or Paraffin Embedded Human Kidney Tissue
 
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Jamie Allen, Jennifer Harvey, Maya Brewer, Mark De Caestecker, Jeff Spraggins 10.17504/protocols.io.4qngvve VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope:The PAS stain is used to demonstrate polysaccharides such as glycogen, and mucosubstances such as glycoproteins, glycolipids and mucins in tissues. It is used as a replacement for the H&E in kidney pathology.Expected Outcome:Intermyofibrillar Network…………………………..Pink to RoseGlycogen………………………………………………….…..Pink to RoseMyofibrils……………………………………………………..UnstainedType I fibers………………………………………………….LighterType II fibers…………………………………………………DarkerBlood Vessel walls & connective tissue…..Faintly Stained Vanderbilt University, Vanderbilt University, Division of Nephrology, Vanderbilt University Medical Center, Division of Nephrology, Vanderbilt University Medical Center, Vanderbilt University 1 2019 Jamie Allen, Jennifer Harvey, Maya Brewer, Mark De Caestecker, Jeff Spraggins 2019. PAS Staining of Fresh Frozen or Paraffin Embedded Human Kidney Tissue. protocols.io dx.doi.org/10.17504/protocols.io.4qngvve 2021-03-29 03:08:02
HuBMAP TMC-Florida/Zurich CODEX Modality Overview
 
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Marda Jorgensen, Jerelyn Nick, Franchesca Farris, Jesus Penaloza 10.17504/protocols.io.be9pjh5n Human BioMolecular Atlas Program (HuBMAP) Method Development Community This protocol is an overview of all of the protocols currently in use for the CODEX modality at HubMAP Tissue Mapping Center,TMC-Florida/Zurich. It includes protocols.io links to each of the individual protocols that make up this project workflow, from processing tissue and imaging the samples, to uploading the final data to the HuBMAP HIVE. University of Florida, University of Florida, University of Florida, University of Florida 2 2020 Marda Jorgensen, Jerelyn Nick, Franchesca Farris, Jesus Penaloza 2020. HuBMAP TMC-Florida/Zurich CODEX Modality Overview. protocols.io dx.doi.org/10.17504/protocols.io.be9pjh5n 2021-03-29 03:08:05
Post-IMS Autofluorescence Microscopy
 
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Elizabeth Neumann, Jamie Allen, Heath Patterson, Danielle Gutierrez, Jeff Spraggins 10.17504/protocols.io.879hzr6 VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope: Obtain autofluorescence microscopy images of tissues after IMS analysisExpected Outcome: A brightfield and green autofluorescence microscopy image of the tissue section that enables registration and correlation of different imaging modalities on a pixel by pixel basis. Vanderbilt University, Vanderbilt University, Vanderbilt University, Vanderbilt University, Vanderbilt University 1 2020 Elizabeth Neumann, Jamie Allen, Heath Patterson, Danielle Gutierrez, Jeff Spraggins 2020. Post-IMS Autofluorescence Microscopy. protocols.io dx.doi.org/10.17504/protocols.io.879hzr6 2021-03-29 03:11:20
Lightsheet Tissue Intake - Photodocumentation and Tracking
 
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Seth Currlin, Marda Jorgensen 10.17504/protocols.io.bef6jbre Optical Clearing of Tissue, Human BioMolecular Atlas Program (HuBMAP) Method Development Community In order to track tissue position and orientation each sample is photographed when received for lightsheet microscopy (Figure 1, A). Large samples (>1 cm3), such as the spleen in Figure 1, are cut into 2 mm sections (Figure 1, B) to facilitate tissue clearing, staining, and three-dimensional imaging. These tissues are batch processed and withdrawn from the lipid clearing pipeline as needed (Figure 1, C); at this time a novel identifier is assigned to the sample. University of Florida, University of Florida 1 2020 Seth Currlin, Marda Jorgensen 2020. Lightsheet Tissue Intake - Photodocumentation and Tracking. protocols.io dx.doi.org/10.17504/protocols.io.bef6jbre 2021-03-29 03:11:24
UF H&E Staining
 
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Marda Jorgensen, Franchesca Farris 10.17504/protocols.io.beamjac6 Human BioMolecular Atlas Program (HuBMAP) Method Development Community Hematoxylin and Eosin stains are the prefered method for histopathologic assessment of tissue sections.Hematoxylin is used to illustrate nuclear detail in cells. The resulting intensity and shade of the dye in the cells is primarily reliant on the length of time the sample spends in hematoxylin. The cytoplasmic component of the section is identified by Eosin, which is the most commonly used counterstain in histology. It's varying shades of pink and red provide an exceptional contrast between the nuclear and cytoplasmic areas of interest. University of Florida, University of Florida 1 2020 Marda Jorgensen, Franchesca Farris 2020. UF H&E Staining. protocols.io dx.doi.org/10.17504/protocols.io.beamjac6 2021-03-29 03:11:51
Protocol for Freezing in OCT
 
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Shin Lin, Yiing Lin 10.17504/protocols.io.bpx7mprn Human BioMolecular Atlas Program (HuBMAP) Method Development Community University of Washington, Washington University, Saint Louis 1 2020 Shin Lin, Yiing Lin 2020. Protocol for Freezing in OCT. protocols.io dx.doi.org/10.17504/protocols.io.bpx7mprn 2021-03-29 03:08:49
DAB Staining of FFPE Slides
 
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Jerelyn Nick, Franchesca Farris, Marda Jorgensen 10.17504/protocols.io.bdiei4be Human BioMolecular Atlas Program (HuBMAP) Method Development Community Standard IHC staining, such as this DAB protocol, is included in the validation of new antibodies at the University of Florida Tissue Mapping Center, TMC-Florida/Zurich. This procedure includes all of the reagents necessary to perform a DAB based immunohistochemistry reaction. DAB Substrate (3,3’-diaminobenzidine) produces a brown reaction product in the presence of peroxidase (HRP) enzyme. University of Florida, University of Florida, University of Florida 1 2020 Jerelyn Nick, Franchesca Farris, Marda Jorgensen 2020. DAB Staining of FFPE Slides. protocols.io dx.doi.org/10.17504/protocols.io.bdiei4be 2021-03-29 03:08:51
Autofluorescence Microscopy Data Acquisition
 
Resource Report
Resource Website
Elizabeth Neumann, Heath Patterson, Jamie Allen, Maya Brewer, Mark De Caestecker, Danielle Gutierrez, Jeff Spraggins 10.17504/protocols.io.7e3hjgn VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope:Obtain autofluorescence microscopy images of tissues.Expected Outcome:An RGB autofluorescence microscopy image of the tissue section that enables registration and correlation of different imaging modalities on a pixel by pixel basis. Vanderbilt University, Vanderbilt University, Vanderbilt University, Vanderbilt University, Division of Nephrology, Vanderbilt University Medical Center, Vanderbilt University, Vanderbilt University 1 2019 Elizabeth Neumann, Heath Patterson, Jamie Allen, Maya Brewer, Mark De Caestecker, Danielle Gutierrez, Jeff Spraggins 2019. Autofluorescence Microscopy Data Acquisition. protocols.io dx.doi.org/10.17504/protocols.io.7e3hjgn 2021-03-29 03:08:54
HuBMAP Tissue Sectioning for FFPE Specimens
 
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Leigh Propper, Marda Jorgensen 10.17504/protocols.io.bahdib26 Human BioMolecular Atlas Program (HuBMAP) Method Development Community This method details our microtomy (sectioning) process for reasearch specimens involved with HuBMAP.This process follows the microtomy process that follows after receiving, processing, and embedding of research specimens into formalin fixed, paraffin embedded tissue blocks. University of Florida, University of Florida 2 2019 Leigh Propper, Marda Jorgensen 2019. HuBMAP Tissue Sectioning for FFPE Specimens. protocols.io dx.doi.org/10.17504/protocols.io.bahdib26 2021-03-29 03:09:26
HuBMAP - Tissue Processing with Paraffin Infiltration
 
Resource Report
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Marda Jorgensen, Franchesca Farris, Jerelyn Nick 10.17504/protocols.io.8nzhvf6 Human BioMolecular Atlas Program (HuBMAP) Method Development Community The purpose of this Standard Operating Procedure (SOP) is to outline procedures for the automated processing of formalin fixed HuBMAP specimens for paraffin embedding. University of Florida, University of Florida, University of Florida 1 2020 Marda Jorgensen, Franchesca Farris, Jerelyn Nick 2020. HuBMAP - Tissue Processing with Paraffin Infiltration. protocols.io dx.doi.org/10.17504/protocols.io.8nzhvf6 2021-03-29 03:07:31
Case Processing SOP (Spleen)
 
Resource Report
Resource Website
Marda Jorgensen 10.17504/protocols.io.bbcuiiww Human BioMolecular Atlas Program (HuBMAP) Method Development Community The purpose of this Standard Operating Procedure is to outline procedures for processing and storing spleen received for HuBMAP consortium assay and analysis. University of Florida 2 2020 Marda Jorgensen 2020. Case Processing SOP (Spleen). protocols.io dx.doi.org/10.17504/protocols.io.bbcuiiww 2021-03-29 03:07:43
FFPE Tissue Sectioning for Staining
 
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Marda Jorgensen 10.17504/protocols.io.8kvhuw6 Human BioMolecular Atlas Program (HuBMAP) Method Development Community Method for sectioning paraffin blocks for staining University of Florida 1 2019 Marda Jorgensen 2019. FFPE Tissue Sectioning for Staining. protocols.io dx.doi.org/10.17504/protocols.io.8kvhuw6 2021-03-29 03:10:43
Cell DIVE™ Platform | Ab Conjugation: Initial Conjugation & Scale up Conjugation
 
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Anup Sood, Eric Williams, Liz McDonough 10.17504/protocols.io.bp55mq86 Human BioMolecular Atlas Program (HuBMAP) Method Development Community, GE Research ScopeThe purpose of this protocol is to conjugate purified antibodies to Cy dyes. Antibody conjugation involves combining dyes with antibodies to achieve a fluorescently labeled antibody. Each antibody will be labeled with a molar equivalent of dye; the quantitative output of this is deemed D/P. In the initial conjugation step, each antibody will be labeled with differing D/P levels and validated to determine which level is optimal. GE Research, GE Research, GE Research 1 2021 Anup Sood, Eric Williams, Liz McDonough 2021. Cell DIVE™ Platform | Ab Conjugation: Initial Conjugation & Scale up Conjugation. protocols.io dx.doi.org/10.17504/protocols.io.bp55mq86 2021-03-29 03:10:45
Automatic Deposition of DAN Matrix using a TM Sprayer for MALDI Analysis of Lipids
 
Resource Report
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Jamie Allen, Elizabeth Neumann, Jeff Spraggins 10.17504/protocols.io.4srgwd6 VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope:To describe the procedure for spraying tissue sections with DAN for imaging lipids.Expected Outcome:Slides should be coated with DAN. Tissue sections should be imaged within 24 hours of matrix application. Vanderbilt University, Vanderbilt University, Vanderbilt University 1 2019 Jamie Allen, Elizabeth Neumann, Jeff Spraggins 2019. Automatic Deposition of DAN Matrix using a TM Sprayer for MALDI Analysis of Lipids. protocols.io dx.doi.org/10.17504/protocols.io.4srgwd6 2021-03-29 03:10:50
Imaging Mass Cytometry Antibody Staining
 
Resource Report
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Michelle Daniel, Marda Jorgensen 10.17504/protocols.io.bfz8jp9w Human BioMolecular Atlas Program (HuBMAP) Method Development Community The following SOP describes procedures required for the antibody staining of formalin fixed,paraffin-embedded (FFPE) samples. In this SOP, the term “samples” refers to FFPEtissue samples. University of Zürich, University of Florida 1 2020 Michelle Daniel, Marda Jorgensen 2020. Imaging Mass Cytometry Antibody Staining . protocols.io dx.doi.org/10.17504/protocols.io.bfz8jp9w 2021-03-29 03:10:56
HuBMAP Tissue Sectioning for FFPE Specimens
 
Resource Report
Resource Website
Leigh Propper, Marda Jorgensen 10.17504/protocols.io.93qh8mw Human BioMolecular Atlas Program (HuBMAP) Method Development Community This method details our microtomy (sectioning) process for reasearch specimens involved with HuBMAP.This process follows the microtomy process that follows after receiving, processing, and embedding of research specimens into formalin fixed, paraffin embedded tissue blocks. University of Florida, University of Florida 1 2019 Leigh Propper, Marda Jorgensen 2019. HuBMAP Tissue Sectioning for FFPE Specimens. protocols.io dx.doi.org/10.17504/protocols.io.93qh8mw 2021-03-29 03:11:02
Automated, Rapid Preparation of Tissue Sections for Proteomic Analysis
 
Resource Report
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Jamie Allen, Jeff Spraggins, Danielle Gutierrez 10.17504/protocols.io.67nhhme VU Biomolecular Multimodal Imaging Center, Human BioMolecular Atlas Program (HuBMAP) Method Development Community Scope:To describe the procedure for the lysis, reduction/alkylation, trypsin digestion, and clean-up of tissue. Lysis will cover the lysing of tissue and protein concentration. Acetone precipitation will cover the precipitation of proteins. Digestion will cover the process for digesting 100 µg of protein using the Agilent AssayMap Bravo Robot and Promega Rapid Trypsin/LysC. The clean-up of the cells will cover the desalting process on the Agilent AssayMap Bravo and the subsequent preparation of the samples for LC/MS peptide analysis.Expected Outcome/Data:Cell samples lysed, digested, and desalted for analysis on MS instrument. Samples to be analyzed within one or two days of desalting. Vanderbilt University, Vanderbilt University, Vanderbilt University https://www.thermofisher.com/order/catalog/product/23225 2 2019 Jamie Allen, Jeff Spraggins, Danielle Gutierrez 2019. Automated, Rapid Preparation of Tissue Sections for Proteomic Analysis. protocols.io dx.doi.org/10.17504/protocols.io.67nhhme 2021-03-29 03:11:09
Donor Acceptance Criteria for TMC Florida/Zurich HuBMAP Inclusion
 
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Marda Jorgensen 10.17504/protocols.io.bipykdpw Human BioMolecular Atlas Program (HuBMAP) Method Development Community This document outlines the required criteria for inclusion for donors of thymus, lymph nodes and spleen in the HuBMAP project. University of Florida 1 2020 Marda Jorgensen 2020. Donor Acceptance Criteria for TMC Florida/Zurich HuBMAP Inclusion. protocols.io dx.doi.org/10.17504/protocols.io.bipykdpw 2021-03-29 03:12:21

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