Mar 21, 2024
HuBMAP | GE HealthCare/UPitt Cell DIVE™ and scRNA Modality Overview
- 1GE HealthCare
Protocol Citation: Liz McDonough 2024. HuBMAP | GE HealthCare/UPitt Cell DIVE™ and scRNA Modality Overview. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v92ekzl3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 21, 2024
Last Modified: March 22, 2024
Protocol Integer ID: 97070
Funders Acknowledgement:
HuBMAP
Grant ID: U54 AR081775-01
Disclaimer
The protocols.io team notes that research involving animals and humans must be conducted according to internationally-accepted standards and should always have prior approval from an Institutional Ethics Committee or Board.
Abstract
This is an overview of all protocols currently in use for the 2024 GE HealthCare/UPitt collaboration for the Human BioMolecular Atlas Program (HuBMAP). It includes links to each of the individual protocols that make up this project workflow.
Obtaining Donors and Tissue Blocks
Obtaining Donors and Tissue Blocks
Confirm donor acceptance criteria for inclusion.
5 mm skin biopsies obtained from donor subjects are divided into two parts, with one half for formalin fixed and paraffin embedded (FFPE) processing and the other half for skin digestion for further scRNA analysis.
scRNA Analysis
scRNA Analysis
Cell DIVE Multiplexed Immunofluorescence (MxIF)
Cell DIVE Multiplexed Immunofluorescence (MxIF)
Prepare formalin fixed and paraffin embedded (FFPE) blocks and tissue sections.
Characterize antibodies (primary/secondary, direct conjugates, and zenon labelled antibodies) and determine any antigen effects from the Cell DIVE dye inactivation process.
Prepare direct conjugates for study.
Perform Cell DIVE™ multiplexed data acquisition on the final cohort.
Note
Staining is done using the Leica Bond MAX and images are acquired on the IN Cell Analyzer 2200.