HuBMAP | GE HealthCare/UPitt Cell DIVE™ and scRNA Modality Overview
Liz McDonough
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.
Steps
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
Perform skin digestion and scRNA analysis.
Cell DIVE Multiplexed Immunofluorescence (MxIF)
Prepare formalin fixed and paraffin embedded (FFPE) blocks and tissue sections.
HuBMAP | Formalin Fixation and Paraffin Embedding of Tissue Samples
Deparaffinize and rehydrate slides.
Characterize antibodies (primary/secondary, direct conjugates, and zenon labelled antibodies) and determine any antigen effects from the Cell DIVE dye inactivation process.
Cell DIVE™ Platform | Antibody Characterization for Multiplexing
Prepare direct conjugates for study.
Cell DIVE™ Platform | Antibody Purification Chemistry
Cell DIVE™ Platform | Ab Conjugation: Initial Conjugation & Scale up Conjugation
Perform Cell DIVE™ multiplexed data acquisition on the final cohort.
