HuBMAP | GE/URMC/SCH Cell DIVE™ Modality Overview
Gloria S Pryhuber, Heidie Huyck, Cory Poole, Lisa Lowery, gail.deutsch, Christine Surrette
Abstract
This is an overview of all protocols currently in use for the GE/URMC/SCH Cell DIVE collaboration for the Human BioMolecular Atlas Program (HuBMAP). It includes links to each of the individual protocols that make up this project workflow.
Steps
Confirm donor acceptance criteria for inclusion.
URMC | Donor Acceptance Criteria for GE/URMC HuBMAP Inclusion
URMC | COVID+ Donor Acceptance Criteria for GE/URMC HuBMAP Inclusion
Prepare paraffin blocks and FFPE sections from tissue samples.
URMC | Whole Lung and Lobe Processing - Formalin Fixation and Gross Sectioning of Tissue Samples
URMC | Parrafin Embedding and Sectioning of FFPE Lung Tissue
Deparaffinize and rehydrate slides.
Cell DIVE™ Platform | Slide Clearing and Antigen Retrieval
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
Cell DIVE™ Platform | Antibody Staining & Imaging
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.
