HuBMAP | GE/ University of Washington Cell DIVE™ Modality Overview

Liz McDonough

Published: 2022-04-08 DOI: 10.17504/protocols.io.3byl4bwmzvo5/v1
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Abstract

This is an overview of all protocols currently in use for the GE/University of Washington 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

1.

Protocol for Tissue Collection from Organ Procurement Organization

2.

Prepare paraffin blocks and FFPE sections from tissue samples.

HuBMAP Tissue Preservation Protocol

3.

Deparaffinize and rehydrate slides.

Cell DIVE™ Platform | Slide Clearing and Antigen Retrieval

4.

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

5.

Prepare direct conjugates for study.

Cell DIVE™ Platform | Antibody Purification Chemistry

Cell DIVE™ Platform | Ab Conjugation: Initial Conjugation & Scale up Conjugation

6.

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 Leica Cell DIVE imager.

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