Mass cytometry (MC) is a powerful research tool enabling high dimensional analysis of single cells in suspension and, following laser ablation, from tissue sections. By replacing fluorophore-conjugated antibodies with heavy metal-conjugated antibodies, MC enables the simultaneous detection of over 40 parameters with minimal overlap between reporters. However, adopting this technology requires significant investment, technical expertise and knowledge of immunology, which is prohibitive for many users. Operating independently, a researcher would have to purchase a large commercial panel to perform even preliminary MC experiments.
At the Ramaciotti Facility for Human Systems Biology (RFHSB), a joint initiative between The Sydney Cytometry Facility, the University of Sydney and the Centenary Institute, we have developed a streamlined, affordable system to facilitate the use of MC. The RFHSB has produced over 600 ready-to-use metal-conjugated antibodies for use in solution mode MC and titrated these in relevant tissues. Users can purchase these quality controlled ready-to-use antibodies in a per test volume, reducing the cost and time investment substantially.
While antibody clones used successfully for solution mode MC and fixed tissue sections overlap, <90% of clones translate for use in formalin fixed paraffin embedded tissue (FFPE). To address this, the RFHSB collaborated with researchers from various fields to design panels, conjugate and validate new reagents for imaging MC (IMC). Knowledge from collaborators regarding FFPE antibody clones that are effective in their tissues is used to inform the choice of clones for in-house conjugation. These reagents are tested on target tissues in collaboration with users. We have validated >50 reagents for use in FFPE tissue and >60 for use in frozen tissue sections from various sources, facilitating the use of IMC for a growing number of research groups in Australia. This has enabled us to reduce costs associated with IMC and develop reagents not commercially available.