Immunomics workflow at TII is a cogent platform where various high end technologies come together in a comprehensive manner to deliver maximum insight into patient samples. Technologies are deployed in a stepwise manner such that sample interrogation occurs in a methodical fashion. Mass cytometry is often employed as a hypothesis building tool due to its broad capability and versatility of utilising several antibody combinations as a panel. The CyTOF procedure is used for deep immunophenotyping the patient samples to characterise sub-populations of interest. Once populations of interest are identified, specific cell sub sets are further studied by sorting them by flowcytometry. Isolated sub sets can then be studied using a combinatorial approach using several molecular and functional analyses. Briefly, the workflow for immunomics involves:
•Patient samples are interrogated using mass cytometry (CyTOF) that allows for up to 41 individual markers to be assessed on single cells. This allows for deep immunophenotyping the patient samples to characterise sub-populations of interest. •Specific immune sub-populations for further study are isolated through various cell sorting approaches •Immune cell clusters of interest are dissected for function and underlying molecular and pathway analyses either at the population or single cell level•Multiplexed tissue phenotyping using Vectra methodology allows for studying tissue microenvironment contribution and modulation of immune responses in disease while preserving cellular and tissue geography
•Cell free/ secreted molecules participating in immune modulation can be studied using multiplexed approaches (Luminex) or ELISA
•Transcriptomic analyses either on cell clusters or single cells and Nanostring technology allow for high throughput molecular information of the immune landscape
A robust bioinformatics expertise and infrastructure exist in-house to support development of analyses pipelines and statistical analyses.We also have an in house high volume bio-banking capacity currently capable of storing up to 45000 vials under LN2 vapour. We, along with other participating entities in the SingHealth cluster, have recently been ISO 9001:2015 certified.