Our team specializes in immunological assay development with a strong focus on collaboration, data quality, and reproducibility in the following areas:
- Multiparametric flow cytometry analysis (Cytek Aurora)
- Antibody level analysis (e.g., rabbit ATG)
- Fluorescence-activated cell sorting (BD FACSMelody)
- Large-scale analysis for secreted biomarkers (Olink Platform)
- Single-cell cytokine secretion analysis (IsoPlexis)
- PK/PD modeling (in collaboration with the LCMS PK facility; CLIA approved, including fludarabine, busulfan, clofarabine, melphalan
- Coming soon: e.g., TCRseq, transcriptomics (in collaboration with other labs)
Multiparametric flow cytometric analysis
Multiparametric flow cytometric analysis using the Cytek Aurora cell analyzer provides comprehensive phenotyping of immune populations and cellular signatures. Expanded leukocyte typing is performed for the purpose of in-depth monitoring of checkpoint-related markers, regulatory T cells, NK cells, myeloid cells, and other cell types.
The Cytek Aurora cell analyzer has five lasers (355, 405, 488, 561, 640 nm), three scattering channels, and 64 fluorescence channels, as well as spectral detection technology to facilitate the simultaneous detection of more than forty discrete multiplexed probes. This analyzer measures the entire emission spectra of the fluorescent dyes excited by multiple lasers in order to allow the separation of fluorochromes with very close emission profiles. The Aurora’s laser beam profile and height are optimized for the detection of submicron particles and allows for optimal performance at high sample flow rates.
Examples of our advanced immune monitoring panels:
The IDMS provides cell sorting services using the FACSMelody cell analyzer for the separation and single-cell sorting of specific immune populations for the purpose of downstream applications and analysis.
Our FACSMelody cell sorter is equipped with three lasers (405, 488, and 561 nm) that allow for the detection of 11 parameters. It is capable of 4-way sorting with a 100 µM nozzle. It is enclosed in a BSL2 biosafety cabinet to ensure safe sorting of live human cells and cells exposed to infectious agents. The instrument’s automated software setup is designed to speed up and simplify operation.
The IDMS uses the Olink technology platform for discovery proteomics of cell therapy samples to detect soluble biomarkers.
The Olink proteomics platform is a disease-related large-scale biomarker analysis platform, utilizing a matched pair of antibodies linked to unique oligonucleotides for each biomarker of interest. Olink employs proximity extension assay (PEA) technology, allowing hybridization of matching nucleotides in close proximity to each other, followed by the annealing and amplification steps of PCR. This highly multiplexed and high throughput technology allows the simultaneous measurement of 92 disease-related biomarkers within 1 ul of sample.
Single cell analysis
The IsoPlexis platform, a proteomic 32-plex antibody-barcoded chip analysis technology, provides functional phenotypic analysis of single cells (CAR-T cells, etc.). The instrument, in combination with the IsoCode chips, can measure 30+ cytokines on single cell bases. Each chip can deliver cytokine profiles of about 1000 individual cells.
The IDMS evaluates viable cell subsets by polyfunctional profiling (the ability of cells to secrete ≥2 cytokines) and determines their polyfunctional strength index (PSI), measuring the percentage of polyfunctional cells in the sample, multiplied by the intensities of the secreted cytokines.