Platform Assays
Our Services
Using the latest technology, the I3H Platform Assays division offers comprehensive biospecimen services to preapproved researchers:
- Customized project management
- Biospecimen processing from low minimum sample volumes
- Molecular and cellular immunological assays
- Data analysis
- Serology
Continually upgrading and expanding, I3H is currently developing automated workflow systems to increase efficiency, fine-tune quality, optimize blood product utilization, and further decrease required sample volume. We invite you to tell us about your needs and inquire about any of our services.
Project Management and Consultation
I3H offers individualized guidance on study design and set-up, including all workflow considerations — from standard operating procedure (SOP) selection and IRB review, through sample collection, processing customization, assay selection, additional quality control (QC) metrics, and data analysis.
Biospecimen Processing
Our dedicated full-time staff offers same-day in-house processing of whole blood samples for extraction of viable human peripheral blood mononuclear cells (PBMCs) and nonviable products (i.e., serum, plasma, DNA, buffy coats). We also offer same-day staining for mass cytometry assays from the same draw. Only 300 µL of fresh whole blood is required for the MDIPA mass cytometry service. I3H guarantees >90% cell viability for samples processed within 24 hours and follows rigorous preservation protocols to optimize cell recovery for downstream processes. We adhere to strict standard operating procedures to assure regulatory compliance.
Viable cell processing services include the following:
- Continuous tracking and quality control of PBMC recovery/yields
- Complete blood count (CBC) with differential, available for all EDTA-collected samples using a Beckman Coulter DxH520 hematology analyzer, which requires only 17 µL of whole blood
- Flow cytometry-based human leukocyte antigen (HLA) typing, which facilitates MHC Class I tetramer assays
- Measurement of CMV IgG and EBV-EBNA1 IgG in serum/plasma
- Cryopreservation and storage of live PMBCs, serum, and plasma
Biobanking/Cryopreservation
The Platform Assays processing unit features centrally monitored (Aegis Scientific remote monitoring system) -80° C and liquid nitrogen freezers for readily accessible short- and long-term sample storage as well as internally monitored refrigerators for storage of samples, antibodies, and reagents.
Advanced, Accurate Sample Tracking
I3H works closely with the University of Pennsylvania’s Laboratory Information Management LabVantage team. Our LabVantage specialists support more than 60 separate research teams, 700 users, and 325 studies. They have tracked more than 1.5 million samples.
All samples in our processing unit are barcoded to facilitate tracking from blood-draw scheduling through transport to storage and retrieval. Our software enables us to link assay information to individual samples, along with the specific reagents used in each.
In addition to precise sample tracking, continuous collection of standardized, detailed processing attributes and other experiment specifics provides vital information for data interpretation and future cross-referencing needed for potential metadata linkage.
For questions, consultations, and study initiation requests, see more information.
Immunological Assays and Analysis
We provide high-quality, high-dimensional immune profiling and analysis that enables comparison of individuals across studies. Human biology demands that assaying be performed in accordance with rigid quality control standards to minimize technical noise. I3H submits every assay batch to rigorous QC processes, capturing and documenting observable technical variations to maximize results for clear interpretation.
- We provide QC reports with every set of data files acquired.
- We run internal control samples with each data acquisition to monitor instrument performance over time.
- We document which lots of reagent are used for each step of the assay and test and analyze new lots to ensure consistency.
- We document all unavoidable reagent or procedural variations and use that information for sample metadata when interpreting results.
Additionally, we are in the process of developing novel batch correction methods through our Informatics division.
Mass Cytometry (CyTOF) Services
Mass cytometry, also known as cytometry by time-of-flight (CyTOF®), is a state-of-the-art technology that tags antibodies with heavy-metal isotopes to measure protein expression on hundreds of thousands of individual cells.
CyTOF MDIPA Assays
For high-dimensional profiling of whole blood and PMBCs, I3H performs the Maxpar®â€¯Direct™ Immune Profiling Assay™ (MDIPA), using the CyTOF XT™ — Standard BioTools’ next-generation mass cytometer. With this powerful system, I3H can offer investigators the kind of crucial, reproducible data they need.
- Measurement of 30 preset protein markers on more than 40 distinct immune cell populations
- Optional customization of an additional 15 markers
- Staining of fresh whole blood to capture granulocytes, in addition to mononuclear cells, with a required sample of only 300 µL and the flexibility of either EDTA or heparin collection
- Staining of fresh or cryopreserved PBMCs, from a single cell to 3 million cells
- Preset add-on panels to further interrogate the following:
- Acute myeloid leukemia (AML)
- Myeloid/neutrophilic populations
- Customized metal-isotope antibody conjugation
Flow Cytometry Services
Flow cytometry uses fluorescent dye on antibodies to measure protein expression on individual cells.
I3H currently offers the following:
- Predesigned and validated immunophenotyping panels:
- General Immunophenotyping (17 markers)
- Monocyte/DC focused (35 markers)
- AML (25 markers)
- Customized panel design and validation (upon consultation/validation)
We are in the process of building, validating, and standardizing population-specific flow cytometry panels.
Cytometric Data Analysis
Tier 1 CyTOF
Tier 1 CyTOF assay analysis provides investigators with an autogenerated summary of immune cell populations (>40). Study samples can be compared to more than 500 other individuals from over 1,200 samples we’ve acquired to date. The dataset comparison spans health conditions affecting the immune system, such as autoimmunity, cancer, infectious diseases, and vaccinations, and includes a healthy population reference.
Tier 2 CyTOF and Flow Cytometry
For both Tier 2 CyTOF and flow cytometry assays, I3H performs unsupervised method analysis of high-dimensional data and statistical testing. We also offer reference analysis to previous openly available or I3H-generated databases/studies.
Serology Services
I3H performs a monochromator-based enzyme-linked immunosorbent assay (ELISA) using the Agilent BioTek Epoch microplate spectrophotometer to evaluate serum/plasma measures of IFN-γ and IL-6.
For More Information
Study Initiation and Consultation
We are eager to assist you with your research. Pre-approval is required, and I3H will only approve new study requests for which we can confirm resource availability in advance.
Your request must meet the following criteria, in accordance with our vision and procedures.
- Any new study must have a clear goal of clinical impact, such as integration into the electronic medical record, development of new diagnostic tests, initiation of a new clinical trial, etc.
- Each project must have a champion that demonstrates commitment to clinical development beyond academic publications.
- Studies must be well powered with achievable parameters and time frame for completion. Reasonable effort is required on all sides.
- Investigators must have sufficient funding, demonstrated in advance. Studies requiring additional protocols or late processing will be charged accordingly.
To initiate a study or for consultation, please contact Damian Maseda, PhD.
Platforms Assays and Analysis
Please direct questions about specific assays and analysis to one of the following people.
Position | Name/Email |
---|---|
Scientific Director of Immune Health Laboratories | Damian Maseda, PhD |
Assistant Director of Platform Assays | Michelle McKeague, PhD |
Mass Cytometry Lead | Shwetank Verma, PhD |
Flow Cytometry Lead | Ayman Rezk, PhD |
Senior Data Analyst | Matei Ionita, PhD |