OVERVIEW OF SERVICES 

Immune Monitoring and Flow Cytometry Shared Resource (IMFC SR) provides state-of-the-art immune monitoring services for clinical and translational studies to characterize the immune system. IMFC SR is a 24 X 7 Flow cytometry instrumentation facility providing multiparametric spectral (up to 34 colors) and conventional flow cytometry (14 colors) assay design and hi-dimensional data analysis tools (tSNE, UMAP, TriMAP, Flow-SOM, etc.). We offer staff-assisted cell sorting (bulk and single-cell), human and mouse primary cells, cell lines, exosomes, and nuclei sorting. Sorters are capable of up to 4 and 6-way sort. For cytokine analysis, we offer the Luminex-200 platform (which can detect up to 65 protein analytes). Bulk gene expression analysis using the NanoString platform, single-cell RNA sequencing using libraries created with 10X genomics technology, xCELLigence Real-Time Cell Analysis (RTCA) assay, and sample processing (tumor tissue, whole blood, serum, and plasma). We provide consultation for assay design and clinical trials.

Acknowledgment of Immune Monitoring and Flow Cytometry SR

The Immune Monitoring and Flow Cytometry Shared Resource of Rutgers Cancer Institute of New Jersey is a specialized service facility that supports the cancer research efforts of our members. Please remember to acknowledge the valuable services the Shared Resource provides in your research papers, publications, and grant applications. Use the following acknowledgment statement:

"Services, results, and products in support of the research project were generated by the Rutgers Cancer Institute of New Jersey Comprehensive Genomics Shared Resource, supported, in part, with funding from the NCI-CCSG P30CA072770-5920."

Please consider including the names of individuals from the Shared Resource if they provided any intellectual input or additional effort.

NIH Public Access Policy: Publications from services provided by this Shared Resource must comply with the NIH Public Access Policy by submitting your paper to PubMed Central.

Go to: https://publicaccess.nih.gov/submit_process.htm for Pubmed Central's submission instructions.

EQUIPMENT

Cell Sorter

1. Cytek Aurora Spectral Cell Sorter

2. BD Influx Cell Sorter

Flow Cytometry Analyzers

1. Cytek Aurora Spectral Cell analyzer with 96 well plate loader. (Blue, Violet, UV, and Red Laser- detects up to 34 Flurochromes).

2. Attune 4L NxT Cytometer with additional 384, 96 well plate loader (Blue, Violet, Yellow Green, and Red lasers - detects up to 14 Flurochromes).

 3. Attune 3L NxT Cytometer (Blue, Violet, Red lasers - detects up to 11 Flurochromes).

Luminex 200 System

Serum, plasma, culture supernatant cytokine/chemokine/soluble cellular factor/growth factor analysis is performed on the Luminex 200 Platform. This instrument can perform multiplex analysis of up to 100 analytes in as little as 50 microliters of serum or cell culture supernatant. The typical dynamic range for each analyte is greater than or equal to 3.5 logs for extremely sensitive and reproducible interrogation.

NanoString nCounter Max System

Targeted gene expression analysis is performed using a NanoString nCounter Max System. Combining an automated prep station and digital analyzer unit, this system can perform multiplex analysis on up to 800 RNA, DNA, or protein targets in up to 12 samples per run. The nCounter instruments provide a simple and cost-effective approach that can be used with various sample types, including FFPE-processed tissue.

10X Genomics Chromium Controller

The Immune Monitoring and Flow Cytometry Shared Resource provides assays on the 10X Chromium Controller for single-cell gene expression. The Chromium controller uses advanced microfluidics to process up to 8 samples into single-cell gel emulsions ready for RNA barcoding and cDNA amplification. The trained technical staff in the Shared Resource create both general and targeted gene expression libraries from this amplified, barcoded cDNA for downstream sequencing and analysis. 

2100 Bioanalyzer System

The 2100 Bioanalyzer System provides automated electrophoresis for quality control of biomolecules. This chip-based assay can perform RNA, DNA, and protein analysis. Compared to traditional electrophoresis, this system allows for lower sample input (1 microliter for Nucleic Acids, 4 microliters for protein) and significantly reduces the analysis times. Output from the instrument includes assessment of sizing, quantitation, integrity, and purity of RNA, DNA, and proteins.

Invitrogen Qubit 4 Fluorometer 

The Invitrogen Qubit 4 Fluorometer utilizes assays based upon dyes that bind selectively to DNA, RNA, or protein, making it more specific than UV-absorbance-based assays. It quickly measures integrity and quality and quantifies each sample in under 3 seconds. With increased sensitivity, it can reliably detect as little as 10 picrograms/microliter of DNA. 

xCELLigence RTCA 

The Agilent xCELLigence Real-Time Cell Analysis (RTCA) instrument uses label-free cellular impedance to continuously monitor cell health, behavior, and function with high accuracy, sensitivity, and reproducibility. It can simultaneously run six 96-well electronic microplates (E-Plate 96), which is ideal for cell health characterization, immune cell-mediated killing, viral cytopathic effects, cytotoxicity, cell adhesion, cell barrier function (TEER), cell signaling (e.g., GPCR) and other related applications. The instrument operates in a standard CO2 cell culture incubator, and the control unit is housed outside the incubator. Simply plate cells and begin monitoring cell behavior to obtain real-time kinetic data for assay windows that stretch from seconds to days. Online data acquisition and offline data analysis are easy to perform using RTCA Software Pro, supporting FDA 21 CFR Part 11 compliance.

Sample Processing and Storage

• PBMC isolation from whole blood

• Serum isolation from whole blood

• Tumor-infiltrating lymphocyte (TIL) isolation from tissue

• RNA/DNA isolation from cells or tissue

• ccfDNA isolation from whole blood

• RNA isolation from FFPE tissue blocks

• Custom isolation/dissociation available upon consultation

CONTACT INFORMATION

LINKS AND RESOURCES

Immune Monitoring and Flow Cytometry Shared Resource Website

Rutgers iLab User Manuals

Rutgers iLab Video Tutorials

Rutgers iLab FAQs

Contacts