MASS CYTOMETRY

Generate new immune-related hypotheses using high parameter exploration

Using Fluidigm’s Helios®, HIMSR is equipped to help you generate new immune-related hypotheses, identify or phenotype immune cells in your study, and understand their signaling pathways via the detection of cytokines. The Helios CyTOF is a mass cytometer capable of high-parameter single-cell analysis. Rather than using fluorescence like flow cytometry, it uses mass spectrometry to detect heavy metal-conjugated antibodies. With this instrument, we can analyze up to 40 different external and internal cellular markers for you. We have developed antibody panels or can help you develop new antibody panels to analyze your cells. Log into iLab for service fees and panels; contact Angie Minic for a quote.

The CyTOF: This technology was developed for samples in single cell suspension (peripheral blood, PBMCs, disaggregated tissue). The cells can be live, fixed, or frozen - there are not many limitations assuming the antibodies specifically label your sample. Using special sample preparation, signaling and cytokine production can be interrogated. The sample is stimulated on the day of collection for either 30 min (for signaling assays) or 4-6 hours (to assay cytokine production). For signaling assays, we use fresh samples because frozen/thawed samples do not perform as well. 

Barcoding: To reduce variability in staining and the instrument, we barcode up to 20 samples per experiment so that they are stained and run together in one combined sample. Barcoding requires permeabilizing and labeling each individual sample with a heavy metal “code” prior to pooling the samples so that data for each sample can be separately analyzed after acquisition.

Metal antibody panels: We have developed a phenotyping panel that identifies a broad variety of immune cell types. The antibodies in this panel work on both fixed and live cells, and have been tested on whole blood and PBMCs. If a standard antibody is not available for your marker(s) of choice, we will work with you to develop a custom human antibody panel in your lab or in the HIMSR. We will customize signaling and cytokine panels to your needs, depending on your stimulation conditions.

In the spirit of fostering collaborative research at CU|AMC, antibody panels developed in the HIMSR will be shared with others on campus.

What antibodies are available through the HIMSR? Where should I look for protocols? A detailed list of antibodies and protocols are available on the HIMSR iLabs site. Go to “About HIMSR” on the the iLab website and scroll to “Links and Resources” at the bottom of the page for more information.