COVID-19 Information

27-color ZE5 and YETI (710 Rubin)

The BioRad ZE5 was formerly known as the Propel Labs YETI.

Yeti

The ZE5 Cell Analyzer is a 27-color flow cytometer.  For more information on the capabilities of the ZE5, click here.  There are two ZE5 Cell Analyzers in the DartLab. Please contact DartLab staff to arrange for training if you would like to begin using these instruments.

 

Data Acquisition Software

Intuitive Everest software provides automated fluorescence compensation, a fluorochrome selector panel, and a runlist design wizard. Use this software on your computer to pre-configure the instrument settings, staining panels, and samples.  Integrated training modules, and the ability to analyze files while acquiring saves time and streamlines your workflow.  Everest software is PC-specific and is available at no charge from DartLab.

Optical Bench

Each graph depicts a single laser and shows the band width for each detector.  Be aware of inter-laser spectral overlap in detectors with similar band widths.

Filter sets for Yeti lasers

We have designed this template to download and enter fluors when designing a staining panel:

Yeti staining panel template

Data Analysis

"The high-throughput nature of flow cytometry, combined with the increasing capacity to measure more cell parameters at once, is generating massive and high-dimensional datasets on a routine daily basis. These data can no longer be adequately analysed using the classical, mostly manual, analysis techniques and therefore require the development of novel computational techniques, as well as their adoption by the broad community. Computational flow cytometry is a new discipline that straddles the fields of immunology and computational biology and provides a set of tools to analyze, visualize and interpret large amounts of cell data in a more automated and unbiased way." Saeys et al., 2016.

We recommend FlowJo V.10 for high-dimmensional data analysis. Please consult with us for guidance on data analysis strategies and use of plugins.

Available Fluorophores

UV Laser Fluors

BUV395

AlexaFluor350 (AF350), DAPI, Hoechst-Blue, Indo-hi

BUV496, Indo-lo

BUV661

BUV737, BUV 805, Hoechst-red

Do not use BUV563

Yeti UV laser filters

Violet laser fluors

BV421, Cascade Blue

BV450, Pacific Blue

BV510, AMCyan, Pacific Orange, Cascade Yellow

BV605

BV650

BV711

BV786

Filters for 405 nm laser

Blue laser fluors

Brilliant Blue 515, FITC, AF488, eGFP, eYFP

PE, PE-Dazzle, PE-CF594, PE-Texas Red (more useful off yellow laser)

PerCP-Cy5.5 but do not use PerCP (this is an excellent dump PMT)

PE-Cy7 (more useful off yellow laser)

Yeti blue laser fluors

Yellow laser fluors

PE

dTomato, DsRed

PE-Dazzle, PE-CF594, PE-Texas red

mPlum

PE-Cy5, PE-AF647

PE-Cy5.5

PE-Cy7

Yeti yellow laser fluors

Red laser fluors

APC, AF647, Cy5

APC-R700, AF680, AF700, DRAQ5, Cy5.5

APC-Fire750, APC-Cy7, APC-H7, AF750

AF790

Yeti red laser fluors

References

Start here with this excellent slide deck by Jonathan Irish.

Methods for discovery and characterization of cell subsets in high dimensional mass cytometry data (Diggins et al., 2015)

Characterizing cell subsets using marker enrichment modeling (Diggins et al., 2017)

A beginner's guide to analyzing and visualizing mass cytometry data (Kimball et al., 2018)