The rockefeller university

Panel Design Step 1: Marker Expression and Co-expression – 8:01(opens in new window)

Panel Design Step 2: Fluorochrome Selection – 10:27(opens in new window)

Panel Design Step 3: Spillover-Spreading Error -7:30(opens in new window)

Panel Design Step 4: Designing a Panel – 10:47(opens in new window)

Panel Design Step 5: Evaluate Panel Performance – 7:11(opens in new window)

Webinar 20211130 – Autofluorescence Characterization Using Full Spectrum Profiling™ (FSP™) by Kate Pilkington – 41:32(opens in new window)

Webinar 20210325 – Just Because You Don’t See It Doesn’t Mean It Isn’t There: Improved Assay Resolution with Autofluorescence Extraction by Alexis Gonzalez – 44:50(opens in new window)

Reagent / Antibody titration:

The Power of Reagent Titration in Flow Cytometry. Diana LB, Alberta P, Jesus G, Lily MP, Maria CJ. Cells. 2024 Oct 11;13(20):1677(opens in new window)

OMIP-069: (opens in new window)Forty-Color Full Spectrum Flow Cytometry Panel for Deep Immunophenotyping of Major Cell Subsets in Human Peripheral Blood. Park LM, Lannigan J, Jaimes MC. Cytometry A. 2020 Oct;97(10):1044-1051(opens in new window)

OMIP-069 Version 2: (opens in new window)Update to the 40-color full Spectrum flow cytometry panel for deep immunophenotyping of major cell subsets in human peripheral blood. Park LM, Lannigan J, Low Q, Jaimes MC, Bonilla DL. Cytometry A. 2024 Nov; 105(11):791-799(opens in new window)

OMIP-102: (opens in new window)50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells. Andrew JK, Peter LM, Aaron JT, Martin P, Florian M. Cytometry A. 2024 Jun;105(6):430-436(opens in new window)

OMIP-109: 45-color full spectrum flow cytometry panel for deep immunophenotyping of the major lineages present in human peripheral blood mononuclear cells with emphasis on the T cell memory compartment. Lily MP, Joanne L, Quentin L, Maria CJ, Diana LB. Cytometry A. 2024 Nov;105(11):807-815(opens in new window)

Panel Optimization for High-Dimensional Immunophenotyping Assays Using Full-Spectrum Flow Cytometry. Laura Ferrer-Font, Sam J Small, etc. Current Protocols. 2021 Sep;1(9):e222(opens in new window)

Panel Optimization for High-Dimensional Immunophenotyping Assays Using Full-Spectrum Flow Cytometry. Laura Ferrer-Font, Sam J Small, etc. Current Protocols. 2021 Sep;1(9):e222 – PDF

Additional Resources:

Plan Your Assay

      Sign Up

• • Free Cytek Cloud(opens in new window) account to access all spectral panel design tools
  • Aurora User Forum(opens in new window) (and look in the Application sub-forum to find Normalization spreadsheets, configuration charts, and more!)

      Panel Design

• • Watch a video series on panel design in Cytek® SpectroLearn™(opens in new window)
  • Publication about Panel Design Best Practices(opens in new window)
  • Webinar: Building Panels for Flow Cytometry: Key Steps for Success(opens in new window)
  • OMIP-069(opens in new window) Cytek’s 40 Color Panel publication – A comprehensive resource for designing and optimizing high parameter panels on Cytek Instruments (Don’t forget to grab the supplement too!)

      Reference Controls

• • Review best practices in the Reference Control video in Cytek® SpectroLearn™(opens in new window)
  • List of tandem and non-tandem dyes (attached)

      Titration

• • Webinar on Debunking Antibody Titration Myths(opens in new window)
  • Antibody Titration Protocol(opens in new window)
  • Blog article by Laura Johnston about the vital importance of properly titrating viability dyes(opens in new window)

Run Your Assay

Check out the collection of video tutorials about SpectroFlo(opens in new window) if you want more in depth detail about specific features, including:

• • System Startup & QC(opens in new window)
  • Create and Unmix an Experiment(opens in new window)
  • Plate loaders: AMS loader(opens in new window) and ASL loader(opens in new window) (newer version)
  • Note that we release frequent software updates so please refer to your TAS or applicationsupport@cytekbio.com(opens in new window) for the most up-to-date advice

Gating reference controls(opens in new window) – another great blog article by Laura Johnston about setting up gates in the unmixing wizard

Webinar on instrument standardization(opens in new window)

 Evaluate Your Assay

• • Publication about Panel Optimization Best Practices(opens in new window)
  • Cytek White Paper about Evaluating Panel Performance(opens in new window)

Autofluorescence

For more detailed information on AF, check out these webinars:

• • The Power of Autofluorescence Extraction Using Full Spectrum Profiling™(opens in new window)
  • Autofluorescence Characterization Using Full Spectrum Profiling™ (FSP™)(opens in new window)
  • Full Spectrum Profiling™ (FSP™) Enhances Resolution of Samples with Complex Autofluorescence(opens in new window)