Cytek Investor Day Presentation Deck
Application of FSP in Immunology
Development of a 43 color panel for the characterization of
conventional and unconventional T-cell subsets, B cells, NK cells,
monocytes, dendritic cells, and innate lymphoid cells using
spectral flow cytometry
Fairooz Sahir, Jericha Miles Mateo, Martin Steinhoff, Kodappully Sivaraman Siveen X
First published: 18 December 2020 | https://doi.org/10.1002/cyto.a.24288 | Citations: 2
Funding information: Hamad Medical Corporation, Grant/Award Numbers: MRC-03-19-039, IRGC-04-51-
17-151, IRGC-03-NI-17-071
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Abstract
Although many flow cytometers can analyze 30-50 parameters, it is still challenging to
develop a 40+ color panel for the phenotyping of immune cells using fluorochrome
conjugated antibodies due to limitations in the availability of spectrally unique
fluorochromes that can be excited by the commonly used laser lines (UV, Violet, Blue,
Green/Yellow-green, and Red). Spectral flowcytometry is capable of differentiating
fluorochromes with significant overlap in the emission spectra, enabling the use of
spectrally similar fluorochrome pairs such as Brilliant Blue 515 and FITC in a single panel.
We have developed a 43 color panel to characterize most of the immune subsets within
the peripheral immune system, including conventional T cells, unconventional T cells
such as invariant natural killer T cells (INKT), Gamma delta (yo) T-cell subsets (TCR V82,
TCR VY9) and mucosal-associated invariant T cells (MAIT), B-cell subsets, natural killer (NK)
cells, plasmacytoid dendritic cells, dendritic cell subsets, hematopoietic progenitor cells,
basophils, and innate lymphoid cell (ILC) subsets (CD117, CRTH2). The panel includes
surface markers to analyze activation (CD38, HLA-DR, ICOS/CD278), differentiation
(CD45RA, CD27, CD28, CD57), expression of cytokine and chemokine receptors (CD25,
CD127, CCR10, CCR6, CCR4, CXCR3, CXCR5, CRTH2/CD294), and co-inhibitory molecules
and exhaustion (PD-1, CD223/LAG-3, TIGIT), which enables a deep characterization of
PBMCs from peripheral blood. Cells were analyzed on a 5-laser Cytek Aurora and data
analysis was done using Flowjo. This panel can help to make a thorough interpretation of
immune system, specifically when specimen quantity is low. The panel has not been
completely optimized but would rather act as a guide toward the development of a
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Scientists developed a 43 color panel to characterize most of the immune
subsets within the peripheral immune system, including
conventional T cells
unconventional T cells
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B-cell subsets
natural killer (NK) cells
dendritic cells
A Selection of Application Areas for FSP
CYTEK
hematopoietic progenitor cells
basophils
Innate lymphoid cell
Cells were analyzed on a 5-laser Cytek Aurora and data analysis was done
using Flow Jo.
This panel can help to make a thorough interpretation of immune system,
specifically when specimen quantity is low.
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