Spectral Flow Cytometry Capabilities

Spectral unmixing for deeper immunoprofiling
Intuitive panel design for customizing panels
Autofluorescence extraction for improved sensitivity and resolution

How does spectral flow cytometry work?

Spectral flow cytometry offers unique optical collection and analytical capabilities enabling the use of a wide range of fluorochrome combinations without reconfiguring the system for each application, resulting in exceptional sensitivity with simple and intuitive operations. Full spectrum signatures are generated for each fluorochrome bound to cellular markers that are used to identify different cell types. Different signatures present in a multicolor sample are then unmixed to allow the use of fluorochromes with close emission peaks in the same sample. This expansion in fluorochrome options and panel size potential, along with the vacuum-powered fluidics system, makes the Cytek® Aurora spectral flow cytometry particularly suitable for studies where unique biomarker signatures are sought or sample availability is limited.

In a conventional flow cytometer, pacific blue and brilliant violet 421 (BV421) cannot be used together as the spectral signatures are too similar. In spectral flow cytometry, pacific blue and BV421 have enough distinctions (a complexity index of 0.78) in their spectral signatures to be used in combination.

Advantages of spectral flow cytometry

More detection

5 lasers with 64 fluorescence detectors

Greater Flexibility

Use of a wide array of new fluorochrome combinations without the need to reconfigure optical filters

Outstanding Sensitivity

Unmixing of fluorochrome with close emission peaks
Use of autofluorescence extraction to improve data clarity

More colors

Detection of up to 40 different cell types/targets in the same multicolor sample

High Value System

Larger datasets generated from smaller sample sizes

Advanced technology with full analytical compatibility and familiar workflows

Spectral flow cytometry offers several advantages over conventional flow cytometry and at the same time is amenable to complex analyses of multiparameter flow cytometric data such as the t-Distributed Stochastic Neighbor Embedding (tSNE) analysis that allows for an unbiased analysis of all markers in the panel to help discern phenotypic changes within cell subsets that would otherwise go undiscovered. Spectral flow cytometry has all the capabilities of conventional flow cytometry, follows the same workflow and shares the same requirements, while expanding its functionalities.

4T1-Luc2-1A4 tSNE analysis Plotting traditional gating populations over the tSNE map allows the visualization of shifts in the expression of any panel markers (population shifts) between treatment groups

Advanced technology with full analytical compatibility and familiar workflows

Big insights from small samples

Our expert flow cytometry team can perform deep immunophenotyping investigation of your in vivo study to capture shifts that occur in the immune response triggered by test agent treatment. Below is a list of our standard spectral panels. These panels have the capacity for additional markers to be added and completely custom panels can be developed for your assay needs.

Connect with a Representative

Provides immunophenotypic analysis of 19 distinct myeloid subsets

Antibody/Dye	Description
CD45	Pan-hematopoietic cell marker
CD45 (mouse)	Mouse immune cell exclusion
CD3	Pan-T cell marker
CD4	CD4+ T cell marker
CD8	CD8+ T cell marker
FoxP3	Regulatory T cell marker
CD56	Natural killer/Natural killer T cell marker
CD16	Differentiation marker
CD19	B cell marker
CD20	B cell marker
CD11b	Pan-myeloid lineage marker
CD68	Macrophage marker
HLA-DR	M1 macrophage marker
CD163	M2 macrophage marker
CD14	Monocyte marker
CD33	Myeloid delineation
CD84	MDSC marker
CD11c	Dendritic cell marker
BDCA3 (CD141)	DC1 subset marker
BDCA1 (CD1c)	DC2 subset marker
CCD123	pDC marker/Basophil marker
CD66b	Neutrophil marker
CD15	Neutrophil marker
CD86	Maturation marker
Viability Dye	Dead cell exclusion

Provides absolute counts, quantitation of 13 lymphoid and myeloid subsets, memory T cell measurements and analysis of 12 functional markers to profile T cell, dendritic cell, macrophage and MDSC phenotypes

Panel 1: Antibody/Dye	Description
CD45	Pan-hematopoietic cell marker
CD3	Pan-T cell marker
CD4	CD4+ T cell marker
CD8	CD8+ T cell marker
FoxP3	Regulatory T cell marker
CD25	Regulatory T cell marker
CD44	Activation/Memory marker
CD62L	Naïve T cell/Memory marker
PD-1	Activation/Exhaustion marker
CD69	Activation marker
LAG-3	Activation/Exhaustion marker
TIM-3	Exhaustion marker
ICOS	Activation marker
Granzyme B	Cytotoxicity marker
CKi-67	Proliferation marker
CD49b/CD335	Natural killer/Natural killer T cell marker
CD19	B cell marker
CD11b	Pan-myeloid lineage marker
Ly-6G	Granulocytic-MDSC/ Neutrophil marker
Ly-6C	Monocytic-MDSC/Monocyte marker
CD11c	Dendritic cell marker
CD24	Dendritic cell delineation
F4/80	Pan-macrophage marker
MHC Class II	M1 macrophage & dendritic cell marker
CD206	M2 macrophage marker
CD103	DC1 subset marker
XCR1	Cross presenting/Migratory dendritic cell marker
CD80	Maturation marker
CD86	Maturation marker
MARG1	Myeloid functional marker
iNOS	Myeloid functional marker
PD-L1	Immunosuppression
Viability Dye	Dead cell exclusion

Panel 2: Antibody/Dye	Description
Counting Fluorosphere Beads	Cell enumeration reagent
CD45	Pan-hematopoietic cell marker
Viability Dye	Dead cell exclusion

Provides deep phenotypic analysis into T cell, NK and B cell subsets, combining 11 activation and exhaustion markers. Included is the delineation of naïve stem cell memory, central memory, effector memory, terminal effector and tissue resident memory (Trm) subsets.

Antibody/Dye	Description
CD45	Pan-hematopoietic cell marker
CD3	Pan-T cell marker
CD4	CD4+ T cell marker
CD8	CD8+ T cell marker
TCRgd	Gamma Delta T cell marker
FoxP3	Regulatory T cell marker
CD25	Activation/regulatory T cell marker
CD69	Activation marker
LAG3	Activation/Exhaustion marker
CD27	Activation marker
ICOS	Activation marker
TIM3	Exhaustion marker
PD-1	Activation/Exhaustion marker
TIGIT	Exhaustion marker
TCF7	Exhaustion marker
Granzyme B	Cytotoxicity marker
CXCR5	T cell recruitment
CXCR3	T cell recruitment
CD56	Natural killer/Natural killer T cell marker
CD16	Natural killer/Natural killer T cell marker
CD19	B cell marker
CD20	B cell marker
Ki-67	Proliferation marker
CD45RA	Memory T cell delineation
CD45RO	Memory T cell delineation
CCR7	Memory T cell delineation
CD62L	Memory T cell delineation
CD95	Memory T cell delineation
CD103	Memory T cell delineation
Viability Dye	Dead cell exclusion