In Vivo Immunophenotyping via Flow Cytometry

Flow cytometry is the tool of choice to determine the immunomodulatory action of new immunotherapeutics during animal testing. Our large suite of cell line-derived and mouse-derived syngeneic tumor models allows the testing of immunotherapeutics with various modes of action.

Features of our flow cytometry service:

We employ a designated team of scientists with rich experience in flow cytometry and cell sorting applications
A suite of state-of-the-art flow cytometers and cell sorters covering all flow cytometry needs
Four ready-to-go staining panels are available for immune phenotyping with multiplexing of up to 20 immune cell markers
Panel customization and de novo panel developments are an option

Reach out today to discuss your project plans. We will be happy to work with you to ensure your specific research questions are being addressed to the best extent possible.

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Instrumentation – Flow cytometers and Cell sorters

Flow cytometer
Flow cytometer Instrument	Laser	Marker	Comments
Sony SP6800ZE Spectral Analyzer	3	20	full spectrum analyzer uses spectral unmixing instead of conventional compensation

Cell sorter
Cell sorter Instrument	Laser	Marker	Comments
BD FACS Aria Fusion	4	18	Sort 4 cell populations simultaneously
BD FACS Aria III	5	21	Sort 4 cell populations simultaneously
Beckman Coulter MoFlo Astrios EQ	4	21	Sort 6 cell populations simultaneously

Our panels

We offer four staining panels for immediate immune profiling of tumor samples. Custom-tailored panels can be established upon request to detect the immune cell population of your interest.

Four staining panels for immediate immune profiling
Marker	All-In-One Panel	NK / T Cell Panel	T Cell Activation Panel	DC Panel
B220	x			x
CD107a			x
CD11b	x	x
CD11c	x			x
CD122		x
CD172a				x
CD19		x		x
CD206	x
CD25	x		x
CD3	x	x	x	x
CD335	x
CD4	x	x	x
CD40				optional
CD44		x
CD45	x	x	x
CD49b	x	x
CD62L		x
CD69			x
CD8	x	x	x
CD80				optional
CD86				optional
F4-80	x			x
FoxP3	x	x	x
GrzB			x
IFNg			x
Ki67		x
Lag-3			optional
Ly6C	x
Ly6G	x
MHCII	x			x
PD1		x
Siglec H				x
Tim3			optional
XCR1				x

Gating strategy

All-In-One Flow Panel

Detected cell populations:

CD4+ T cells

CD8+ T cells

Treg cells

NK cells

B cells

M-MDSCs

PMN-MDCSs

DCs

M1 / M2 Macrophages

Neutrophils/ Granulocytes

NK / T Cell Panel

Detected cell populations:	Memory status:	Activation markers:
CD4+ T cells	CD62L	PD-1
CD8+ T cells	CD44	Ki67
Treg cells		CD122
NK cells
NKT cells

T Cell Activation Panel

Detected cell populations:	Activation markers:	Optional markers:
CD4+ T cells	Granzyme B	Lag3
CD8+ T cells	IFN-gamma	Tim3
Treg cells	CD107a (lamp 1)
	CD69

DC Panel

Detected cell populations:		Additional markers:
T cells		PD-L1
B cells		CD86
Macrophages		CD40 (optional)
cDC1		CD80 (optional)
cDC2		CD86 (optional)
pDC

Flow Cytometry for Immunophenotyping Setup

Clinical data indicate that only some patients respond to treatment with immune-modulating drugs. Therefore, it is essential to consider the immune response during preclinical drug development. Flow cytometry is widely used for the analysis of immune cell populations in tumors as a means to describe the immune response.

Panel: A comprehensive analysis of numerous immune cell populations in one tumor is a challenge. We use established, ready-to-use staining panels that allow for the analysis of various immune cell populations. In addition, we can custom-tailor our existing panels or establish new antibody panels to match our customer’s research needs.

Method: Primary tumor material is disrupted, erythrocytes removed, and up to 3×10⁶ obtained single cells/well are dispensed. The single cells are stained with marker-specific fluorescent antibodies and analyzed by flow cytometry.

Report: All data of the mouse study, including tumor growth and flow cytometry data, will be presented in a comprehensive report, custom-tailored to the client study, and written by a Ph.D.-level technical writer. The analysis enables you to draw conclusions directly from the study outcome and use the data for the filing of official documents.

Add-ons: Further immunophenotyping methods are available at Reaction Biology to investigate the cytokine & chemokine concentrations in the tumor via multiplex immunoassays, to localize tumor-infiltrating immune cells via IHC, or to test the immune cell function via in vitro assays.

In Vivo Immunophenotyping via Flow Cytometry

Instrumentation – Flow cytometers and Cell sorters

Our panels

Gating strategy

Flow Cytometry for Immunophenotyping Setup

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