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Flow Cytometry

Accurately immunophenotype cells of interest

Comprehensively Quantify Therapeutic Effects through High Throughput Flow Cytometry Analysis of Various Cell Types

Gain new insight into your drug mechanism of action and PD through robust and in-depth immunophenotyping. Maximize immunotherapeutic benefit, limit unwanted toxicities, and increase your chances of preclinical/clinical success through comprehensively characterizing cells of interest. Use our high quality flow cytometry data to better understand:

  • The immune make-up of relevant organs such as tumor, blood, spleen, and lymph nodes
  • Therapeutic effect on the frequencies and functionality of cell subsets
  • Individual specimen variations and heterogeneity of response

Characterize Complex Immune-Phenotypes and Rare Cells of Interest with our Well-Established Flow Cytometry Platform

  • Characterize cells in high throughput with limited sample volume enabled by Miltenyi gentleMACS™ Dissociators, state-of-the-art equipment including BD FACSCalibur™, BD Accuri™ C6, NovoCyte, Beckman Gallios, BD LSRFortessa™ and BD LSRFortessa X-20 (up to 17 colors) with 96-well plate HTS attachment
  • Overcome tumor complexity with our pre-optimized list of over 20 protocols for tumor tissue dissociation and parent gating strategies
  • Ensure accurate staining, processing, and gating with our stringent controls and manual compensation
  • Eliminate human error through central data processing and analysis facilitated by data transfer software
  • Gain expert guidance from our dedicated FACS team to help determine the right dissociation protocols, panel configurations, gating strategies, etc. for your study
  • Modify reports to fit evolving needs with dynamic and informative reporting packages including a variety of raw data, statistical analyses, observations, actionable recommendations, data visualization, etc.
  • Easily and securely review high quality data and reports through our cloud portal

Our standard flow cytometry panels have been validated in multiple oncology and inflammation in vivo models to provide basic to comprehensive analysis of lymphoid and myeloid cell populations. Models analyzed include syngeneics, tumor homograft models, human target expressing (Humanized genetically modified mouse models / Humanized Target Tumor Cells), humanized, GvHD, and peritonitis models.

Advanced High-Parameter Analysis: Spectral Flow Cytometry

Unlock unprecedented depth in cellular analysis with Crown Bioscience's state-of-the-art Spectral Flow Cytometry services. Moving beyond conventional flow cytometry, spectral flow utilizes the entire emission spectrum of fluorophores, rather than relying on discrete bandpass filters. This revolutionary approach allows for the simultaneous detection and analysis of a significantly higher number of markers (typically 20-40+ parameters), providing a truly comprehensive view of complex biological systems.

Cytek® Northern Lights™ System (China location) Cytek® Northern Lights™ System (US SD location) Cytek® Aurora System (UK location)

Up to 3 lasers- blue, red, violet

35 fluorescence channels: 16V-14B-8R

Max up to 25 Color Panel

Up to 5 lasers- ultra violet, blue, yellow-green, red, violet

64 fluorescence channels: 16UV-16V-14B-10YG-8R

 

Up to 3 lasers- blue, red, violet

35 fluorescence channels: 16V-14B-8R

Max up to 25 Color Panel

 

Key Advantages for Your Research:

  • Unrivaled Parameter Capacity: Characterize intricate cellular populations and rare cell types with unparalleled resolution, essential for dissecting heterogeneous samples like tumor biopsies or complex immune repertoires.
  • Enhanced Data Quality: Minimize spectral overlap (spillover) and improve signal resolution, leading to cleaner, more accurate data for even dimly expressed markers.
  • Fluorophore Flexibility: Utilize a broader range of fluorophores, including those with highly overlapping emission spectra, optimizing panel design and expanding experimental possibilities.
  • Maximized Sample Insight: Extract more information from precious clinical or research samples, reducing the need for multiple assays and preserving valuable material.
  • Streamlined Analysis: Advanced spectral unmixing algorithms simplify data interpretation, making high-dimensional data more accessible and actionable.

Applications for Cancer Researchers:

Leverage spectral flow cytometry to:

  • Deeply Phenotype the Tumor Microenvironment (TME): Precisely characterize immune cell subsets (T-cells, B-cells, NK cells, myeloid cells, etc.) and their activation states within tumors, peripheral blood, and lymph nodes.
  • Accelerate Biomarker Discovery & Validation: Identify novel cell-surface and intracellular biomarkers for patient stratification, prediction of therapy response, and disease progression, by correlating complex phenotypic profiles with clinical outcomes.
  • Elucidate Drug Mechanisms of Action (MOA): Gain granular insights into how novel therapeutics impact target cells, immune populations, and resistance mechanisms.
  • Monitor Immunotherapy Responses: Track subtle changes in immune cell composition and function over time in patients receiving immunotherapies, providing crucial insights into treatment efficacy and potential adverse events.
  • Analyze Rare Cell Populations: Confidently detect and characterize rare tumor-infiltrating lymphocytes (TILs) or circulating tumor cells (CTCs) with high specificity and sensitivity.

Customized panels are also available, which undergo thorough validation prior to running your samples. Contact Us for more information.

See our standard panels and gating strategy below.

25-color Mouse Comprehensive Immunoprofiling Panel

N

Specificity

Purpose

Gating strategy

1

L/D

Viability

singlets>L/D-

2

CD45

Total leukocytes

L/D-CD45+

3

CD19

B cells

L/D-CD45+CD19+

4

CD27

B Memory

L/D-CD45+CD19+IgD-CD27+

5

IgD

B Naïve

L/D-CD45+CD19+IgD+CD27-

6

CD3e

Total T cells

L/D-CD45+CD19-CD3+

7

CD4

Helper T cells

L/D-CD45+CD19-CD3+CD4+

8

CD8

Cytotoxic T cells

L/D-CD45+CD19-CD3+CD8+

9

FoxP3

Regulatory T cells

L/D-CD45+CD19-CD3+CD4+FoxP3+

10

CD44

Naïve/Effector/Memory T

L/D-CD45+CD19-CD3+CD4+ or CD8+ CD44hiCD62L-(effector)

11

CD62L

Naïve/Effector/Memory T

L/D-CD45+CD19-CD3+CD4-or CD8+ CD44hiCD62L+(central memory), CD44lowCD62Lhi(naive)

12

CD335

NKT

L/D-CD45+CD19-CD3+CD335+

NK

L/D-CD45+CD19-CD3-CD335+

13

PD-1

Check point

L/D-CD45+CD19-CD3+CD4+ or CD8+PD-1

14

CD11b

Myeloid cells

L/D-CD45+CD19-CD3-CD335-CD11b+

15

Ly6G

Neutrophils/gMDSC

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G+

16

Ly6C

Monocytes/MDSC

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-Ly6Chi

17

F4/80

Macrophages

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-CD170-Ly6C-F4/80+

18

IA/IE/MHC II

M1

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-CD170-Ly6C-F4/80+IA/IE+

19

CD206

M2

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-CD170-Ly6C-F4/80+CD206+

20

PD-L1

Check point

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-CD170-Ly6C-F4/80+PD-L1+; L/D-CD45-SSChigh PD-L1+

21

CD11C

DCs

L/D-CD45+CD19-CD3-CD335-F4/80-Ly6G-CD11Cint Siglec H-CD11C+IA/IE+

22

Siglec H

pDC

L/D-CD45+CD19-CD3-CD335-F4/80-Ly6G-CD11Cint Siglec H+

23

XCR1

cDC1

L/D-CD45+CD19-CD3-CD335-F4/80-Ly6G-CD11Cint Siglec H-CD11C+IA/IE+CD11b-XCR1+

cDC2

L/D-CD45+CD19-CD3-CD335-F4/80-Ly6G-CD11Cint Siglec H-CD11C+IA/IE+CD11b+XCR1-

24

FcεRIα

Basophils

L/D-CD45+CD19-CD3-CD335-CD11C-IA/IE-FcεR1+

25

Siglec-F/CD170

Eosinophils

L/D-CD45+CD19-CD3-CD335-CD11b+Ly6G-CD170+

25-color Mouse Comprehensive Immunoprofiling Panel (TIL)

24-color Human Comprehensive Immunoprofiling Panel

N

Specificity

Purpose

Gating strategy

1

L/D

Viability

singlets>L/D-

2

CD45

Total leukocytes

L/D-CD45+

3

CD19

B cells

L/D-CD45+CD19+

4

IgD

Naïve B

L/D-CD45+CD19+IgD+

5

CD3

Total T cells

L/D-CD45+CD19-CD3+

6

CD4

Helper T cells

L/D-CD45+CD19-CD3+CD4+

7

CD8

Cytotoxic T cells

L/D-CD45+CD19-CD3+CD8+

8

TCR gd

Pan γδ T cell

L/D-CD45+CD19-CD3+TCRgd+

9

CD127 (IL-7Ra)

Regulatory T cells

L/D-CD45+CD19-CD3+CD4+CD25+CD127low/-

10

CD25

11

CD45RA

Naïve/Memory/Effector/TEMRA T cells

L/D-CD45+CD19-CD3+CD4+/CD8+CCR7+CD45RA+(TN), CCR7+CD45RA-(TCM)

12

CD197 (CCR7)

Naïve/Memory/Effector/TEMRA T cells

L/D-CD45+CD19-CD3+CD4+/CD8+CCR7-CD45RA-(TEM), CCR7-CD45RA+(TEMRA)

13

CD56

NKT

L/D-CD45+CD19-CD3+CD56+

NK

L/D-CD45+CD19-CD3-CD33-CD14-CD33-CD14-CD56highCD16-(CD56bright NK), CD56+CD16+(CD56dim NK)

14

CD11b

MDSC

L/D-CD45+CD19-CD3-CD56-CD123-HLADR-CD11b+CD33+

15

CD33

mMSDCs

L/D-CD45+CD19-CD3-CD56-CD123-HLADR-CD11b+CD33+CD14+CD15-

16

CD15

pMSDCs

L/D-CD45+CD19-CD3-CD56-CD123-HLADR-CD11b+CD33+CD14-CD15+

17

CD14

Classical Monocyte

L/D-CD45+CD19-CD3-CD56-CD123-HLA-DR+CD14+CD16-

Intermediate Monocyte

L/D-CD45+CD19-CD3-CD56-CD123-HLA-DR+CD14++CD16+

Non-classical Monocyte

L/D-CD45+CD19-CD3-CD56-CD123-HLA-DR+CD14+CD16++

18

CD16

Granulocytes

L/D-CD45+SSChighCD19-

Neutrophils

L/D-CD45+SSChighCD19-CD16highCD66b+

19

CD66b

Eosinophils

L/D-CD45+SSChighCD19-CD16-CD66b+

20

HLA-DR

DCs

L/D-CD45+CD19-CD3-CD33-CD14-CD33-CD14-CD16-CD56-HLADR+

21

CD11C

mDCs

L/D-CD45+CD19-CD3-CD33-CD14-CD33-CD14-CD16-CD56-HLADR+CD11C+

22

CD1c (BDCA-1)

cDC1

L/D-CD45+CD19-CD3-CD33-CD14-CD33-CD14-CD16-CD56-HLADR+CD11C+CD1C-

cDC2

L/D-CD45+CD19-CD3-CD33-CD14-CD33-CD14-CD16-CD56-HLADR+CD11C+CD1C+

23

CD123 (IL-3Ra)

Basophils

L/D-CD45+CD19-CD3-CD56-CD123+CD11C+HLA-DR-

24

CD163

CD163 in Monocytes; M2 like

CD163 Overlay in Monocytes

24-color Human Comprehensive Immunoprofiling Panel (Blood)

Investigate Mechanism of Action with Functional Assays

Characterize the biological responses of immune cells through flow cytometry based functional assays including receptor occupancy, apoptosis, and proliferation assays.

Discuss Your Flow Cytometry Project