T Cell Assay Kit

8-Plex for Human PBMC Samples
AbKT-1003-10RXN

VistaPlex™ Assay Kits for CellScape offer a ready to use, reliable and customizable solution to fast-track precise spatial multiplexing.

Designed to provide a convenient, flexible, and modular analysis of immune cells, the T-Cell Assay kit enables quantification of 8 phenotypic biomarkers that define T-cell populations and subpopulations in human PBMC samples. Validation was performed on the CellScape platform on 4 blood donors in duplicate, in addition to individual antibody validation. The T Cell Assay Kit contains primary antibodies and antibody diluent and is optimized for ease of use and throughput.

PBMC sample showing 8 biomarkers quantified across immune cells. The biomarkers are used to phenotype T-cell populations, such as those shown here with defining markers displayed.
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Biomarkers identified and quantified:
CD3CD25CD39CD8
CD4CD27CD45RACD45
Cell population characterized and quantified:
LeukocytesT-CellsT-cytotoxicT-helper
T-regulatoryNaive CD4+Central memory CD4+Effector CD4+
Effector memory CD4+Naive CD8+Central memory CD8+Effector CD8+
Effector memory CD8+

Details & Data

Gating Strategy

Hierarchical gating strategies define cell phenotypes. From image data, each cell is captured by automated segmentation, and the biomarker expression is quantified using HDR imaging to measure fluorescence intensity for every cell. Standard FCS files are generated to enable the identification of cell phenotypes via classic flow cytometry-like hierarchical gating.

Representative dot plots demonstrating a gating strategy to identify T-cell subtypes.

Example Gating Strategy

Cell PopulationParent GateGating Strategy
LeukocytesAllCD45+
T cellsCD45+ LeukocytesCD45+ CD3+
T cytotoxicT cellsCD45+ CD3+ CD4- CD8+
T helperT cellsCD45+ CD3+ CD4+ CD8-
T regulatoryT helperCD45+ CD3+ CD4+ CD8- CD25+ CD39+
Naive CD4+T helperCD45+ CD3+ CD4+ CD8- CD27+ CD45RA+
Central memory CD4+T helperCD45+ CD3+ CD4+ CD8- CD27+ CD45RA-
Effector CD4+T helperCD45+ CD3+ CD4+ CD8- CD27- CD45RA+
Effector memory CD4+T helperCD45+ CD3+ CD4+ CD8- CD27- CD45RA-
Naive CD8+T cytotoxicCD45+ CD3+ CD4- CD8+ CD27+ CD45RA+
Central memory CD8+T cytotoxicCD45+ CD3+ CD4- CD8+ CD27+ CD45RA-
Effector CD8+T cytotoxicCD45+ CD3+ CD4- CD8+ CD27- CD45RA+
Effector memory CD8+T cytotoxicCD45+ CD3+ CD4- CD8+ CD27- CD45RA-

Reliable

The CellScape platform captures biological variability, while maintaining technical reproducibility. For the T-Cell Assay Kit, technical replicates were tested on 4 different donors. Fresh PMBC samples were used to confirm intra-assay and inter-assay reproducibility. To ensure high quality data can be obtained on a variety of sample types, the assay kit was successfully tested on samples that were biobanked on CellScape™ Chips for 6-months and cryopreserved samples.

Ready to Use and Customizable

The T-Cell Assay Kit has been optimized to maximize throughput by minimizing the number of cycles required for analysis. One open imaging channel for filter set FS395 in each cycle allows for easy customization. Additional cycles may be added for further customization of your high-plex assay.

T-Cell Assay Kit Stain Plan

CycleAntibodyCorresponding Filter SetVol. AntibodyDiluent VolumeIncubation Time
1CD3FSPerCP10μL260μL5 min
CD25FS56010μL
CD39FS48810μL
CD8FS42110μL
2CD4FSPerCP10μL260μL5 min
CD27FS56010μL
CD45RAFS48810μL
CD45FS42110μL

T-Cell Assay Kit Documentation

Download the Validation Report for the T-Cell Assay Kit.

Store all kit contents at 4°C, protected from light.

Additional materials required: CellScape Cell Chips (PRSM-CHP-CELL-010) and Wash Buffer (PRSM-BUF-WASH-1L).

Download Validation Report
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