Our fully automated multispectral immunofluorescent (mIF) slide stainer and scanner allows rapid, high-throughput processing of your FFPE samples from tissue staining to data analysis and interpretation in under six weeks, all while maintaining tissue morphological integrity. Spectral unmixing allows synchronous assessment of up to eight biomarkers on each tissue section, enabling assessment of the cell cycle, tumor microenvironment, or any other biomarkers pertinent to your specific study. Finally, our analysis software delivers spatial mapping and quantification of your phenotypes of interest. Our in-house bioinformaticians can also assist with data interpretation and banking in our image repository.

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Limitations of traditional immunofluorescence
  • Limited to 3-4 markers
  • Autofluorescent background
  • Spectral bleed through
  • Low signal-to-noise ratio
  • Low confidence in quantitative analysis of data
Advantages of multispectral unmixing
  • Multiplex greater than 4 markers
  • Removal of background autofluorescence
  • Removal of spectral bleed through
  • High signal-to-noise ratio
  • Reliable and quantitative analysis of data

Panels

Hu Cell Cycle Slides
Example cell cycle panel staining of a genitourinary cancer tissue microarray. Click for larger
Hu Immune Cell Slides
Example immune panel staining of a gynecological cancer tissue microarray. Click for larger

Standard, optimized cell cycle and immune panels are available for both human and mouse samples. Custom biomarker panels are also available. ATISR requires any biomarkers to have a commercially available IHC-validated antibody for panel inclusion. Panels can consist of up to eight biomarkers plus DAPI.

Immune 1

  • CD4
  • FOXP3
  • CD163
  • PD1
  • CD8
  • AE1AE3

Immune 2

  • TIM3
  • CD4
  • Gal-3
  • CD8
  • LAG3
  • AE1AE3

Immune 3

  • CD3
  • FOXP3
  • CD8
  • GrB1
  • PD1
  • AE1AE3

Immune 4

  • PD-L1
  • CD3
  • OX40
  • CD33
  • CD163
  • PD1
  • CD8
  • AE1AE3

Immune 5

  • B2AR
  • CD3
  • CD8
  • CD15
  • CD33
  • LAG3
  • PD1
  • AE1AE3

Immune 6

  • CD3
  • CD8
  • CD163
  • OX40
  • PD-1
  • PD-L1
  • FOXP3
  • AE1AE3

MDSC

  • PD-L1
  • CD206
  • CD68
  • CD33
  • CD8
  • CD15
  • CD11c
  • AE1AE3

Neutrophil

  • LAIR-1
  • B7-H4
  • CD4
  • CD8
  • CD14
  • CD15
  • PD-L1
  • AE1AE3

Exhaustion 1

  • CD80
  • CD86
  • CD4
  • CD8
  • LAG3
  • TIM3
  • CTLA4
  • AE1AE3

Exhaustion 2

  • LAG3
  • CD45
  • PD1
  • CD8
  • TIM3
  • CTLA4
  • K167
  • AE1AE3

Cell Cycle 1

  • MCM2
  • pRB
  • RB
  • CCND1
  • CCNE
  • Ki67
  • pHH3
  • AE1AE3

Cell Cycle 2

  • MCM2
  • pRB
  • TAZ
  • CCND1
  • CCNE
  • CAMK1D
  • pHH3
  • AE1AE3

Cell Cycle 3

  • MCM2
  • pRB
  • CCND1
  • CCNE
  • CCNA
  • pHH3
  • AE1AE3

Cell Cycle 4

  • p21
  • p16
  • CCNE
  • CCND1
  • GMNN
  • CDK2
  • RB
  • AE1AE3

RB/pRB

  • MCM2
  • pRB
  • RB
  • c-CASP3
  • p21
  • pS6
  • pHH3
  • AE1AE3

pFAK

  • CD68
  • CD45
  • CD163
  • pFAK
  • HLA-DR
  • FAK (IFR8)
  • CD8
  • AE1AE3

BCL2

  • AR
  • BCL2
  • VIM
  • CD45
  • Ki67
  • AE1AE3

NE Lineage

  • AR
  • pRB
  • SYP
  • YAP
  • TAZ
  • Ki67

NE Lineage

  • pRB
  • CHG-A
  • pHH3
  • SYP
  • MASH-1
  • PanCK

Myeloid

  • CD11b
  • CD45
  • pHH3
  • CD8
  • LY6C
  • LY6G
  • PD1
  • PanCK

Immune

  • PD-L1
  • CD3
  • CD8
  • CD11c
  • CD163
  • FOXP3
  • Ki67
  • AE1AE3

GvHD

  • MUC2
  • LGALS3
  • LGR5
  • LYZ
  • VIL1
  • CHG-A

Lung Lineage

  • EGFP
  • CDH1
  • EGFRL858R
  • YAP
  • SYP
  • VIM
  • Ki67
  • PanCK

Workflow

Diagram of Akoya’s spectral unmixing and removal of tissue autofluorescence
Diagram of Akoya’s spectral unmixing and removal of tissue autofluorescence.
Hoyt CC. Multiplex Immunofluorescence and Multispectral Imaging: Forming the Basis of a Clinical Test Platform for Immuno-Oncology. Front Mol Biosci. 2021 Jun 2;8:674747. doi: 10.3389/fmolb.2021.674747. PMID: 34150850; PMCID: PMC8208831.
inForm software analysis workflow
inForm software analysis workflow.
Akoya Biosciences. Spatial Phenotyping in Clinical and Translational Research. eBook.

Technology

Leica BOND RXm Fully Automated Stainer

Leica BOND RXm Fully Automated Stainer

This fully automated slide stainer allows us to optimize and run your multiplexed biomarker panels in a timely and highly reproducible manner.

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AKOYA PhenoImager HT

AKOYA PhenoImager HT

This automated slide scanner delivers high throughput images from both brightfield and multispectral fluorescent staining. It allows for unmixed annotated regions of interest for panels of eight biomarkers and full slide unmixed images for panels of six biomarkers.

The accompanying inForm and phenoptr software enables rapid turnaround of spatial analysis and biomarker quantification of your samples.

 

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