Integrated histology for quantitative digital pathology

Tissue-based biomarker investigation is a key step in pre-clinical / clinical drug development as it can serve either identification of novel therapeutic target, assessment of a surrogate marker of drug efficacy as well as prediction of a candidate compound benefit. To support novel therapies development at both pre-clinical and clinical level, we offer expert histology services relying on automated and standardized stainings capabilities spanning from monoplex to 7-plex marker panels and using either conventional chemistry or fluorescence revelation systems (immunohistochemistry/immunohistofluorescence).

Many types of samples – either tumor or normal tissue, originating from human or pre-clinical models – can be adressed using our platform. Multispectral images acquired on our PhenoImager HT (Akoya Biosciences) are processed through image analysis to provide quantitative data including phenotyping, marker expression, as well as geographical distribution of cells within the samples. In addition, bespoke services can also be offered to specifically meet our clients’ needs.

Illustrative data

Mature TLS revealed by multiplexed-IHF in a liposarcoma
Image field of mature TLS revealed by multiplexed IHF in a primary dedifferentiated liposarcoma: The mature stage of TLS was evaluated using an immunofluorescence approach multiplexing CD4, CD8, CD20, CD21 and CD23 markers. Results are representative of 540 tumors analyzed (discovery cohort, n = 328; validation cohort A, n = 131; validation cohort B, n = 81).

Image field of mature TLS revealed by multiplexed IHF in a primary dedifferentiated liposarcoma: ...

Multiplexed-IHC imaging in human lung adenocarcinomas
Multiplexed immunohistochemistry imaging for immune contexture analysis in human lung adenocarcinomas
(A) A human FFPE section was automatically stained on a Ventana discovery platform using a combination of antibodies targeting Cytokeratin 7 (CK7, for tumor cells, pink), CD163 (for tumor-associated macrophages, green), and CD8 (for CD8 T cells, brown). Image was originally acquired through a multispectral imaging system (Vectra® Polaris™, Akoya Biosciences), and then processed through inForm® analysis software (Akoya Biosciences) and machine learning approach for tissue segmentation (tumor and stroma regions) and then for cell segmentation & phenotyping. 
(B) Immune cell density (CD8+ and CD163+) quantification for each patient (n=80) within the different tissue regions highlighted a highest immune cell density within the stroma compared to the tumor. In addition, this analysis revealed an expected heterogeneous infiltration profile that could underlie sensitivity to therapies.
(C) In addition to cell density analysis, proximity of immune cells (CD8+ or CD163+) from tumor cells (CK7 positive) has been calculated for each patient (n=80) and, again, demonstrated a patient-dependent pattern.

Multiplexed immunohistochemistry imaging for immune contexture analysis in human lung adenocarcinomas ...

Tumor samples analyzed by multiplexed- IHF
Analysis of baseline tumor samples through multiplexed immunohistofluorescence    Cholangiocarcinoma patients tumor samples were analyzed by immunohistofluorescence (Ventana Discovery Ultra, Roche) multiplexing CD8 (Green) / CD163 (Yellow) / PD1 (Red) / PDL1 (Orange) / IDO1 (Cyan) / CK19 (White) / DAPI (Blue) markers. Following their acquisition (Vectra Polaris, Akoya Biosciences), multispectral images were processed for each marker deconvolution. This analyse revealed that responder patients were characterized by a higher expression of PDL1 as well as IDO1.

Analysis of baseline tumor samples through multiplexed immunohistofluorescence    ...

Key advantages of tissue-based imaging

  • Exploration at multiple scales, from cell-to-cell interactions to the macroscopic tissue architecture.
  • Well-suited for simultaneous detection of multiple markers to identify specific cell subsets, document an immune response or quantify biomarkers of interest.
  • Automated and robust workflow from sample processing to multiplexed image acquisition and quantification.
  

Automated tissue slide imaging

Platform

Discovery XT (Ventana / Roche Diagnostics)

Staining type

  • Immunohistochemistry (IHC)
  • Immunohistofluorescence (IHF)

Sample format

  • Paraffin embedded tissues (FFPE sections)
  • Frozen tissues (Cryo-sections)
  • Tissue Micro Arrays (TMAs)

Multiplexed quantitative pathology imaging

Platform

PhenoImager HT (Akoya Biosciences)

Sample format

Brightfield and fluorescence whole-slide scanning

Multispectral imaging

Up to 9 colors multiplexing

Readouts

Integrated tissue analysis software packages for  marker/biomarker quantification and spatial analysis