Invicro’s Advanced Pathology Services (APS) team provides end-to-end solutions to support translational and clinical research by providing a combination of routine and unique tissue testing services. As part of the REALM IDx, Inc. group, our pathology service laboratories support projects spanning preclinical biomarker discovery research to clinical trials and development of companion diagnostics (CDx) products. Our scientific, operational and regulatory expertise enables us to deliver insights needed to take lead drug candidates to the next level.
As an early-phase and clinical service provider, we offer routine and novel pathology services, including core histology, specialty stains, multiplex Immunohistochemistry (IHC), multiplex immunofluorescence (IF), Advanced Cell Diagnostics (ACD) RNAScope®, BaseScope™ and miRNAScope™ ISH with the option to perform select assays in a CAP-accredited, CLIA-certified laboratory. With laboratory locations in Boston, MA, Irvine, CA, and Japan, we are able to support projects spanning all phases of drug discovery and development.
Invicro’s Advanced Pathology Services multidisciplinary team provides end-to-end discovery, translational research and clinical trial pathology services for pharmaceutical and biotechnology partners. We support studies across many therapeutic areas, including oncology, immuno-oncology, neurology, immunology and systemic diseases. Our unparalleled scientific expertise coupled with our operational capabilities allows us to support a variety of project types.
For all projects processed in our laboratory, our dedicated scientific and project management teams collaborate closely with sponsors to design studies that will deliver the insights needed to support research efforts at all stages of discovery and development. When partnering with Invicro, we ensure both sides are aligned on expectations and provide the highest level of transparency expected from a contract research partner.
- Receptor binding assays
- Cryofluorescence tomography (CFT)
- 2D and 3D quantitative autoradiography (ARG)
- Whole-body autoradiography (ARG)
- Micro-autoradiography (MARG)
- Image analysis
Translational & Clinical Research
- Assay design, development and validation
- Routine & specialty stains
- Immunohistochemistry (IHC) / Immunofluorescence (IF)
- Quanticell™ High sensitivity IHC
- RNAScope®, BaseScope™ and miRNAScope™ ISH
- Whole Slide Imaging (WSI)
- Image analysis and reporting
- Pathologist consultation and immunostaining interpretation
- Non-GLP Tissue Cross-Reactivity (TCR) assays
- 510(k) tissue-based submission tests for medical devices
- Regulatory filing support
- Companion Diagnostic (CDx) development for IHC and RNAScope® ISH
Download our translational tissue biomarker services flyer here!
Quanticell™ High Sensitivity Immunohistochemistry (IHC)
Quanticell is advanced and highly sensitive immunohistochemistry-based technology that accurately detects and quantitates specific target antigens at the molecular level supporting drug discovery and development. This quantitative immunohistochemistry assay uses novel fluorescent phosphor-integrated dots (PIDs) and provides superior sensitivity and uniformity compared to conventional immunofluorescent imaging and immunohistochemical methods. PIDs are organic fluorophore assembly-conjugated nanoparticles with unique properties including: uniform size, high intensity fluorescent signal, high photostability (densely packed fluorophores networked via chemical binding), high sensitivity, and broad dynamic range. At Invicro, our image analysis team uses the latest artificial Intelligence-based capabilities to analyze tissues stained with this innovative technology.
To overcome specific detection-related challenges, this assay offers higher sensitivity compared to common chromogenic and fluorescent-based detection methods. This service enables sponsors to:
- Visualize and quantitate low expressing targets and immune cells
- Evaluate and measure drug distribution alongside the drug target
- Monitor and quantify drug pharmacodynamic effects in the tissue context
- Assess and predict drug response in patient tissues
- Versatile: Track and measure localization of therapeutics: monoclonal antibodies, antibody-drug conjugates, bispecifics and recombinant proteins
- Sensitive: Detect and quantify low expressing targets while maintaining tissue morphology
- Predictive: Assess drug mechanism of action to build more reliable therapy response models
- Accurate: Reduce detection of false negatives
- Flexible: Compatible with typical FFPE IHC antibodies and multiplex staining methods in a single tissue section
- Guo Z, Tada H, Kitamura N, et al. Automated Quantification of Extranuclear ERα using Phosphor-integrated Dots for Predicting Endocrine Therapy Resistance in HR+/HER2- Breast Cancer. Cancers (Basel). 2019;11(4):526. Published 2019 Apr 12. doi:10.3390/cancers11040526
- Fujisawa T, Tsuta K, Yanagimoto H, et al. Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD-L1 antibodies. Mol Clin Oncol. 2019;10(3):391-396. doi:10.3892/mco.2019.1801
- Inamura K, Shigematsu Y, Ninomiya H, et al. CSF1R-Expressing Tumor-Associated Macrophages, Smoking and Survival in Lung Adenocarcinoma: Analyses Using Quantitative Phosphor-Integrated Dot Staining. Cancers (Basel). 2018;10(8):252. Published 2018 Jul 31. doi:10.3390/cancers10080252
- Yamaki S, Yanagimoto H, Tsuta K, Ryota H, Kon M. PD-L1 expression in pancreatic ductal adenocarcinoma is a poor prognostic factor in patients with high CD8+ tumor-infiltrating lymphocytes: highly sensitive detection using phosphor-integrated dot staining. Int J Clin Oncol. 2017;22(4):726–733. doi:10.1007/s10147-017-1112-3
- Gonda K, Watanabe M, Tada H, et al. Quantitative diagnostic imaging of cancer tissues by using phosphor-integrated dots with ultra-high brightness. Sci Rep. 2017;7(1):7509. Published 2017 Aug 8. doi:10.1038/s41598-017-06534-z
- Optimization of an ultrasensitive, quantitative immunoassay for detection of CD20 in Non-Hodgkin’s Lymphoma (NHL) FFPE samples. Goel et al. (2020). The Society for Immunotherapy for immunotherapy of Cancer (SITC).
- Quanticell™, a novel histopathology technology for visualization and quantitation of membrane PSMA demonstrated in cell lines and clinical tissue. Dimant et al. (2020). AACR.
- Examining “Her2 Low” patient populations for prediction to Her2-targeting antibody-drug conjugate response in Her2 negative patients using a novel high sensitivity immunohistochemistry assay. Krueger et al. (2020). AACR.
- Evaluating loss of STING/cGAS expression using a novel multiplex immunohistochemistry detection approach. Goel et al. (2019). The Society for Immunotherapy of Cancer (SITC).
- Quantitative measurement of human epidermal growth factor receptor-2 (HER2) protein expression in ‘classical’ and ‘non-classical’ FISH categories: a comparative study. Shen et al. (2018). USCAP.
- Prognostic value of nuclear and non-nuclear estrogen receptor expression by IHC with phosphor-integrated dots in hormone receptor-positive early breast cancer Guo et al. (2018). AACR.
Immunohistochemistry (IHC) is a powerful tissue-based method used both in research and clinical diagnostics to evaluate pathological conditions. IHC allows for the visualization of target antigens on the cellular and subcellular level with tissue morphology preserved in a wide-range of sample types, such as solid tumor biopsies, lymph nodes and other isolated tissues. In clinical practice and drug development, IHC is widely used as a companion diagnostic test to stratify patients and monitor the efficacy of therapeutic treatments. With rapid advancements made in the field of immunotherapy, IHC remains a valuable assay to support the development and approval of cutting-edge therapies.
Our laboratories possess the scientific, regulatory and operational expertise to develop and run immunohistochemistry (IHC) assays across multiple therapeutic areas. From early-stage discovery to exploratory endpoint testing and CDx development, our team is prepared to support all your project needs to expedite drug discovery and development efforts. We perform IHC on xenograft tumor models, tissue microarrays (TMA), and a diverse population of human and non-human tissue samples. It is our mission to provide biomarker insights needed to help you make smart decisions that will accelerate drug development efforts.
With multiple laboratory locations and CAP-accredited CLIA-certified sites, we offer comprehensive IHC services to support projects with varying levels of complexity. Our laboratories are equipped with the latest research and clinical tissue processing equipment from Leica, Agilent Dako, Biocare Medical and Ventana to support your specific project needs.
Core Service Capabilities
- Scientific consultation
- Tissue processing & embedding
- FFPE/fresh frozen serial sectioning
- Custom assay development
- CAP/CLIA assay validation
- Singleplex, multiplex IHC
- Multiplex IHC combined with RNAScope® ISH
- Whole slide imaging (WSI)
- Pathologist interpretation
- Non-GLP Tissue Cross-Reactivity (TCR) assays
- 510(k) tissue-based submission tests for medical devices
Multiplex Immunostain Examples
Immunofluorescence (IF) is a powerful approach to detect and visualize multiple (e.g., 4-plex, 8-plex) biomarkers while preserving tissue morphology to understand spatial relationships. Multiplex Immunofluorescence is an effective approach to examine the colocalization of infiltrating immune cells within the tissue microenvironment, which is particularly important in immuno-oncology drug development.
Developing multiplex immunofluorescent assays and analyzing the data is a challenging process that requires domain knowledge and expertise. Invicro’s team of scientific experts provide a consultative approach to study design, execution, analysis, and reporting. Our approach starts with understanding the question that needs to be answered followed by designing a study that will deliver new insights required to support translational research. We are a platform agnostic solutions provider and can develop new assays using a wide-range of staining chemistries, including tyramide signal amplification (TSA)-based Akoya Opal sequential staining, DNA barcoding-based Ultivue InSituPlex, Ventana multiplex panels, and others. All staining workflows can be developed on automated platforms, including the Leica Biosystems BOND RX or the Ventana DISCOVRY ULTRA.
Core Service Capabilities
- Scientific consultation
- Study design
- Tissue processing & embedding
- FFPE serial sectioning
- Antibody optimization (e.g., monoplex chromogenic/fluorescent optimization)
- Assay detection optimization
- Multiplex IF combined with RNAScope® ISH
- Multispectral imaging
- Image analysis
Digital Pathology & Image Analysis
Invicro’s dedicated image analysis team provide end-to-end digital pathology capabilities to support all drug research and development phases from biomarker discovery to clinical trials. By partnering with us, we provide research services with rapid turnaround times (TATs) to help uncover biological insights to advance your scientific research. We deploy state-of-the-art analysis workflows to identify phenotypes within the tissue context. In addition to providing combined tissue processing and analysis, we accept digitized slides for analysis only projects to provide clients access to tailored project solutions. To digitize slides for analysis applications, our laboratories are equipped with a broad portfolio of commercial, state-of-the-art systems and image analysis tools.
By combing cutting-edge technologies and software, including Visiopharm and other in-house custom developed tools to meet your specific needs, with our deep expertise in data analytics, we deliver solutions that support brightfield and multispectral fluorescent imaging techniques used to detect biomarkers and phenotype immune cells in the tumor microenvironment (TME). We help clients solve complex research studies by offering analysis solutions for all testing methods, including core histology, multiplex IHC/IF and Quanticell–Konica Minolta’s proprietary high sensitivity, quantitative IHC assay.
Core Analysis Services
- Tumor scoring
- Immune cell counting
- Multiplex stain quantification
- RNAScope® BaseScope™ and miRNA™ ISH analysis
- Multiplex IHC combined with ISH to visualize gene and protein targets
- Morphological analysis
- Custom application development/transfer
Digital Biomarker Solutions
Invicro has established a strategic alliance with Paige, a leader in clinical-grade artificial intelligence (AI) for diagnostics, drug discovery and biomarker development applications. With this partnership, Invicro now provides biopharmaceutical clients access to world-class digital pathology solutions across a range of disease indications. The computational pathology solutions support efforts spanning from preclinical discovery studies to clinical trials and diagnostic product commercialization.
Paige has produced AI solutions that generalize and phenotype tissue specimens without the need for labor-intensive and subjective annotation-based methods. In addition, access to critical data, such as genomics, treatment information, drug response, and other clinical assessments is available to augment and gather insights from clinical studies. Through this partnership, we address core challenges including:
- Lack of access to curated pathology datasets
- Need for expertise in managing clinical studies
- Access to regulatory expertise
Pharma Service Offerings
- Custom Biomarker Discovery Studies
- Multi-modality Studies
- Clinical Product Development & Commercialization
Core Histology & Specialty Stains
In addition to our routine immunohistochemistry and novel quantitative biomarker imaging solutions, Invicro provides multiple specialty stains to support your research efforts across multiple disease areas.
|Alcian Blue-PAS-hematoxylin stain||Alkaline Phosphatase stain|
|Aniline Blue stain||Bielschowsky’s silver stain|
|Goldners Trichrome stain||Gomori’s Trichrome stain|
|Luxol fast blue and PAS stain||Masson’s Trichrome stain|
|Methylene blue/basic fuchsin stain||Movat pentachrome stain|
|Periodic acid-methenamine-silver stain (PAM)||Picrosirius red/van gieson picrofuchsins stain|
|Safranin O/fast green stain||Sirius red stain|
|Toluidine blue Stain||TRAP stain|
|TUNEL stain||Von kossa stain|
Clinical Trial Services
In addition to our translational biomarker services, Invicro provides clinical pathology services to support clinical trial and companion diagnostic (CDx) development programs. Our CAP-accredited and CLIA-certified laboratory possess the operational experience to optimize and validate biomarker assays, including CAP-CLIA validations and analytical validations needed for FDA submissions. We have established processes to ensure our laboratory remains fully compliant with all regulatory requirements for clinical studies.
- Inspect sample quality and accession into LIMS system for sample chain of custody
- Fix, process, section FFPE or fresh frozen tissues
Custom Assay Development / Assay Transfer
- Source and optimize commercially available or sponsor’s proprietary antibodies, identify control tissues and/or cell lines, identify isotype controls, optimize antigen retrieval conditions, select commercial staining platform
Stain Tissue Sections
- Stain primary tissues/tissue microarrays (TMA) using pre-optimized assay conditions and controls
Whole Slide Imaging (WSI)
- Scan slides at 20X or 40X to digitize slide (generate svs files) for image transfer or image analysis
Interpretation and Scoring
- Board-certified pathologist (MD) review, including qualitative review, establishing cutoffs, and IHC scoring
- Segmentation, morphological assessment, cell counting, distribution analysis, multiplex stain analysis using in-house tools and of the shelf software, including Visiopharm and other custom developed tools
- Deliver final report to sponsor, including methodologies utilized, complete analytical study results, statistical analyses
- Collate pathology image data using iPACS®–Invicro’s flagship data management platform to streamline data accessibility
Cryofluorescence Tomography (CFT)
CFT is an imaging technique performed on whole frozen animals (e.g., rats, mice, etc.) or organs that provides white light anatomical and molecular fluorescence 3D images with micron scale resolution. CFT bridges the gap between fluorescence histology and bulk fluorescence imaging by providing high-resolution imaging of multiple fluorophores simultaneously in a range of sample sizes.
High resolution, large tissue images are captured using sequential section-image cycles. This allows a high-throughput imaging workflow for whole animals or multiple organs (e.g., rodent organ, whole rodent body, dog or monkey head) by embedding multiple samples per block. The 3D mapping of the fluorescent signal is done with anatomical white light and images can be viewed slice-by-slice as a flythrough of the white light and fluorescence images, or via 3D multiplane rendering.
We approach all projects with a data-driven mentality and understand that quality data is critical to solving scientific challenges. Our team is constantly innovating and developing efficient processes to extract meaningful and valuable quantitative information to drive the drug development process forward.
- Evaluate the biodistribution of biological therapies including: oncolytic viruses, monoclonal antibodies (mAb), antibody drug conjugates (ADC) and bispecific fusion proteins
- Track the biodistribution and transduction of gene therapies using different routes of administration
- Track and visualize CAR-T cells, natural killer (NK) cells, dendritic cells, macrophages and stem cells
- Visualize and characterize transgenic and reporter models
- High Sensitivity: Compared to BLI/FLI, CFT offers higher sensitivity to detect signal deep in tissue
- Superior Resolution: Provides highest resolution compared to traditional whole animal imaging modalities
- Increased Throughput: Process several mice or multiple dissected organs on the same block
- Robust: Reduce data variability by combining CFT with in vivo imaging and tissue microscopy from the same animal
- Simple: No fixation, perfusion, tissue clearing, or radiolabeling required to maintain complete anatomical context
- Evaluating delivered gene therapies in vivo to evaluation site of gene transduction
- Determining the Pharmacokinetics (PK) and Pharmacodynamics (PD) of intrathecally administered ASOs
- Detecting spread of metastatic lesions in preclinical animal models using CFT
- Visualizing the renal clearance of two fluorescent-based tracers
- Developing a forward model for CFT imaging simulation
- Mazur C, Powers B, Zasadny K, et al. Brain pharmacology of intrathecal antisense oligonucleotides revealed through multimodal imaging. JCI Insight. 2019;4(20):e129240. Published 2019 Oct 17. doi:10.1172/jci.insight.129240
- Wolf DA, Hesterman JY, Sullivan JM, et al. Dynamic dual-isotope molecular imaging elucidates principles for optimizing intrathecal drug delivery. JCI Insight. 2016;1(2):e85311. Published 2016 Feb 25. doi:10.1172/jci.insight.85311
- Macrophage tracking using multi-modality 3D imaging in xenografts. Dimant et al. (2020). AACR.
- Multi-modality imaging study to determine AAV biodistribution compared to AAV transduction in whole animals. Dimant et al. (2019). Gene Therapy for Neurological Disorders Summit.
- Cryo-fluorescence tomography as a new tool in 3D visualization of tumor heterogeneity, metastatic proliferation, and immuno-oncology. Brevard et al. (2018). AACR.
- 3D assessment of antibody distribution within clinical tumors using cryofluorescence tomography. Dimant et al. (2017). WMIC.
- Download the CFT service flyer here!
Autoradiography (ARG) is an imaging technique used to assess the distribution of radiotracers in whole animal (i.e., mice and rats), discrete organs, tissues and cells. Whole body ARG allows for the visualization of drug pharmacokinetics and drug penetration into specific targets during the lead optimization process of drug screening. This imaging modality provides quantitative outputs and can be combined with other high-resolution optical modalities (e.g., bright field for anatomical referencing). In addition, Microautoradiography (MARG) offers higher resolution (0.3 µm) for imaging soft tissue section to evaluate the distribution of radiotracers on the cellular and sub-cellular levels.
- Biodistribution and quantitation of radiotracers
- Build 3D models from 2D Autoradioluminograms to quantify drug distribution in whole organs or animals
- Radioligand binding studies for receptor characterization
- Quantitative whole body autoradiography (QWBA) to determine organ biodistribution
- Quantitative: Quantify multiple classes of small molecule drugs and biological therapeutics
- Sensitive: Detects nanocurie amounts of radiotracer introduced into biological systems
- High Resolution: Delivers highest resolution on the micron scale compared to traditional in vivo imaging modalities
- Medium Throughput: Process multiple whole animal and dissected organs on the same block
- IIovich O, Qutaish M, Hesterman JY, Orcutt K, Hoppin J, Polyak I, Seaman M, Abu-Yousif AO, Cvet D, Bradley DP. Dual-Isotope Cryoimaging Quantitative Autoradiography: Investigating Antibody-Drug Conjugate Distribution and Payload Delivery Through Imaging. J. Nucl Medicine. 2018;59(9): 1461. Published 2018 May 4. doi: 10.2967/jnumed.118.207753.
- Development of a medium-throughput method to screen the effect of test articles on mouse brain activity using 14C-2-deoxyglucose (14C-2DG) 3D autoradiography. Sullivan et al. (2018). NRM.
- Download the ARG service flyer here!