Immunohistochemistry (IHC)
Unveiling the intricate details of tissue samples through antibody-based detection, highlighting the distribution and localization of specific proteins.
Immunohistochemistry and Immunofluorescence
Immunohistochemistry (IHC) is a powerful laboratory technique used to detect specific proteins or antigens in cells of a tissue section. By using antibodies that bind to these proteins, IHC helps scientists and pathologists visualize the distribution and localization of specific molecular markers within a tissue sample. This technique is crucial in both research and clinical diagnostics, especially for identifying different types of cancer, infections, and other diseases.
Basic Principles of Immunohistochemistry
- Antigen-Antibody Binding: The core of IHC is based on the specific binding between an antigen (a protein or molecule present in the tissue) and an antibody (a protein designed to bind specifically to that antigen). When an antibody binds to its target antigen in a tissue sample, it can be detected using various methods that make the bound complex visible under a microscope.
- Sample Preparation: Tissue samples are typically fixed, often in formaldehyde, to preserve their structure and then embedded in a medium like paraffin wax. Thin slices of the embedded tissue are placed on slides for staining.
- Staining Methods: The binding of antibodies to their target antigens is not visible without a secondary detection system. Two main types of staining methods are used to visualize these bindings: immunofluorescence and chromogenic detection.
Staining Platform – BOND RX
The Leica Bond RX autostainer is a fully automated staining platform used for various histological and molecular pathology techniques.
Below is a summary of the main applications that can be realized on the Leica Bond RX autostainer:
Immunohistochemistry (IHC)
Overview: IHC is a technique used to detect specific antigens in tissue sections using antibodies. Applications: IHC on the Bond RX is commonly used to study the distribution and localization of biomarkers in tissue samples.
Immunofluorescence (IF)
Overview: fluorescently labeled antibodies detect antigens in tissue sections.
Applications: IF is frequently used in research for co-localization studies
In Situ Hybridization (ISH)
Overview: ISH is a method used to detect specific nucleic acid sequences (DNA or RNA) within tissue sections. Both, chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH) can be performed.
Applications: ISH is valuable for detecting gene expression, chromosomal abnormalities and many more.
RNAscope
Overview: RNAscope is an advanced ISH technology designed to detect RNA molecules with high specificity and sensitivity.
Applications: RNAscope is particularly useful for single-cell RNA expression analysis
Multiplex Immunohistochemistry (mIHC) and Multiplex Immunofluorescence (mIF)
Overview: Multiplex staining allows for the simultaneous detection of multiple targets within a single tissue section. Both, chromogenic multiplexing (mIHC) and fluorescent multiplexing (mIF) protocols can be performed.
Applications: These methods are used to study complex cellular interactions, tumor microenvironment, immune profiling, and biomarker discovery in research.
Custom Protocols
Overview: We can implement custom staining protocols tailored to specific research needs. We can modify existing protocols or create new ones, adjusting parameters such as incubation times, temperatures, and reagent volumes.
Applications: Custom protocols are invaluable in experimental research settings where novel or modified techniques are required for unique research questions can be adjusted to our customer’s needs.
Opal Kit: We have extensive experience with the Akoya Biosciences Opal Detection Kit and can realize stainings up to 6 colors.
Digital Pathology and Imaging Technologies
With the advent of digital pathology, traditional methods like IHC and ISH are enhanced by advanced imaging and analysis technologies, allowing for more accurate, reproducible, and high-throughput pathology workflows.
Image Acquisition- PhenoImager
We can provide a state-of-the-art digital pathology platform designed for multiplexed immunofluorescence imaging and analysis using the the PhenoImager from Akoya Biosciences. It integrates automated slide scanning, multispectral imaging, and digital pathology, making it ideal for both research and clinical applications.
Functionality:
- Multiplexing: The PhenoImager allows for the simultaneous detection of multiple markers within a single tissue section, which is critical for studying complex cellular environments, such as tumor microenvironments.
- Multispectral Imaging: Images across different wavelengths, separating overlapping fluorescent signals can be captured, thereby increasing the sensitivity and specificity of detection.
- Automated Scanning: The system automatically scans entire slides at high resolution, providing consistent and high-quality images of the stained tissues.
Image Analysis – HALO Image Analysis Software
HALO software, developed by Indica Labs, is a comprehensive image analysis platform used for quantitative analysis of tissue images. It supports both brightfield and fluorescence imaging, making it versatile for various pathology applications.
Capabilities:
- Quantitative Analysis: HALO provides robust tools for quantifying IHC and ISH staining, including cell counting, measurement of staining intensity, and spatial analysis of markers.
- Machine Learning and AI: The software incorporates machine learning algorithms that can be trained to recognize and quantify specific cell types or staining patterns, improving the accuracy and efficiency of image analysis.
- Batch Processing: We can analyze large datasets simultaneously, streamlining workflows and allowing for high-throughput analysis using HALO software.
- Applications: Widely used in research and clinical diagnostics for quantitative pathology, biomarker validation, and understanding disease mechanisms.