Immunostaining Techniques

Accurate interpretation of complex microscopic assays begins with excellent sample preparation and staining.

Our carefully executed SOPs and personal attention to each and every slide ensures your satisfaction.

Your team at Premier Laboratory has extensive experience developing assays for immuno staining and analysis.

Sample preparation, calibration, staining optimization, interpretation and reporting formats may be ordered in a standard or custom format.

We have an extensive in-house library of available antibodies, each of which has been validated and optimized for immunohistochemical or immunofluorescent staining.

Alternatively, your antibody and/or protocol can be incorporated.

The high-throughput automated staining systems at Premier Laboratory result in procedures that are executed with unmatched precision and consistency.

  • Advantages of Immuno and Molecular Staining

    Microscopic-based immuno staining techniques possess the advantage of being performed on intact tissue sections.

    Unlike techniques that rely on homogenized or disaggregated samples, the staining results from slide-based techniques are interpreted in context of tissue architecture, morphology and cell type.

    For complex tissues and heterogeneous samples (eg. breast carcinoma) using morphology as a gating parameter becomes a critical factor in confirming that your results are based on the specific cell population of interest, and therefore valid.

    For cell types that cannot be distinguished morphologically, antibodies can be used to locate or exclude an individual population.

    Multiparametric analysis, with double labeling or antibody cocktails, can be used to further elucidate specific populations.

  • Scalable Resources

    Scalable resources give you the capacity and precise tools you need for your immuno and molecular analysis.

    The extensive resources Premier Laboratory will put at your disposal include:

    • Board Certified Pathologists
    • A dedicated staff of highly qualified experienced scientists, technologists and technicians
    • The very latest in cutting-edge technology including high-throughput automated stainers and image analysis systems

  • Techniques

    • Immunohistochemistry (IHC)
    • Immunofluorescence (IFA)
    • New antibody protocol optimization & validation
    • Custom antibody panels
    • Double/multiple staining protocols
    • High-throughput screening with image analysis
    • Custom algorithm development (can incorporate automated pattern recognition)

  • Study Design

    Premier Laboratory provides you with consultative and expert advice on a wide array of routine and esoteric research protocols.

    Each study begins with a consultation to define your study objectives, allocate resources, and determine the optimal scientific approach.

    When combined these result in the design of the most cost effective, accelerated and efficacious study to meet your research goals.

    Our scalable product offering means you will achieve precisely the right combination of standard off-the-shelf and custom methodologies in the design of your research study.

    SOPs and systems are in place to support validated regulated (GLP) and non-regulated investigative protocols.

    Studies can be customized to include multiple antibodies, as well as to specify the level of interpretation.

    The scientific team at Premier Laboratory is always available to develop new protocols tailored to your exact needs.

  • Antibody Selection

    Select from Premier Laboratory’s extensive in-house library of high-quality antibodies that target known protein targets.

    Most antibodies have been characterized, validated and optimized for immunohistochemical and immunofluorescent staining, with standard protocols in place.

    Alternatively, your own antibody can be optimized for staining, or a Premier Laboratory scientist will assist you with the selection of the most appropriate commercially available antibody for your study.

    Premier Laboratory has extensive experience with the selection of single antibodies or antibody cocktails to provide co-localization information, as well as in double labeling.

    Your consulting scientist can assist you in selecting the right markers for differentiating the specific cell types needed for your research study.

  • Antibody Optimization and Antigen Retrieval

    Antibodies are optimized for strong signal-to-noise ratio, thereby eliminating confounding data that can result from nonspecific endogenous background staining.

    In addition, antigen retrieval techniques may be implemented to unmask binding sites that became unavailable during sample fixation and processing.

    These techniques allow for maximizing assay sensitivity and specificity, providing you with the most reliable data.

    When selecting an antibody from Premier Laboratory’s extensive in-house library, each has already been characterized, validated and optimized for immunohistochemical and immunofluorescent staining, with standard protocols in place.

  • Protocols

    Premier Laboratory protocols are developed with the right combination of standard off-the-shelf and custom methodologies to help you reach your research goals.

    Alternatively, we can implement your own study protocol.

    During your study design consultation, a Premier Laboratory’s scientist will work with you on the best choice of:

    • Antibodies
    • Sample processing technique
    • Selection of chromagen or fluorochrome and counterstain
    • Need for signal amplification
    • Staining procedure
    • Level of analysis/interpretation
    • Number of samples for significant statistical analysis

  • Sample Options

    • Quantification and analysis can be performed on your slides or on slides prepared for you by Premier Laboratory (from your frozen, fixed or embedded tissue)
    • Histologic sections, TMAs, smears or monolayers
    • Routine, special, immuno stained slides

  • Interpretation of Results

    Staining can be interpreted in a variety of ways from notation of presence/absence, to semi-quantitative (+1 to +4) analysis, to true quantification via high throughput digital image analysis.

    Knowledge of the expression patterns of a particular antibody, combined with evaluation of the correct area of biological interest, is key to generating significant results.

    Options for selecting the area(s) for quantification include:

    • Pathologist-based
    • Automated pattern recognition-driven
    • Standard algorithm
    • Custom algorithm

  • Quantification Option

    High throughput scanning and analysis with digital imaging, incorporating standard or customized algorithms, is available.

    Algorithms can incorporate automated pattern recognition programs.

    Advantages of digital image analysis include:

    • Quantitative rather than qualitative, with results amenable to statistical analysis
    • Repetitive tasks are scored with both objectivity and consistency
    • Measurements are precise with enhanced sensitivity
    • Rare event detection can be performed
    • Quantitative-based analysis – measure, count and classify
    • Complex analytical questions can be answered
    • Turnaround time is decreased
    • Generates a permanent data record
    • Analysis and documentation of subtle cellular changes not discernible to the human eye
    • Execution of high volume analysis

  • Digital Slide Option

    Premier Laboratory’s state-of-the-art high-throughput automated scanning service is ideal for the creation of your digital slide library.

    Your digital library will be robust, flexible and confidential.

    • 24-hour secure Internet enabled international access for remote viewing and analysis
    • Long-term image archiving
    • Database management (images with linked sample data)
    • Photomicrograph documentation
    • Coordinating images and associated data for publication
    • Importing select fields into text/presentation files (eg. Word or PowerPoint doc)
    • Image annotation and editing
    • Preparation for future image analysis
    • View and compare multiple slides side-by-side
    • Coordinating data from multiple projects
    • Generating permanent visual record of transient stains (eg. immunofluorescence)
    • Sharing images for review/consultation

  • Report Options

    Your report structure is determined during study design, and is based on your need for validated regulated (GLP) and non-regulated investigative results.

    Premier Laboratory’s flexible report format allows for customization depending upon the level of interpretation you require.

    Technical reports on methods, both for staining and analysis validation, are available.

    Complete statistical analysis packages may be incorporated.