Immunohistochemistry of immune tissues

Aim:

With this assay, the presence and location of proteins can be demonstrated in tissue sections.

The aim of these immunohistochemical procedures is to investigate

• the localization and tissue distribution of target molecules,
• the changes in the expression level/concentration of target molecules in the tissue by fluorescent microscopy (3 channels).

Test system:

In vitro, the following tissues can be investigated:

• Formalin fixed paraffin-embedded (FFPE) tissues
• Frozen sections
• Human, NHP, other animal tissues (healthy or specific disease panel, full tissue array from commercial source)

Design:

The evaluation method can be qualitative, semi-quantitative and quantitative (morphometrical analysis, cell counting).

Establishment of staining procedure (selection of antibody – primary and detection system, positive and negative control tissues, optimization of fixation, antibody concentrations, incubation time, blocking method)

Validation of the staining procedure (if necessary) by testing the following parameters:

• Specificity
• Sensitivity
• Repeatability
• Dose-dependence

Outcome:

Depending on the type of evaluation:

• Qualitative – negative or positive
• Semi-quantitative – scoring
• Quantitative – number of cells, area of positivity per visual field