IHC Technical Resources

Protocols, optimization tips,
troubleshooting guides,
and more for IHC.

Troubleshooting guides

Troubleshooting guides

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handbooks for IHC, Western
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Sample collection and preparation play an important role in IHC as the antigen exhibition and location are largely depend on the quality of tissue sample.

  1. Cell Sample

    Adherent Cells

    • Cell Climbing: Grow adherent cells on multi-aperture culture plates with coverslip, culture vessels or chamber slide
    • Direct Cell Culture: Culture adherent cells directly on culture vessels or multi-aperture culture plates

    Non-Adherent Cell

    • Cell Smear: Adhere non-adherent cells on coverslip with chemical bond
    • Eccentric Cell Smears: Adhere non-adherent cells on culture vessels by cell micro-centrifuge
  2. Tissue Sample

    Tissue samples are typically taken from specimens of various sources: biopsy, surgery, animal model and autopsy. The first three types of specimens give fresh tissues while the last one (autopsy) is taken after an animal has died for two hours which is more or less a postmortem autolysis. As antigens may denature, disappear and diffuse, autopsy specimen should be fixated as soon as possible so as not to influence its label.

    Exercise caution when collecting, fixating and sectioning the samples

    • Use sharp knife and scissors to avoid extrusion damage
    • Cutter should be flat, small and thin (Normal size is 1.0 cm × 1.0 cm × 0.2 cm)
    • Eliminate fat tissue and calcification zone
    • Collect samples from live animals and fix samples immediately after wash
    • Choose diseased instead of necrotic region
    • Choose normal tissue as control if necessary
    • Make paraffin-embedded tissue or frozen tissue immediately after sectioning or store the tissues in liquid nitrogen container or refrigerator at -70°C