Tissue microarray immunohistochemistry (IHC)

Service Introduction:

Using labeled specific antibodies to qualitatively, spatially, or quantitatively analyze the distribution and content of certain chemical components in tissue sections or cell specimens in situ at the tissue and cellular level is called immunohistochemistry or immunocytochemistry. Tissue microarray immunohistochemistry allows dozens to hundreds of small tissue samples to be neatly arranged on a single slide for staining, keeping experimental conditions consistent across tissue samples on the chip, saving time and reagents, and facilitating unified analysis after scanning. Compared with staining individual slides, both time and cost can be reduced. The disadvantage is that the selected tissue area is relatively small and cannot fully reflect the expression status of individual samples. The workflow of tissue microarray immunohistochemistry is essentially the same as that of conventional tissue immunohistochemistry, but because the samples are relatively precious, the operating procedures are more stringent.

Main Applications:

By using the imaging and magnification of microscopy, immunohistochemistry can detect various antigenic substances (such as proteins, polypeptides, amino acids, polysaccharides, phospholipids, enzymes, hormones, pathogens, and receptors) at the cellular and subcellular levels. In pathology, immunohistochemistry is commonly used for tumor diagnosis and differential diagnosis, and can also be used for more detailed classification of tumors.

Detection Principle:

Based on the antigen-antibody reaction (i.e., specific binding between antigen and antibody) and chemical color development principles, labeled exogenous antibodies (or antigens) are introduced and anchored to the corresponding antigens (or antibodies) in tissue or cell specimens; the label produces a color reaction to visualize the target antigen (or antibody).

Experimental Workflow:

Deparaffinization and rehydration - antigen retrieval - inactivation of endogenous peroxidase and biotin - serum blocking - primary antibody incubation - secondary antibody incubation - DAB color development - hematoxylin counterstaining - dehydration and clearing - mounting - microscopic examination.

Sample Requirements and Transport Conditions:

1. Provide properly fixed samples. After tissue removal, immediately fix in an appropriate fixative; the volume of fixative is recommended to be more than 10-15 times the tissue volume. Fresh specimens should be fixed in appropriate containers for 24-48 h; large specimens should be fixed for 12 h and then cut open for further fixation. Specimens should be fixed at room temperature (around 24°C) or refrigerated (4°C); do not freeze. Fixed samples should be transported at room temperature.

2. Smears and coverslip cell preparations must be fully fixed, and paraffin sections must be thoroughly deparaffinized before the experiment.

3. Provide accurate antibody information.

Notes:

1. A preliminary experiment on a single sample should be performed first. The sample can be provided by the customer or prepared by our company. Charges are based on the number of sections (same cost as conventional IHC) to confirm antibody specificity and the optimal concentration.