Tissue microarray immunohistochemistry (IHC) double labeling

Service Description:

Immunohistochemistry (IHC) or immunocytochemistry (ICC) is a technique that uses labeled specific antibodies to perform in situ qualitative, localization, or quantitative studies of the distribution and content of certain chemical components in tissue sections or cell specimens at the tissue and cellular level. This project is a tissue microarray immunohistochemistry double-labeling experiment. Tissue microarray IHC can arrange dozens to hundreds of small tissues neatly on a single slide for staining, ensuring consistent experimental conditions for tissue samples on the chip, saving time and reagents, and facilitating unified analysis after scanning. Compared with single-slide staining, both time and cost can be reduced. The disadvantage is that the selected tissue area is relatively small and cannot comprehensively reflect the expression profile of individual samples. The tissue microarray IHC workflow is basically the same as that of conventional tissue IHC, but because the samples are relatively precious, the operational procedures are more stringent.

Main Applications:

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 with the aid of microscopic imaging and magnification. In pathology, IHC is often used for tumor diagnosis and differential diagnosis, and it can also be used to further refine tumor classification.

Detection Principle:

Based on antigen-antibody reactions (i.e., specific antigen-antibody binding) and chemical color development principles, exogenous antibodies (or antigens) with labels are introduced and anchored to the corresponding antigens (or antibodies) in tissue or cell specimens. The label then produces a color reaction to show 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 - elution - antigen retrieval - inactivation of endogenous peroxidase and biotin - serum blocking - primary antibody incubation - secondary antibody incubation - TY reagent - red color development - hematoxylin counterstaining - dehydration and clearing - mounting - microscopic examination.

Sample Requirements and Transport Conditions:

1. Provide properly fixed and qualified samples. After tissue removal, immediately fix with 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 cell爬片 must be fully fixed. Paraffin sections must be fully deparaffinized before the experiment.

3. Provide accurate antibody information.

Notes:

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