Is there a bubble in the pore wall of the prefabricated receptor block purchased that will affect subsequent IHC results?

Incomplete pore edges increase the risk of flaking, especially during high-temperature antigen repair. It is recommended to use positively charged or silicided anti-slides, which can be appropriately lengthened to 60-65 degrees overnight or 70 degrees for 1 hour, but not too high to cause brittle tissue. Repair fluid boiling too much will cause the edges to tilt, it is recommended to turn down the fire or use a low microwave. For white tablets with gaps in the perforation, quantification of key markers or intergroup comparisons are not recommended and can be reserved for pre-test or pre-diluted antibodies.

At the time of receipt, it is recommended to check the hole wall bubbles at low temperature with a stereoscope or magnifying glass and take photos to record the batch. If there are many bubbles, dense or close to the hole wall, the supplier should be contacted for replacement; if it is only a small superficial bubble, the surface layer can be repaired 0.5-1.0mm before viewing. There is no problem with storing the wax block at 4 degrees, but after taking it out, it needs to be reheated to room temperature and then sliced to avoid new microcracks caused by thermal expansion and contraction. It can be sealed, protected from light and dust, and does not need to be placed in the refrigerator repeatedly.

Whether small bubbles near the orifice wall affect the IHC depends on whether the bubbles open to the edges of the tissue core. If the bubbles after slicing are only small vacuoles inside the wax block, the tissue core is still intact, and the effect on staining is usually limited; however, when the antigen is repaired, the liquid may penetrate along the microcracks, causing local high background or non-specific staining. It is recommended to cut 10 sheets in succession, confirm the integrity of the pore edge under the microscope, and then use a few of them as a negative control for the target antibody to observe whether there are any abnormal edges.

Bubbles generally do not expand significantly due to 4 degrees preservation, but if they are frequently taken from 4 degrees to room temperature, condensed water may form on the surface, which may affect the expansion of the section. If the bubbles are only on the surface of the pore wall, they can be eliminated or reduced after trimming; if the pore edge is still incomplete after slicing, IHC may cause abnormal edge staining due to reagent infiltration even if there is no obvious shedding. It is recommended to avoid the obvious area of bubbles when slicing, or to number each white sheet and record the cavity status, and exclude suspicious holes during analysis.