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  • Introduction
  • CD2/CD4/CD8

Triple staining techniques is a recent developement still in immunohistochemistry. The use of these techniques is becoming more common in departments with it building on the advantages seen in dual staining technqiues.

Triple staining techniques, as the name implies is used to test for the presence of 3 different cell types in the same tissue. This can be used to differentiate in cases where diagnosis can be difficult to acheive, for example in complex lymphoma cases

In the reaction shown below, triple staining takes advantage of the ability of CD4, CD8 and CD20 to stain up different cell populations. Using different processes of staining with Nitroblue tetrazolium used for CD8, DAB for CD4 and Fuschin for CD20. This as shown below allows a direct comparison between the cells in the same tissue.

Picture 1 – Triple staining use in Lymph node


This is one of the most useful triple staining techniques in ICC as it can be used to distinguish between T and B cell lymphomas as well as determination of the cell origin of NK cells or helper T cells. This is a area that is growing in importance and is highly likely to expand to other laboratories. The only drawback of using this technique is that it is time consuming, which is a limiting factor but it still has a promising future