Immunohistochemical demonstration of leucocyte differentiation antigens on paraffin sections using a modified AMeX (ModAMeX) method

The AMeX method (cold Acetone fixation with subsequent Methyl benzoate and Xylene treatment and routine paraffin embedding) has been recently revived for simultaneous preservation of morphology of cells and their antigens. We propose a modification of this method (ModAMeX), with the use of proteolytic enzyme inhibitors and low temperature paraffin wax embedding, which results in better preservation of a large number of leucocyte differentiation antigens and diagnostic morphologic detail. T-cell antigens (CD1, CD2, CD3, CD7 & CD8), B-cell antigens (CD22), macrophage associated antigens (CD11c, CD14 and others), activation antigens (CD25 and others), as well as some other antigens of diagnostic interest (CD10) were found to be preserved with a staining intensity equal to that of sections of fresh frozen tissue. Although the staining intensity of other T-cell antigens (CD4 & CD5), B-cell antigens (CD19, CD21 & CD37), activation antigens (Ki-1) and nuclear proliferation antigen (Ki-67) was slightly weaker as compared with frozen sections, this could be corrected by increasing the monoclonal antibody concentration. Staining for heavy and light chains of immunoglobulins was minor, sometimes compromised due to persistence of background staining as a result of extracellular immunoglobulins. The ModAMeX method has the advantages of simplicity, low cost and the possibility of exchange of tissue material between laboratories.