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Updated: Aug 8, 2026
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Immunohistochemical analysis of Hodgkin's disease using microwave heating
C Charalambous1, N Singh, P G Isaacson
1University College, London Medical School, Department of Histopathology, London.
Insights
Microwave heating significantly enhances immunohistochemical staining for CD15 and CD30 antigens in Hodgkin
Area of Science:
- Histopathology
- Immunohistochemistry
- Oncology
Background:
- Hodgkin's disease diagnosis relies on identifying Reed-Sternberg cells.
- Immunohistochemistry is crucial for detecting CD15 and CD30 antigens in these cells.
- Optimizing staining techniques improves diagnostic accuracy.
Purpose of the Study:
- To evaluate the impact of microwave heating on CD15 and CD30 antigen staining.
- To compare microwave-assisted staining with conventional methods for Hodgkin's disease.
- To determine the optimal conditions for enhanced immunohistochemical detection.
Main Methods:
- Tissue sections from 20 Hodgkin's disease cases were used.
- Immunohistochemical staining for CD15 and CD30 was performed.
- Sections were treated with conventional methods, microwave heating, and a combination including an IgM bridge.
Main Results:
- Microwave heating substantially increased the number and intensity of CD15-stained cells.
- The combination of Leu-M1 antibody, IgM bridge, and microwave heating yielded optimal CD15 staining.
- Microwave heating dramatically enhanced CD30 staining and eliminated non-specific plasma cell staining.
Conclusions:
- Microwave heating significantly improves immunohistochemical staining for CD15 and CD30.
- This technique is highly recommended for Reed-Sternberg cell antigen detection in Hodgkin's disease.
- Enhanced staining improves diagnostic reliability in Hodgkin's lymphoma.
Aims:
To assess the effect of microwave heating on immunohistochemical staining of CD15 and CD30 antigens in Hodgkin's disease tissue samples.
Methods:
Formalin fixed, paraffin wax embedded sections from 20 cases of Hodgkin's disease (six mixed cellularity, 14 nodular sclerosis) were immunostained for CD15, using two antibodies (DAKO-M1 and Leu-M1) and for CD30 using the antibody Ber-H2. The staining was carried out by conventional techniques which included pretreatment of sections with trypsin and on untreated sections following heating with microwaves. With antibody Leu-M1 an additional method, using a specific antimouse IgM bridge both with and without microwave heating, was also included. The results for each method were compared by counting positively stained Reed-Sternberg cells and estimating the staining intensity.
Results:
Microwave heating resulted in a substantial increase in the number of cells stained with antibodies to CD15 and also in the staining intensity. The best results were obtained using Leu-M1 with specific rabbit anti-mouse IgM bridge and microwave heating. Dramatic enhancement of the staining of Reed-Sternberg cells for CD30 was achieved following microwave heating, together with disappearance of the non-specific staining of plasma cells.
Conclusion:
Microwave heating is strongly recommended for the immunohistochemical staining of CD15 and CD30 expressed by Reed-Sternberg cells in Hodgkin's disease.
