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.
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.
Authors: H Stein; D Y Mason; J Gerdes; N O'Connor; J Wainscoat; G Pallesen; K Gatter; B Falini; G Delsol; H Lemke Journal: Blood Date: 1985-10 Impact factor: 22.113
Authors: R von Wasielewski; M Mengel; R Fischer; M L Hansmann; K Hübner; J Franklin; H Tesch; U Paulus; M Werner; V Diehl; A Georgii Journal: Am J Pathol Date: 1997-10 Impact factor: 4.307