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Rapid Frozen Section Immunostaining of Melanocytes by Microphthalmia-Associated Transcription Factor

Glass, L Frank MD; Raziano, Renata M MD, PhD; Clark, Graham S MD; Higgins, H William MA; Ladd, Sharron BS, HTL, (ASCP); Lien, Mary H MD; Chen, Ren MD, MPH; Cherpelis, Basil S MD

The American Journal of Dermatopathology: June 2010 - Volume 32 - Issue 4 - p 319-325
doi: 10.1097/DAD.0b013e3181bcd94f
Original Study

Mohs micrographic surgery (MMS) has increasingly become an accepted therapy for melanoma in situ on chronically sun damaged skin (CSDS). However, melanocytes are difficult to locate in frozen material on hematoxylin and eosin. In addition, determining the cut-off between the melanoma and the “atypical melanocytic hyperplasia” in CSDS can be challenging in frozen or formalin-fixed paraffin-embedded sections, with or without immunohistochemistry (IHC). In this article, we report the use of a rapid, 35-minute protocol using microphthalmia-associated transcription factor (MITF) IHC for identifying melanocytes in frozen tissue for its potential use in MMS. In contrast to melanoma antigen recognized by T cells (MART-1), MITF is a nuclear stain, which simplifies identification of melanocytes and quantification of melanocytic parameters. In this study, MITF IHC in frozen sections yielded equivalent melanocyte nuclear diameter and density measurements compared with formalin-fixed paraffin-embedded sections. Nuclear diameter measurements obtained with MITF were similar to that previously reported with MART-1, but the melanocyte density figures were lower. Reliable labeling of melanocytes in frozen sections required the use of diaminobenzidine (DAB) chromogen with Giemsa counterstaining and a buffer devoid of surfactant. Our experience with MITF IHC indicates that it is a dependable immunostain in frozen sections, and may prove to be useful in MMS as an adjunct to hematoxylin and eosin and MART-1 IHC for interpretation of margins for melanoma in situ on CSDS.

From the *Department of Dermatology and Cutaneous Surgery, University of South Florida, Tampa, FL; †Department of Dermatology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL; ‡Department of Dermatology, James A. Haley Veterans' Hospital, Tampa, FL; §Florida State University College of Medicine, Tallahassee, FL; and ¶Office of Research, Biostatistics Core, University of South Florida, Tampa, FL.

Funding sources: none.

Presented in part at the following poster sessions: 45th Annual Meeting of the American Society of Dermatopathology, October 16-18, 2008, San Francisco, CA, and at 2nd World Meeting of Interdisciplinary Melanoma/Skin Cancer Centers, November, 20-23, 2008, Clearwater Beach, FL.

Disclosures: Dr. Lien is a speaker for Abbott Laboratories, Abbott Park, IL.

Published in abstract form in: Calder K, Higgins H, Clark GS, Cherpelis BS, Ladd S, Glass LF. Rapid frozen tissue immunostaining of melanocytes by microophthalmic transcription factor. J Cutan Pathol. 2009;36:180.

Reprints: L. Frank Glass, MD, Department of Dermatology and Cutaneous Surgery, University of South Florida, 12901 Bruce B. Downs, MDC 79 (MDA 1174), Tampa, FL (e-mail:

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