Rapid Frozen Tissue Immunostaining of Melanocytes in Chronically Photodamaged Skin - PowerPoint PPT Presentation

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Rapid Frozen Tissue Immunostaining of Melanocytes in Chronically Photodamaged Skin

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Title: Rapid Frozen Tissue Immunostaining of Melanocytes in Chronically Photodamaged Skin


1
Rapid Frozen Tissue Immunostaining of Melanocytes
in Chronically Photodamaged Skin
  • Basil Cherpelis MD, Jane L Messina, C. Wayne
    Cruse, Graham S. Clark, MD , Neil A Fenske, Ren
    Chen, M.D and Frank Glass MD

University of South Florida Dermatology and
Cutaneous Surgery, Tampa, FL. Division of
Cutaneous Oncology H Lee Moffitt Cancer Center
and Research Institute
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Excision of LM
3
Surgical margins for LM
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Agarwal-Antal, N., G.M. Bowen, and J.W. Gerwels,
J Am Acad Dermatol, 2002. 47(5) p. 743-8
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  • 2 mm vertical strips 5 mm outlined
  • using geometric configuration
  • 2. 2 mm strip 2 mm around positive margins

J Surg Oncol, 2005. 91(2) p. 120-5
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Surgical Management of Melanoma in Situ Using A
Staged Marginal and Central Excision
Technique Mecker G. Möller, MD, Effie
Pappas-Politis, MD, Jonathan S. Zager, MD, Luis
A. Santiago, MD, Daohai Yu, PhD, Amy Prakash, MD,
Adam Kinal, BS, Graham S. Clark, BS, Weiwei Zhu,
MS, Christopher A. Puleo, PA-C, L. Frank Glass,
MD, Jane L Messina, MD, Vernon K. Sondak, MD, C.
Wayne Cruse, MD
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N49
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Recurrence Rates for LMMohs Micrographic Surgery
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  • Frozen section sensitivity on HE 100,
    specificity 90
  • IHC for confirmation or refutation
  • 92.5 clear margins one layer
  • Distinction of keratinocytes from melanocytes
    simplified
  • but melanocytic hyperplasia sun damaged skin
    difficult

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  • 11 ½ minute protocol
  • Conjugating 2nd antibody to a dextran polymer
  • backbone and horseradish peroxidase
  • Azure blue and DAB

17
Melanocyte
Polymer
Antigen-Antibody Link
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25 patients, 100 slides
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MART-1 paraffin vs. frozen Simple t-test was used
to test whether paraffin and frozen will give
significant different results in the
measurements.
  • Conclusion Simple t-test, which compares every
    measurement obtained from the two different
    methods, no significant difference was found.

22
MART frozen vs paraffin - MANOVA results
Conclusion There is no significant difference in
any of the five measurements that were obtained
from the two methods MART-1 frozen or paraffin.
23
Long standing sun exposed skin
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All of the 4 methods were compared were based on
general linear regression.
Conclusion Except Melanocyte Nucleus diameter
(microns), the results from the 4 methods have no
significant difference. (f value 0.06, p value
0.807)
26
HE Frozen vs Paraffin simple t-test
Conclusion by simple t-test, which compares
every measurement that got from the two different
methods, no significant difference was found
27
HE Frozen vs Paraffin - MANOVA results
Conclusion there is no significant difference in
any of the five measurements that were got from
the two methods.
28
  • Frozen sections of sun damaged skin equivalent
    with MART-1 permanent sections

MART-1 also stains normal dendritic melanocytes
in the background. Contiguous melanocytes Nesting
Melanocyte atypia Follicular involvement
29
Frozen tissue immunostaining of melanocytes
microphthalmia-associated transcription factor
in chronically photodamaged skinLF Glass,
Higgins II MA, GS Clark MD, B Cherpelis MD, S
Ladd BS, HTL (ASCP), NA Fenske
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a
Frozen MITF
b
Permanent
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.
a no significant difference (p0.5101) b no
significnat difference by Wilcoxon signed rank
test (p0.0735).
32
Melanocyte Diameter and Density in
Melanoma in Situ and Solar Lentigines
Involving Chronically Sundamaged Skin by MITF
HW Higgins II MA, GS Clark MD, B Cherpelis MD,
S Ladd BS, HTL (ASCP), LF Glass MD
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MITF
Solar Lentigo versus MMIS
independent t-test
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MITF
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Density
MITF
Tukey multiple comparison tests
Diameter
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D X D
MITF
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MITF
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Density (/200 microns)
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Diameter (microns)
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Diameter X Density
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Summary
  • Melanocyte parameters measured by MITF in
    solar lentigines significantly different
    than MIS, but equivalent to melanocytic
    hyperplasia of chronically
    sundamaged skin
  • MITF is a nuclear stain, a tool to locate and
    quantify melanocytes in both permanent and frozen
    sections with similar results
  • MITF produces similar results as MART-1 for
    nuclear diameter, but melanocyte density is less

43
19 Minute Cytokeratin Immunostaining of Basal and
Squamous Cell Carcinoma in Mohs Micrographic
Surgery
  • Logan Turner MD, Basil Cherpelis MD, Jane L
    Messina, C. Wayne Cruse, Graham S. Clark, MD ,
    Neil A Fenske, and Frank Glass MD

University of South Florida Dermatology and
Cutaneous Surgery, Tampa, FL. Division of
Cutaneous Oncology H Lee Moffitt Cancer Center
and Research Institute
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Atypical Keratinocyte
CK
Polymer
Antigen-Antibody Link
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Frozen HE
Perm HE
CK
CK
Ber-EP4
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Case 1
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AE1/AE3
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AE1/AE3
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Case 2
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  • AE1/AE3 confirmed free margins, avoiding
    sacrifice of tissue
  • 36/400 (9), 18 dense infiltrates (found tumor
    in 3 cases)
  • 18 perineural involvement, morpheaform BCCs
    (helpful in 8 cases)

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Rapid (19 Minute) Cytokeratin Immunostaining of
Basal and Squamous Cell Carcinoma in Mohs
Micrographic Surgery
Summary
  • Rapid 19 min protocol for AE1/AE3 gives quality
    results
  • Frozen technique uses traditional methods of
    IHC
  • Cost low (30.00/slide, 88342 reimburses
    45.31)
  • Margins assured when dense infiltrates are
    present, and if scarring is extensive.
  • Also aggressive growth tumors
  • Perineural spread
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