Title: Rapid Frozen Tissue Immunostaining of Melanocytes in Chronically Photodamaged Skin
1Rapid 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
2Excision of LM
3Surgical margins for LM
4Agarwal-Antal, N., G.M. Bowen, and J.W. Gerwels,
J Am Acad Dermatol, 2002. 47(5) p. 743-8
5- 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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9Surgical 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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12N49
13Recurrence Rates for LMMohs Micrographic Surgery
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15- 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
16- 11 ½ minute protocol
- Conjugating 2nd antibody to a dextran polymer
- backbone and horseradish peroxidase
- Azure blue and DAB
17Melanocyte
Polymer
Antigen-Antibody Link
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2025 patients, 100 slides
21MART-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.
22MART 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.
23Long standing sun exposed skin
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25All 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)
26HE 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
29Frozen 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
30a
Frozen MITF
b
Permanent
31.
a no significant difference (p0.5101) b no
significnat difference by Wilcoxon signed rank
test (p0.0735).
32Melanocyte 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
33MITF
Solar Lentigo versus MMIS
independent t-test
34MITF
35Density
MITF
Tukey multiple comparison tests
Diameter
36D X D
MITF
37MITF
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39Density (/200 microns)
40Diameter (microns)
41Diameter X Density
42Summary
- 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
4319 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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47Atypical Keratinocyte
CK
Polymer
Antigen-Antibody Link
48Frozen HE
Perm HE
CK
CK
Ber-EP4
49Case 1
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51AE1/AE3
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53AE1/AE3
54Case 2
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59- 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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61Rapid (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