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Ex-vivo konfokale Laserscanmikroskopie bei melanozytären Läsionen und Autoimmunerkrankungen

Ex vivo confocal laser scanning microscopy for melanocytic lesions and autoimmune diseases

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Zusammenfassung

Hintergrund

Die Ex-vivo-KLM (konfokale Laserscanmikroskopie) ermöglicht die Histologie am Krankenbett und bietet dem Chirurgen eine direkte intraoperative Gewebeuntersuchung.

Ziel der Arbeit

Ziel war es herauszufinden, ob dieses innovative, ultraschnelle Diagnoseinstrument neben nichtmelanozytären Hautkrebsformen, insbesondere dem Basalzellkarzinom, auch bei anderen Indikationen, wie melanozytären Läsionen und Autoimmunerkrankungen, nutzbringend eingesetzt werden kann.

Material und Methoden

Literaturbeurteilung und Zusammenfassung der derzeitigen Kenntnisse und Erfahrungen zum Einsatz der Ex-vivo-KLM bei melanozytären Läsionen und bei Autoimmunerkrankungen.

Ergebnisse

Die bisherigen Erfahrungen mit der Anwendung der Ex-vivo-KLM bei melanozytären Läsionen und bei Autoimmunerkrankungen sind begrenzt, zeigen aber vielversprechende Perspektiven. Es werden derzeitige Kenntnisse und Erfahrungen zum Einsatz der Ex-vivo-KLM bei melanozytären Hautveränderungen sowie Beispiele mit den klassischen Ex-vivo-KLM-Merkmalen vorgestellt. Präsentiert werden ferner die bisherigen Ergebnisse zum Einsatz der Ex-vivo-KLM bei entzündlichen Autoimmundermatosen.

Diskussion

Die Methode eignet sich insbesondere zur Schnellschnittuntersuchung von Basalzellkarzinomen während der Mohs-Chirurgie, könnte aber in Zukunft auch bei der intraoperativen Untersuchung von melanozytären Hautveränderungen und Autoimmunerkrankungen eingesetzt werden.

Abstract

Background

Ex vivo confocal laser scanning microscopy (CLSM) enables bedside histology and offers the surgeon a direct intraoperative tissue examination.

Objectives

To determine whether this innovative, ultra-fast diagnostic tool can be expanded beyond nonmelanoma skin cancer, particularly basal cell carcinoma, to other indications including melanocytic lesions and autoimmune diseases.

Materials and methods

Review of literature and summary of the current knowledge and experience of the use of ex vivo CLSM in melanocytic lesions and in autoimmune diseases.

Results

Up to date experience of the use of ex vivo CLSM in melanocytic lesions and in autoimmune diseases is limited but promising. Current knowledge on melanocytic lesions in ex vivo CLSM and their examples together with classic ex vivo CLSM features are presented. Previous results on the use of ex vivo CLSM in autoimmune dermatoses are presented, and future application possibilities of ex vivo CLSM are discussed.

Conclusions

The method is particularly suitable for the rapid examination of basal cell carcinomas during Mohs surgery but could also be used in the future for the intraoperative examination of melanocytic and autoimmune skin lesions.

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Literatur

  1. Hartmann D, Ruini C, Mathemeier L, Dietrich A, Ruzicka T, von Braunmühl T (2016) Identification of ex-vivo confocal scanning microscopic features and their histological correlates in human skin. J Biophotonics 9:376–387

    PubMed  Google Scholar 

  2. Maier T, Sattler EC, Braun-Falco M, Korting HC, Ruzicka T, Berking C (2013) Reflectance confocal microscopy in the diagnosis of partially and completely amelanotic melanoma: report on seven cases. J Eur Acad Dermatol Venereol 27:e42–e52

    CAS  PubMed  Google Scholar 

  3. Sattler EC, Maier T, Hoffmann VS, Hegyi J, Ruzicka T, Berking C (2012) Noninvasive in vivo detection and quantification of Demodex mites by confocal laser scanning microscopy. Br J Dermatol 167:1042–1047

    CAS  PubMed  Google Scholar 

  4. Longo C, Casari A, Beretti F, Cesinaro AM, Pellacani G (2013) Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy. J Am Acad Dermatol 68:e73–e82

    PubMed  Google Scholar 

  5. Ulrich M, Maltusch A, Röwert-Huber J, González S, Sterry W, Stockfleth E, Astner S (2007) Actinic keratoses: non-invasive diagnosis for field cancerisation. Br J Dermatol 156(Suppl 3):13–17

    PubMed  Google Scholar 

  6. Pellacani G, Scope A, Ferrari B, Pupelli G, Bassoli S, Longo C, Cesinaro AM, Argenziano G, Hofmann-Wellenhof R, Malvehy J, Marghoob AA, Puig S, Seidenari S, Soyer HP, Zalaudek I (2009) New insights into nevogenesis: in vivo characterization and follow-up of melanocytic nevi by reflectance confocal microscopy. J Am Acad Dermatol 61:1001–1013

    PubMed  Google Scholar 

  7. Ardigo M, Cota C, Berardesca E, González S (2009) Concordance between in vivo reflectance confocal microscopy and histology in the evaluation of plaque psoriasis. J Eur Acad Dermatol Venereol 23:660–667

    CAS  PubMed  Google Scholar 

  8. Longo C, Casari A, De Pace B, Simonazzi S, Mazzaglia G, Pellacani G (2013) Proposal for an in vivo histopathologic scoring system for skin aging by means of confocal microscopy. Skin Res Technol 19:e167–e173

    PubMed  Google Scholar 

  9. Ulrich M, Lange-Asschenfeldt S, Gonzalez S (2012) The use of reflectance confocal microscopy for monitoring response to therapy of skin malignancies. Dermatol Pract Concept 2:202a10

    PubMed  Google Scholar 

  10. Manfredini M, Mazzaglia G, Ciardo S, Farnetani F, Mandel VD, Longo C, Zauli S, Bettoli V, Virgili A, Pellacani G (2015) Acne: in vivo morphologic study of lesions and surrounding skin by means of reflectance confocal microscopy. J Eur Acad Dermatol Venereol 29:933–939

    CAS  PubMed  Google Scholar 

  11. Rajadhyaksha M, Grossman M, Esterowitz D, Webb RH, Anderson RR (1995) In vivo confocal scanning laser microscopy of human skin: melanin provides strong contrast. J Invest Dermatol 104:946–952

    CAS  PubMed  Google Scholar 

  12. Rajadhyaksha M, González S, Zavislan JM, Anderson RR, Webb RH (1999) In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology. J Invest Dermatol 113:293–303

    CAS  PubMed  Google Scholar 

  13. Pellacani G, Farnetani F, Gonzalez S, Longo C, Cesinaro AM, Casari A, Beretti F, Seidenari S, Gill M (2012) In vivo confocal microscopy for detection and grading of dysplastic nevi: a pilot study. J Am Acad Dermatol 66:e109–e121

    PubMed  Google Scholar 

  14. Ulrich M, Lange-Asschenfeldt S, Gonzalez S (2012) Clinical applicability of in vivo reflectance confocal microscopy in dermatology. G Ital Dermatol Venereol 147:171–178

    CAS  PubMed  Google Scholar 

  15. Longo C, Farnetani F, Ciardo S, Cesinaro AM, Moscarella E, Ponti G, Zalaudek I, Argenziano G, Pellacani G (2013) Is confocal microscopy a valuable tool in diagnosing nodular lesions? A study of 140 cases. Br J Dermatol 169:58–67

    CAS  PubMed  Google Scholar 

  16. Pellacani G, Pepe P, Casari A, Longo C (2014) Reflectance confocal microscopy as a second-level examination in skin oncology improves diagnostic accuracy and saves unnecessary excisions: a longitudinal prospective study. Br J Dermatol 171:1044–1051

    CAS  PubMed  Google Scholar 

  17. Hoogedoorn L, Peppelman M, van de Kerkhof PC, van Erp PE, Gerritsen MJ (2015) The value of in vivo reflectance confocal microscopy in the diagnosis and monitoring of inflammatory and infectious skin diseases: a systematic review. Br J Dermatol 172:1222–1248

    CAS  PubMed  Google Scholar 

  18. Longo C, Lallas A, Kyrgidis A, Rabinovitz H, Moscarella E, Ciardo S, Zalaudek I, Oliviero M, Losi A, Gonzalez S, Guitera P, Piana S, Argenziano G, Pellacani G (2014) Classifying distinct basal cell carcinoma subtype by means of dermatoscopy and reflectance confocal microscopy. J Am Acad Dermatol 71:716–724.e1

    PubMed  Google Scholar 

  19. Segura S, Puig S, Carrera C, Palou J, Malvehy J (2009) Development of a two-step method for the diagnosis of melanoma by reflectance confocal microscopy. J Am Acad Dermatol 61:216–229

    PubMed  Google Scholar 

  20. Guitera P, Menzies SW, Longo C, Cesinaro AM, Scolyer RA, Pellacani G (2012) In vivo confocal microscopy for diagnosis of melanoma and basal cell carcinoma using a two-step method: analysis of 710 consecutive clinically equivocal cases. J Invest Dermatol 132:2386–2394

    CAS  PubMed  Google Scholar 

  21. Gareau DS (2009) Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology. J Biomed Opt 14:34050

    PubMed  Google Scholar 

  22. Jonak C, Skvara H, Kunstfeld R, Trautinger F, Schmid JA (2011) Intradermal indocyanine green for in vivo fluorescence laser scanning microscopy of human skin: a pilot study. Plos One 6:e23972

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Goetz M, Toermer T, Vieth M, Dunbar K, Hoffman A, Galle PR, Neurath MF, Delaney P, Kiesslich R (2009) Simultaneous confocal laser endomicroscopy and chromoendoscopy with topical cresyl violet. Gastrointest Endosc 70:959–968

    PubMed  Google Scholar 

  24. Al-Arashi MY, Salomatina E, Yaroslavsky AN (2007) Multimodal confocal microscopy for diagnosing nonmelanoma skin cancers. Lasers Surg Med 39:696–705

    PubMed  Google Scholar 

  25. Yaroslavsky AN, Barbosa J, Neel V, DiMarzio C, Anderson RR (2005) Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer. J Biomed Opt 10:14011

    PubMed  Google Scholar 

  26. Kaeb S, Landthaler M, Hohenleutner U (2009) Confocal laser scanning microscopy—evaluation of native tissue sections in micrographic surgery. Lasers Med Sci 24:819–823

    PubMed  Google Scholar 

  27. Longo C, Rajadhyaksha M, Ragazzi M, Nehal K, Gardini S, Moscarella E, Lallas A, Zalaudek I, Piana S, Argenziano G, Pellacani G (2014) Evaluating ex vivo fluorescence confocal microscopy images of basal cell carcinomas in Mohs excised tissue. Br J Dermatol 171:561–570

    CAS  PubMed  Google Scholar 

  28. Longo C, Ragazzi M, Castagnetti F, Gardini S, Palmieri T, Lallas A, Moscarella E, Piana S, Pellacani G, Zalaudek I, Argenziano G (2013) Inserting ex vivo fluorescence confocal microscopy perioperatively in Mohs micrographic surgery expedites bedside assessment of excision margins in recurrent basal cell carcinoma. Dermatology 227:89–92

    PubMed  Google Scholar 

  29. Bennàssar A, Carrera C, Puig S, Vilalta A, Malvehy J (2013) Fast evaluation of 69 basal cell carcinomas with ex vivo fluorescence confocal microscopy: criteria description, histopathological correlation, and interobserver agreement. JAMA Dermatol 149:839–847

    PubMed  Google Scholar 

  30. Gareau DS, Karen JK, Dusza SW, Tudisco M, Nehal KS, Rajadhyaksha M (2009) Sensitivity and specificity for detecting basal cell carcinomas in Mohs excisions with confocal fluorescence mosaicing microscopy. J Biomed Opt 14:34012

    PubMed  Google Scholar 

  31. Karen JK, Gareau DS, Dusza SW, Tudisco M, Rajadhyaksha M, Nehal KS (2009) Detection of basal cell carcinomas in Mohs excisions with fluorescence confocal mosaicing microscopy. Br J Dermatol 160:1242–1250

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Patel YG, Nehal KS, Aranda I, Li Y, Halpern AC, Rajadhyaksha M (2007) Confocal reflectance mosaicing of basal cell carcinomas in Mohs surgical skin excisions. J Biomed Opt 12:34027

    PubMed  Google Scholar 

  33. Perez-Anker J, Puig S, Malvehy J (2020) A fast and effective option for tissue flattening: optimizing time and efficacy in ex vivo confocal microscopy. J Am Acad Dermatol 82:e157–e158

    PubMed  Google Scholar 

  34. Pellacani G, Cesinaro AM, Seidenari S (2005) Reflectance-mode confocal microscopy for the in vivo characterization of pagetoid melanocytosis in melanomas and nevi. J Invest Dermatol 125:532–537

    CAS  PubMed  Google Scholar 

  35. Scope A, Benvenuto-Andrade C, Agero AL, Halpern AC, Gonzalez S, Marghoob AA (2007) Correlation of dermoscopic structures of melanocytic lesions to reflectance confocal microscopy. Arch Dermatol 143:176–185

    PubMed  Google Scholar 

  36. Pellacani G, Longo C, Malvehy J, Puig S, Carrera C, Segura S, Bassoli S, Seidenari S (2008) In vivo confocal microscopic and histopathologic correlations of dermoscopic features in 202 melanocytic lesions. Arch Dermatol 144:1597–1608

    PubMed  Google Scholar 

  37. Scope A, Benvenuto-Andrade C, Agero AL, Malvehy J, Puig S, Rajadhyaksha M, Busam KJ, Marra DE, Torres A, Propperova I, Langley RG, Marghoob AA, Pellacani G, Seidenari S, Halpern AC, Gonzalez S (2007) In vivo reflectance confocal microscopy imaging of melanocytic skin lesions: consensus terminology glossary and illustrative images. J Am Acad Dermatol 57:644–658

    PubMed  Google Scholar 

  38. Gill M, Longo C, Farnetani F, Cesinaro AM, González S, Pellacani G (2014) Non-invasive in vivo dermatopathology: identification of reflectance confocal microscopic correlates to specific histological features seen in melanocytic neoplasms. J Eur Acad Dermatol Venereol 28:1069–1078

    CAS  PubMed  Google Scholar 

  39. Pellacani G, Cesinaro AM, Seidenari S (2005) Reflectance-mode confocal microscopy of pigmented skin lesions—improvement in melanoma diagnostic specificity. J Am Acad Dermatol 53:979–985

    PubMed  Google Scholar 

  40. Gerger A, Koller S, Weger W, Richtig E, Kerl H, Samonigg H, Krippl P, Smolle J (2006) Sensitivity and specificity of confocal laser-scanning microscopy for in vivo diagnosis of malignant skin tumors. Cancer 107:193–200

    PubMed  Google Scholar 

  41. Pellacani G, Guitera P, Longo C, Avramidis M, Seidenari S, Menzies S (2007) The impact of in vivo reflectance confocal microscopy for the diagnostic accuracy of melanoma and equivocal melanocytic lesions. J Invest Dermatol 127:2759–2765

    CAS  PubMed  Google Scholar 

  42. Bennàssar A, Vilata A, Puig S, Malvehy J (2014) Ex vivo fluorescence confocal microscopy for fast evaluation of tumour margins during Mohs surgery. Br J Dermatol 170:360–365

    PubMed  Google Scholar 

  43. Hartmann D, Ruini C, Mathemeier L, Bachmann MR, Dietrich A, Ruzicka T, von Braunmühl T (2017) Identification of ex-vivo confocal laser scanning microscopic features of melanocytic lesions and their histological correlates. J Biophotonics 10:128–142

    PubMed  Google Scholar 

  44. Hartmann D, Krammer S, Ruini C, Ruzicka T, von Braunmühl T (2016) Correlation of histological and ex-vivo confocal tumor thickness in malignant melanoma. Lasers Med Sci 31:921–927

    PubMed  Google Scholar 

  45. Hartmann D, Krammer S, Vural S, Bachmann MR, Ruini C, Sárdy M, Ruzicka T, Berking C, von Braunmühl T (2018) Immunofluorescence and confocal microscopy for ex-vivo diagnosis of melanocytic and non-melanocytic skin tumors: a pilot study. J Biophotonics 11. https://doi.org/10.1002/jbio.201700211

    Article  PubMed  Google Scholar 

  46. Bağcı IS, Aoki R, Krammer S, Ruzicka T, Sárdy M, Hartmann D (2019) Ex vivo confocal laser scanning microscopy: An innovative method for direct immunofluorescence of cutaneous vasculitis. J Biophotonics 12:e201800425

    PubMed  Google Scholar 

  47. Bağcı IS, Aoki R, Krammer S, Ruzicka T, Sárdy M, French LE, Hartmann D (2019) Ex vivo confocal laser scanning microscopy for bullous pemphigoid diagnostics: new era in direct immunofluorescence? J Eur Acad Dermatol Venereol 33:2123–2130

    PubMed  Google Scholar 

  48. Bağcı IS, Aoki R, Krammer S, Vladimirova G, Ruzicka T, Sárdy M, French LE, Hartmann D (2020) Immunofluorescence and histopathological assessment using ex vivo confocal laser scanning microscopy in lichen planus. J Biophotonics 13:e202000328

    PubMed  Google Scholar 

  49. Krammer S, Krammer C, Salzer S, Bağci IS, French LE, Hartmann D (2019) Recurrence of pemphigus vulgaris under nivolumab therapy. Front Med 6:262

    Google Scholar 

  50. Bağcı IS, Aoki R, Vladimirova G, Sárdy M, Ruzicka T, French LE, Hartmann D (2021) Simultaneous immunofluorescence and histology in pemphigus vulgaris using ex vivo confocal laser scanning microscopy. J Biophotonics 14:e202000509

    PubMed  Google Scholar 

  51. Bağcı SI, Aoki R, Vladimirova G, Ergün E, Ruzicka T, Sárdy M, French LE, Hartmann D (2021) New-generation diagnostics in inflammatory skin diseases: Immunofluorescence and histopathological assessment using ex vivo confocal laser scanning microscopy in cutaneous lupus erythematosus. Exp Dermatol 30:684–690

    Google Scholar 

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Hartmann, D. Ex-vivo konfokale Laserscanmikroskopie bei melanozytären Läsionen und Autoimmunerkrankungen. Hautarzt 72, 1058–1065 (2021). https://doi.org/10.1007/s00105-021-04906-1

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