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Evaluation of digital staining for ex vivo confocal laser scanning microscopy.

Abstract

BACKGROUND

Ex vivo confocal laser scanning microscopy (CLSM) is a novel diagnostic tool for the fast examination of native tissue. However, CLSM produces black/white/green images, depending on the refraction indices of the tissue structures, complemented by nuclear fluorescence staining, which the vast majority of Mohs surgeons and dermatopathologists are not trained to interpret. Digital staining is applicable to ex vivo CLSM investigations to simulate the images of conventional slides stained with haematoxylin & eosin (H&E).

OBJECTIVES

The aim of our study was to evaluate in detail the appearance of human skin structures using digitally stained ex vivo CLSM images and compare the results to that of conventional H&E slides of the same specimen.

METHODS

After providing informed consent, 26 patients donated their Burow's triangles (healthy skin) that resulted from plastic reconstruction after the R0- excision of skin tumours. After being investigated by ex vivo CLSM, including automated digital staining (VivaScope 2500M-4G, MAVIG GmbH), the specimens were fixed in formalin, embedded in paraffin and stained with H&E.

RESULTS

Almost all skin structures in the digitally stained ex vivo CLSM images morphologically resembled the structures in the histopathological images acquired from H&E slides. Due to the high refraction index of melanin, the hair shafts appeared bright pink, and the melanocytes and melanophages were poorly imaged, resulting in a strong pink appearance that vastly differed from the appearance of conventional H&E-stained histopathology.

CONCLUSIONS

Digital staining of ex vivo CLSM images is an easy and highly useful tool to facilitate the interpretation of black-field images generated by confocal laser scanning microscopy for dermatopathologists and Mohs surgeons who are familiar with H&E staining. Unlike the pigmented structures, the cutaneous and subcutaneous structures had excellent visualization with only minimal differences from their appearance on H&E slides.

Authors+Show Affiliations

University of Leipzig, Department of Dermatology, Venerology and Allergology.University of Leipzig, Department of Dermatology, Venerology and Allergology.University of Leipzig, Department of Dermatology, Venerology and Allergology.University of Leipzig, Department of Dermatology, Venerology and Allergology.University of Leipzig, Department of Dermatology, Venerology and Allergology.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31732988

Citation

Schüürmann, M, et al. "Evaluation of Digital Staining for Ex Vivo Confocal Laser Scanning Microscopy." Journal of the European Academy of Dermatology and Venereology : JEADV, 2019.
Schüürmann M, Stecher MM, Paasch U, et al. Evaluation of digital staining for ex vivo confocal laser scanning microscopy. J Eur Acad Dermatol Venereol. 2019.
Schüürmann, M., Stecher, M. M., Paasch, U., Simon, J. C., & Grunewald, S. (2019). Evaluation of digital staining for ex vivo confocal laser scanning microscopy. Journal of the European Academy of Dermatology and Venereology : JEADV, doi:10.1111/jdv.16085.
Schüürmann M, et al. Evaluation of Digital Staining for Ex Vivo Confocal Laser Scanning Microscopy. J Eur Acad Dermatol Venereol. 2019 Nov 15; PubMed PMID: 31732988.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of digital staining for ex vivo confocal laser scanning microscopy. AU - Schüürmann,M, AU - Stecher,M M, AU - Paasch,U, AU - Simon,J C, AU - Grunewald,S, Y1 - 2019/11/15/ PY - 2019/11/17/entrez KW - dermatopathology KW - digital staining KW - ex vivo confocal laser scan microscopy JF - Journal of the European Academy of Dermatology and Venereology : JEADV JO - J Eur Acad Dermatol Venereol N2 - BACKGROUND: Ex vivo confocal laser scanning microscopy (CLSM) is a novel diagnostic tool for the fast examination of native tissue. However, CLSM produces black/white/green images, depending on the refraction indices of the tissue structures, complemented by nuclear fluorescence staining, which the vast majority of Mohs surgeons and dermatopathologists are not trained to interpret. Digital staining is applicable to ex vivo CLSM investigations to simulate the images of conventional slides stained with haematoxylin & eosin (H&E). OBJECTIVES: The aim of our study was to evaluate in detail the appearance of human skin structures using digitally stained ex vivo CLSM images and compare the results to that of conventional H&E slides of the same specimen. METHODS: After providing informed consent, 26 patients donated their Burow's triangles (healthy skin) that resulted from plastic reconstruction after the R0- excision of skin tumours. After being investigated by ex vivo CLSM, including automated digital staining (VivaScope 2500M-4G, MAVIG GmbH), the specimens were fixed in formalin, embedded in paraffin and stained with H&E. RESULTS: Almost all skin structures in the digitally stained ex vivo CLSM images morphologically resembled the structures in the histopathological images acquired from H&E slides. Due to the high refraction index of melanin, the hair shafts appeared bright pink, and the melanocytes and melanophages were poorly imaged, resulting in a strong pink appearance that vastly differed from the appearance of conventional H&E-stained histopathology. CONCLUSIONS: Digital staining of ex vivo CLSM images is an easy and highly useful tool to facilitate the interpretation of black-field images generated by confocal laser scanning microscopy for dermatopathologists and Mohs surgeons who are familiar with H&E staining. Unlike the pigmented structures, the cutaneous and subcutaneous structures had excellent visualization with only minimal differences from their appearance on H&E slides. SN - 1468-3083 UR - https://www.unboundmedicine.com/medline/citation/31732988/Evaluation_of_digital_staining_for_ex_vivo_confocal_laser_scanning_microscopy L2 - https://doi.org/10.1111/jdv.16085 DB - PRIME DP - Unbound Medicine ER -