The Potential Use of Elastic Tissue Autofluorescence in Formalin- fixed Paraffin-embedded Skin Biopsies

Autofluorescence (AF) or naïve-florescence is the natural emission of light by biomolecules. During florescence microscope examination, we realized that elastic tissue is brighter or more autoflourescent than collagen and other biomolecules/cells in the skin. Consequently, we decided to review elastic tissue-related pathologies under a florescence microscope and to report the possible benefits of this technique from selected cases from the paraffin-block archive, by using the protease digestion immunofluorescence method. Selected and clinic- pathologically confirmed 3 elastofibroma dorsi, 3 pseudoxanthoma elasticum, 3 anetoderma, 3 arteriovenous malformations, 3 temporal arteritis, 3 scar tissue and 3 highly solar-damaged samples of skin from 2014-2019 were retrieved. Under the fluorescent microscope, coarse, thick and globularly-fragmented elastic fibers of elastofibroma dorsi, shortened, irregular and convoluted elastic fibers of pseudoxanthoma elasticum, internal elastic membranes of arteries and their integrity was visualized. None of the anetoderma cases had any signal representing elastic tissue. It was shown that elastic tissue can be observed easily under fluorescence microscope in the case of FFPE tissues. The resulting autofluorescence can be useful in recognizing elastic tissue-related pathologies, and it may be used as an ancillary or an alternative method to routine histochemical techniques.

PDF

___

1] Nasr SH, Galgano SJ, Markowitz GS et al. Immunofluorescence on pronase-digested paraffin sections: a valuable salvage technique for renal biopsies. Kidney Int. 2006;70(12):2148-51. https://doi.org/10.1038/ sj.ki.5001990
[2] Nasr SH, Fidler ME, Said SM. Paraffin Immunofluorescence: A Valuable Ancillary Technique in Renal Pathology. Kidney Int Rep. 2018;3(6):1260-6. https://doi.org/10.1016/j. ekir.2018.07.008
[3] Valencia-Guerrero A, Deng A, Dresser K et al. The Value of Direct Immunofluorescence on Proteinase-Digested Formalin-Fixed Paraffin-Embedded Skin Biopsies. Am J Dermatopathol. 2018;40(2):111-7. https://doi. org/10.1097/DAD.0000000000000934
[4] Borucki R, Perry DM, Lopez-Garcia DR et al. Fluorescence microscopy for the evaluation of elastic tissue patterns within fibrous proliferations of the skin on hematoxylin-eosin-stained slides. J Am Acad Dermatol. 2018;S0190-9622(18)30016-1. https://doi.org/10.1016/j. jaad.2017.12.073
[5] Elston DM. Elastic Fibers in Scars and Alopecia. Am J Dermatopathol. 2017;39(7):556-557. https://doi. org/10.1097/DAD.0000000000000712
[6] Basko-Plluska J, Kazlouskaya V, Spizuoco A et al. The use of fluorescence microscopy to evaluate elastic fiber pattern in melanocytic neoplasms. Am J Dermatopathol. 2014;36(5):443-4. https://doi.org/10.1097/ DAD.0b013e31829b219b
[7] Elston CA, Kazlouskaya V, Elston DM. Elastic staining versus fluorescent and polarized microscopy in the diagnosis of alopecia. J Am Acad Dermatol. 2013;69(2):288-93. doi: https://doi.org/10.1016/j.jaad.2013.02.030. Epub 2013 May 14.
[8] Elston DM. Medical Pearl: fluorescence microscopy of hematoxylin-eosin-stained sections. J Am Acad Dermatol. 2002;47(5):777-9. https://doi.org/10.1067/ mjd.2002.120623
[9] Xie H, Su H, Chen D et al. Use of Autofluorescence to Intraoperatively Diagnose Visceral Pleural Invasion From Frozen Sections in Patients With Lung Adenocarcinoma 2 cm or Less. Am J Clin Pathol. 2019;152(5):608-15. https:// doi.org/10.1093/ajcp/aqz081
[10] Kriegmair MC, Rother J, Grychtol B et al. Multiparametric Cystoscopy for Detection of Bladder Cancer Using Real- time Multispectral Imaging. Eur Urol. 2020;77(2):251-9. https://doi.org/10.1016/j.eururo.2019.08.024
[11] Bae SJ, Lee DS, Berezin V et al. Multispectral autofluorescence imaging for detection of cervical lesions: A preclinical study. J Obstet Gynaecol Res. 2016;42(12):1846-53. https://doi.org/10.1111/jog.13101
[12] McAlpine JN, El Hallani S, Lam SF et al. Autofluorescence imaging can identify preinvasive or clinically occult lesions in fallopian tube epithelium: a promising step towards screening and early detection. Gynecol Oncol. 2011;120(3):385-92. https://doi.org/10.1016/j. ygyno.2010.12.333
[13] Feng PH, Chen TT, Lin YT et al. Classification of lung cancer subtypes based on autofluorescence bronchoscopic pattern recognition: A preliminary study. Comput Methods Programs Biomed. 2018;163:33-8. https://doi. org/10.1016/j.cmpb.2018.05.016
[14] Tamosiunas M, Plorina EV, Lange M et al. Autofluorescence imaging for recurrence detection in skin cancer postoperative scars. J Biophotonics. 2020;13(3):e201900162. https://doi.org/10.1002/ jbio.201900162
[15] Shin EJ, Seo JK, Lee EJ et al. Diagnostic utility of skin autofluorescence when patch test results are doubtful. Skin Res Technol. 2019;25(1):96-9. https://doi.org/10.1111/ srt.12615
[16] Baschong W, Suetterlin R, Laeng RH. Control of autofluorescence of archival formaldehyde-fixed, paraffin- embedded tissue in confocal laser scanning microscopy (CLSM). J Histochem Cytochem. 2001;49(12):1565-72. https://doi.org/10.1177/002215540104901210
17] Erben T, Ossig R, Naim HY et al. What to do with high autofluorescence background in pancreatic tissues - an efficient Sudan black B quenching method for specific immunofluorescence labelling. Histopathology. 2016;69(3):406-22. https://doi.org/10.1111/his.12935
[18] Oliveira VC, Carrara RC, Simoes DL et al. Sudan Black B treatment reduces autofluorescence and improves resolution of in situ hybridization specific fluorescent signals of brain sections. Histol Histopathol. 2010;25(8):1017-24. https://doi.org/10.14670/HH-25.1017
[19] Su W, Yang L, Luo X et al. Elimination of Autofluorescence in Archival Formaldehyde-Fixed, Paraffin-Embedded Bone Marrow Biopsies. Arch Pathol Lab Med. 2019;143(3):362-9. https://doi.org/10.5858/arpa.2017-0480-OA
[20] Jones JD, Ramser HE, Woessner AE et al. Quantifying Age-Related Changes in Skin Wound Metabolism Using In Vivo Multiphoton Microscopy. Adv Wound Care (New Rochelle). 2020;9(3):90-102. https://doi.org/10.1089/ wound.2019.1030
[21] Li H, Yan M, Yu J et al. In vivo identification of arteries and veins using two-photon excitation elastin autofluorescence. J Anat. 2020;236(1):171-9. https://doi. org/10.1111/joa.13080