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Ex vivo confocal microscopy performs real-time assessment of renal biopsy in non-neoplastic diseases

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Abstract

Background

Ex vivo confocal microscopy is a technique for tissue examination, which generates images of fresh samples with an optical resolution comparable to those obtained by conventional pathology. The objective of this study was to evaluate the feasibility of using ex vivo confocal microscopy in fusion mode (reflectance and fluorescence) and the H&E-like digital staining that is obtained for the analysis of non-neoplastic kidney biopsies.

Methods

Twenty-four renal samples acquired from autopsies were scanned in a 4th generation ex vivo confocal microscopy device. The imaging process was completed in an average of three minutes.

Results

Confocal images correlated very well to the corresponding conventional histological sections, both in normal tissue and in chronic lesions (glomerulosclerosis, fibrosis and tubular atrophy). The ex vivo confocal microscopy protocol did not add artifacts to the sample for the ulterior study with light microscopy, nor to the histochemical or immunohistochemical studies.

Conclusion

The ease and speed of grayscale and fluorescence image acquisition, together with the quality of the H&E-like digitally stained images obtained with this approach, suggest that this technique shows promise for use in clinical nephrology and renal transplantation.

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References

  1. Fiorentino M, Bolignano D, Tesar V et al (2016) Renal biopsy in 2015–from epidemiology to evidence-based indications. Am J Nephrol 43:1–19. https://doi.org/10.1159/000444026

    Article  PubMed  Google Scholar 

  2. D’Agati VD, Cohen DJ (2003) Preimplantation renal biopsy: structure does predict function. Transplantation 75:264–266. https://doi.org/10.1097/01.tp.0000045218.25209.af

    Article  PubMed  Google Scholar 

  3. Welker M-W, Weiler N, Bechstein WO et al (2019) Key role of renal biopsy in management of progressive chronic kidney disease in liver graft recipients. J Nephrol 32:129–137. https://doi.org/10.1007/s40620-018-0506-2

    Article  PubMed  Google Scholar 

  4. Richards NT, Darby S, Howie AJ et al (1994) Knowledge of renal histology alters patient management in over 40% of cases. Nephrol Dial Transpl 9:1255–1259

    CAS  Google Scholar 

  5. Ragazzi M, Longo C, Piana S (2016) Ex Vivo (fluorescence) confocal microscopy in surgical pathology: state of the art. Adv Anat Pathol 23:159–169. https://doi.org/10.1097/PAP.0000000000000114

    Article  CAS  PubMed  Google Scholar 

  6. Gareau DS, Jeon H, Nehal KS, Rajadhyaksha M (2012) Rapid screening of cancer margins in tissue with multimodal confocal microscopy. J Surg Res 178:533–538. https://doi.org/10.1016/j.jss.2012.05.059

    Article  PubMed  PubMed Central  Google Scholar 

  7. Cinotti E, Perrot J-L, Labeille B et al (2015) Handheld reflectance confocal microscopy for the diagnosis of conjunctival tumors. Am J Ophthalmol 159:324–33.e1. https://doi.org/10.1016/j.ajo.2014.10.031

    Article  PubMed  Google Scholar 

  8. Alarcon I, Carrera C, Palou J et al (2014) Impact of in vivo reflectance confocal microscopy on the number needed to treat melanoma in doubtful lesions. Br J Dermatol 170:802–808. https://doi.org/10.1111/bjd.12678

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Combalia M, Pérez-Anker J, García-Herrera A et al (2019) Digitally stained confocal microscopy through deep learning. In: Cardoso MJ, Feragen A, Glocker B, et al. (eds) Proceedings of the 2nd International Conference on medical imaging with deep learning. PMLR, London, pp 121–129

    Google Scholar 

  10. Pérez-Anker J, Ribero S, Yélamos O et al (2020) Basal cell carcinoma characterization using fusion ex vivo confocal microscopy: a promising change in conventional skin histopathology. Br J Dermatol 182:468–476. https://doi.org/10.1111/bjd.18239

    Article  CAS  PubMed  Google Scholar 

  11. Puliatti S, Bertoni L, Pirola GM et al (2019) Ex vivo fluorescence confocal microscopy: the first application for real-time pathological examination of prostatic tissue. BJU Int 124:469–476. https://doi.org/10.1111/bju.14754

    Article  PubMed  Google Scholar 

  12. Krishnamurthy S, Cortes A, Lopez M et al (2018) Ex vivo confocal fluorescence microscopy for rapid evaluation of tissues in surgical pathology practice. Arch Pathol Lab Med 142:396–401. https://doi.org/10.5858/arpa.2017-0164-OA

    Article  PubMed  Google Scholar 

  13. Saco A, Diaz A, Hernandez M et al (2017) Validation of whole-slide imaging in the primary diagnosis of liver biopsies in a University Hospital. Dig liver Dis 49:1240–1246. https://doi.org/10.1016/j.dld.2017.07.002

    Article  PubMed  Google Scholar 

  14. Giataganas P, Hughes M, Payne CJ et al (2019) Intraoperative robotic-assisted large-area high-speed microscopic imaging and intervention. IEEE Trans Biomed Eng 66:208–216. https://doi.org/10.1109/TBME.2018.2837058

    Article  PubMed  Google Scholar 

  15. Yoshitake T, Giacomelli MG, Cahill LC et al (2016) Direct comparison between confocal and multiphoton microscopy for rapid histopathological evaluation of unfixed human breast tissue. J Biomed Opt 21:126021. https://doi.org/10.1117/1.JBO.21.12.126021

    Article  PubMed  PubMed Central  Google Scholar 

  16. Bertoni L, Puliatti S, Reggiani Bonetti L et al (2020) Ex vivo fluorescence confocal microscopy: prostatic and periprostatic tissues atlas and evaluation of the learning curve. Virchows Arch 476:511–520. https://doi.org/10.1007/s00428-019-02738-y

    Article  CAS  PubMed  Google Scholar 

  17. Forest F, Cinotti E, Yvorel V et al (2015) Ex vivo confocal microscopy imaging to identify tumor tissue on freshly removed brain sample. J Neurooncol 124:157–164. https://doi.org/10.1007/s11060-015-1832-z

    Article  PubMed  Google Scholar 

  18. Eschbacher JM, Georges JF, Belykh E et al (2017) Immediate label-free ex vivo evaluation of human brain tumor biopsies with confocal reflectance microscopy. J Neuropathol Exp Neurol 76:1008–1022. https://doi.org/10.1093/jnen/nlx089

    Article  CAS  PubMed  Google Scholar 

  19. Schüürmann M, Stecher MM, Paasch U et al (2019) Evaluation of digital staining for ex vivo confocal laser scanning microscopy. J Eur Acad Dermatol Venereol. https://doi.org/10.1111/jdv.16085

    Article  PubMed  Google Scholar 

  20. Malkoç E, Maurice MJ, Akça O et al (2018) Significance of the nonneoplastic renal parenchymal findings in robotic partial nephrectomy series. J Nephrol 31:925–930. https://doi.org/10.1007/s40620-018-0479-1

    Article  PubMed  Google Scholar 

  21. Girolami I, Gambaro G, Ghimenton C et al (2020) Pre-implantation kidney biopsy: value of the expertise in determining histological score and comparison with the whole organ on a series of discarded kidneys. J Nephrol 33:167–176. https://doi.org/10.1007/s40620-019-00638-7

    Article  PubMed  Google Scholar 

  22. Liapis H, Gaut JP, Klein C et al (2017) Banff histopathological consensus criteria for preimplantation kidney biopsies. Am J Transpl 17:140–150. https://doi.org/10.1111/ajt.13929

    Article  CAS  Google Scholar 

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Acknowledgements

CERCA Program / Generalitat de Catalunya

Funding

This work was supported by: “Instituto de Salud Car-los III (ISCII)” and “Fondo Europeo de Desarrollo Regional (FEDER)”(PI17/00080). Role of digital pathology and nuclear magnetic resonance in the early detection of interstitial fibrosis in chronic kidney disease. "Recent development of the technology within a European Union-funded program (DIAGNOPTICS, " Diagnosis of skin cancer using optics ". Grant agreement no: 621066) enabled to produce a multimodal confocal device for the fast evaluation of tissue specimens Ex-vivo”. Premio Fin de Residencia Hospital Clínic Emili Letang 18_ 251226_PFR_2018. Role of ex vivo confocal microscopy in the evaluation of the renal biopsy.

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Author notes

  1. Jesús Z. Villarreal and J. Pérez-Anker contributed equally to this manuscript.

  2. Luis F. Quintana and A. García-Herrera share senior authorship.

Authors and Affiliations

  1. Nephrology and Renal Transplantation Department, Hospital Clínic de Barcelona, Universitat de Barcelona, Barcelona, Spain

    Jesús Z. Villarreal & Luis F. Quintana

  2. Institut d’Investigacions Biomediques August Pi I Sunyer (IDIBAPS), Barcelona, Spain

    Jesús Z. Villarreal & Luis F. Quintana

  3. Dermatology Department, Melanoma Unit, Hospital Clínic de Barcelona, Institut d’Investigacions Biomediques August Pi I Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain

    J. Pérez-Anker, S. Puig & J. Malvehy

  4. Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Barcelona, Spain

    S. Puig & J. Malvehy

  5. Department of Dermatology, University of Modena and Reggio Emilia, Modena, Italy

    G. Pellacani

  6. Pathology Department, Hospital Clínic de Barcelona, Universitat de Barcelona, Barcelona, Spain

    M. Solé & A. García-Herrera

  7. Centro de Referencia en Enfermedad Glomerular Compleja del Sistema Nacional de Salud (CSUR), Barcelona, Spain

    M. Solé, Luis F. Quintana & A. García-Herrera

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  1. Jesús Z. Villarreal
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  2. J. Pérez-Anker
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Contributions

AGH, LFQ, and JM designed the study; JM, SP and LFQ obtained funding; JZV, JPA and AGH carried out experiments. AGH, MS, GP, JPA, JZV and LFQ analyzed the data; AGH and MS made the figures; AGH, LFQ, JM, MB, JZV, SP, JPA drafted and revised the paper; all authors approved the final version of the manuscript.

Corresponding author

Correspondence to Luis F. Quintana.

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Conflict of interest

No author has any conflict of interest. The information presented in this paper has not been published previously in whole or in part.

Ethical approval

The study was approved by the ethics committee of the Hospital Clinic, Barcelona and was conducted in accordance with the precepts of the Helsinki Declaration. All the data were handled anonymously.

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Villarreal, J.Z., Pérez-Anker, J., Puig, S. et al. Ex vivo confocal microscopy performs real-time assessment of renal biopsy in non-neoplastic diseases. J Nephrol 34, 689–697 (2021). https://doi.org/10.1007/s40620-020-00844-8

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  • DOI: https://doi.org/10.1007/s40620-020-00844-8

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