Confocal laser microscopy without fluorescent dye in minimal-invasive thoracic surgery: an ex-vivo pilot study in lung cancer
-
David Benjamin Ellebrecht
,
Christiane Kuempers
Abstract
Cancer will be the leading cause of death in a few decades. In line with minimal invasive lung cancer surgery, surgeons loose most of their tactile tissue information and need an additional tool of intraoperative tissue navigation during surgery. Confocal laser microscopy is a well-established method of tissue investigation. In this ex-vivo pilot study, we evaluated an endoscopic confocal laser microscope (eCLM) that does not need any fluorescent dye as a diagnostic tool in non-malignant and malignant pulmonary tissue and distal stapler resection margins, respectively. In seven cases, an eCLM was used for examining pulmonary tissue ex-vivo. Images of non-malignant and non-small cell lung cancer tissue and distal stapler resection margins were characterized in terms of specific signal-patterns. No fluorescent dye was used. Correlations to findings in conventional histology were systematically recorded and described. Healthy lung tissue showed hyperreflectoric alveolar walls with dark alveolar spaces. Hyperreflective nets indicated the tumor stroma; whereas the hyperreflective areas indicated the tumor cell clusters. Compared to adenocarcinoma tissue, tissue from squamous cell carcinoma showed more distinctive hyperreflective stroma nets. eCLM characteristics seen in non-malignant and malignant tissue were also visible in distal stapler resection margins and so therefore it was feasible to distinguish between healthy lung tissue and lung cancer. This pilot study shows that the assessment of pulmonary tissue with this eCLM for minimally invasive surgical approach without any fluorescent dye is feasible. It enables to differentiate between benign and malignant tissue in pulmonary specimen by easy to evaluate and reproducible parameters.
Acknowledgments
We would like to thank Prof. Dr. Gereon Hüttmann for valuable discussions.
Research funding: This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Author contributions: Study concept and design: David B. Ellebrecht, Christiane Kuempers, Christian Kugler, and Markus Kleemann. Acquisition of data: David B. Ellebrecht, Christiane Kuempers, and Sven Perner. Analysis and interpretation: David B. Ellebrecht, Christiane Kuempers, Christian Kugler, and Markus Kleemann. Study supervision: David B. Kleemann, and Markus Kleemann. All authors have provided final approval of the version submitted.
Competing interest: The authors declare no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Informed consent: Informed consent was obtained from all individuals included in this study.
Ethical approval: Research involving human subjects complied with all relevant national regulations, data were obtained in accordance with the local ethical committee (NR. 15–160A).
References
1. Dagenais, GR, Leong, DP, Rangarajan, S, Lanas, F, Lopez-Jaramillo, P, Gupta, R, et al.. Variations in common diseases, hospital admissions, and deaths in middle-aged adults in 21 countries from five continents (PURE): a prospective cohort study. Lancet 2020;395:785–94. https://doi.org/10.1016/s0140-6736(19)32007-0.Search in Google Scholar
2. Bray, F, Ferlay, J, Soerjomataram, I, Siegel, RL, Torre, LA, Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018;68:394–424. https://doi.org/10.3322/caac.21492.Search in Google Scholar
3. World Health Organization. The top 10 causes of death; 2018. Available from: https://www.who.int/en/news-room/fact-sheets/detail/the-top-10-causes-of-death.Search in Google Scholar
4. Krebs in Deutschland 2013/2014. Gesellschaft der epidemiologischen Krebsregister in Deutschland e.V. Berlin: Robert Koch-Institut; 2017.Search in Google Scholar
5. Ellebrecht, DB, Latus, S, Schlaefer, A, Keck, T, Gessert, N. Towards an optical biopsy during visceral surgical interventions. Vis Med 2020;36:70–9. https://doi.org/10.1159/000505938.Search in Google Scholar
6. Fottner, C, Mettler, E, Goetz, M, Schirrmacher, E, Anlauf, M, Strand, D, et al.. In vivo molecular imaging of somatostatin receptors in pancreatic islet cells and neuroendocrine tumors by miniaturized confocal laser-scanning fluorescence microscopy. Endocrinology 2010;151:2179–88. https://doi.org/10.1210/en.2009-1313.Search in Google Scholar
7. Fugazza, A, Gaiani, F, Carra, MC, Brunetti, F, Levy, M, Sobhani, I, et al.. Confocal laser endomicroscopy in gastrointestinal and pancreatobiliary diseases: a systematic review and meta-analysis. BioMed Res Int 2016;2016:4638683. https://doi.org/10.1155/2016/4638683.Search in Google Scholar
8. Kiesslich, R, Burg, J, Vieth, M, Gnaendiger, J, Enders, M, Delaney, P, et al.. Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo. Gastroenterology 2004;127:706–13. https://doi.org/10.1053/j.gastro.2004.06.050.Search in Google Scholar
9. Kiesslich, R, Gossner, L, Goetz, M, Dahlmann, A, Vieth, M, Stolte, M, et al.. In vivo histology of Barrett’s esophagus and associated neoplasia by confocal laser endomicroscopy. Clin Gastroenterol Hepatol 2006;4:979–87. https://doi.org/10.1016/j.cgh.2006.05.010.Search in Google Scholar
10. Lim, LG, Neumann, J, Hansen, T, Goetz, M, Hoffman, A, Neurath, MF, et al.. Confocal endomicroscopy identifies loss of local barrier function in the duodenum of patients with Crohn’s disease and ulcerative colitis. Inflamm Bowel Dis 2014;20:892–900. https://doi.org/10.1097/mib.0000000000000027.Search in Google Scholar
11. Streba, CT, Giltan, AM, Gheonea, IA, Demetrian, A, Soimu, AV, Saftoiu, A, et al.. Utility of confocal laser endomicroscopy in pulmonology and lung cancer. Rom J Morphol Embryol 2016;57:1221–7.Search in Google Scholar
12. Ellebrecht, DB, Gebhard, MP, Horn, M, Keck, T, Kleemann, M. Laparoscopic confocal laser microscopy without fluorescent injection: a pilot ex vivo study in colon cancer. Surg Innovat 2016;23:341–6. https://doi.org/10.1177/1553350616637690.Search in Google Scholar
13. Ellebrecht, DB, Kuempers, C, Horn, M, Keck, T, Kleemann, M. Confocal laser microscopy as novel approach for real-time and in-vivo tissue examination during minimal-invasive surgery in colon cancer. Surg Endosc 2019;33:1811–7. https://doi.org/10.1007/s00464-018-6457-9.Search in Google Scholar
14. Stachs, O, Guthoff, RF, Aumann, S. In vivo confocal scanning laser microscopy. In: Bille, JF, editor. High resolution imaging in microscopy and ophthalmology: new frontiers in biomedical optics. Springer International Publishing; 2019. https://doi.org/10.1007/978-3-030-16638-0_12.Search in Google Scholar
15. Thiberville, L, Salaun, M, Lachkar, S, Dominique, S, Moreno-Swirc, S, Vever-Bizet, C, et al.. Human in vivo fluorescence microimaging of the alveolar ducts and sacs during bronchoscopy. Eur Respir J 2009;33:974–85. https://doi.org/10.1183/09031936.00083708.Search in Google Scholar
16. Thiberville, L, Moreno-Swirc, S, Vercauteren, T, Peltier, E, Cave, C, Bourg Heckly, G. In vivo imaging of the bronchial wall microstructure using fibered confocal fluorescence microscopy. Am J Respir Crit Care Med 2007;175:22–31. https://doi.org/10.1164/rccm.200605-684oc.Search in Google Scholar
17. Sorokina, A, Danilevskaya, O, Averyanov, A, Zabozlaev, F, Sazonov, D, Yarmus, L, et al.. Comparative study of ex vivo probe-based confocal laser endomicroscopy and light microscopy in lung cancer diagnostics. Respirology 2014;19:907–13. https://doi.org/10.1111/resp.12326.Search in Google Scholar
18. Fuchs, FS, Zirlik, S, Hildner, K, Schubert, J, Vieth, M, Neurath, MF. Confocal laser endomicroscopy for diagnosing lung cancer in vivo. Eur Respir J 2013;41:1401–8. https://doi.org/10.1183/09031936.00062512.Search in Google Scholar
19. Owen, RM, Force, SD, Gal, AA, Feingold, PL, Pickens, A, Miller, DL, et al.. Routine intraoperative frozen section analysis of bronchial margins is of limited utility in lung cancer resection. Ann Thorac Surg 2013;95:1859–65. https://doi.org/10.1016/j.athoracsur.2012.12.016.Search in Google Scholar
20. Kang, SK, Bok, JS, Cho, HJ, Kang, MW. Novel asymmetrical linear stapler (NALS) for pathologic evaluation of true resection margin tissue. J Thorac Dis 2018;10(14 Suppl):S1631–6. https://doi.org/10.21037/jtd.2018.03.158.Search in Google Scholar
21. Mauna Kea Technologies. Cellvizio® confocal miniprobes. Paris: Mauna Kea Technologies; 2018.Search in Google Scholar
22. Takemura, M, Kurimoto, N, Hoshikawa, M, Maeno, T, Hisada, T, Kurabayashi, M, et al.. Probe-based confocal laser endomicroscopy for rapid on-site evaluation of transbronchial biopsy specimens. Thorac Cancer 2019;10:1441–7. https://doi.org/10.1111/1759-7714.13089.Search in Google Scholar
23. Wellikoff, AS, Holladay, RC, Downie, GH, Chaudoir, CS, Brandi, L, Turbat-Herrera, EA. Comparison of in vivo probe-based confocal laser endomicroscopy with histopathology in lung cancer: a move toward optical biopsy. Respirology 2015;20:967–74. https://doi.org/10.1111/resp.12578.Search in Google Scholar
© 2020 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
- Frontmatter
- Review
- Surrogate based continuous noninvasive blood pressure measurement
- Research Articles
- Smart automated heart health monitoring using photoplethysmography signal classification
- In vivo evaluation of two adaptive Starling-like control algorithms for left ventricular assist devices
- A patient-independent classification system for onset detection of seizures
- Prediction of salivary cortisol level by electroencephalography features
- Confocal laser microscopy without fluorescent dye in minimal-invasive thoracic surgery: an ex-vivo pilot study in lung cancer
- Spinal cord segmentation and injury detection using a Crow Search-Rider optimization algorithm
- Experimental and numerical investigations of fracture and fatigue behaviour of implant-supported bars with distal extension made of three different materials
- Compression and tension behavior of the prosthetic foam materials polyurethane, EVA, Pelite™ and a combination of polyurethane and EVA: a preliminary study
- Evaluation of a novel stair-climbing transportation aid for emergency medical services
Articles in the same Issue
- Frontmatter
- Review
- Surrogate based continuous noninvasive blood pressure measurement
- Research Articles
- Smart automated heart health monitoring using photoplethysmography signal classification
- In vivo evaluation of two adaptive Starling-like control algorithms for left ventricular assist devices
- A patient-independent classification system for onset detection of seizures
- Prediction of salivary cortisol level by electroencephalography features
- Confocal laser microscopy without fluorescent dye in minimal-invasive thoracic surgery: an ex-vivo pilot study in lung cancer
- Spinal cord segmentation and injury detection using a Crow Search-Rider optimization algorithm
- Experimental and numerical investigations of fracture and fatigue behaviour of implant-supported bars with distal extension made of three different materials
- Compression and tension behavior of the prosthetic foam materials polyurethane, EVA, Pelite™ and a combination of polyurethane and EVA: a preliminary study
- Evaluation of a novel stair-climbing transportation aid for emergency medical services
