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01-05-2018 | Original Article

Label-free imaging for T staging of gastric carcinoma by multiphoton microscopy

Authors: Kexin He, Lili Zhao, Xiaoyang Huang, Ying Ding, Li Liu, Xiang Wang, Min Wang, Yong Zhang, Zhining Fan

Published in: Lasers in Medical Science | Issue 4/2018

Abstract

Gastric cancer is one of the most common malignancies worldwide. The accurate diagnosis of tumor invasion depth is critical for therapeutic strategy and prognosis. Without fluorescent labelling, multiphoton microscopy (MPM) imaging could directly reveal tissue architecture based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). In this study, we aimed to explore the feasibility of MPM imaging to assess the gastric tumor morphology and infiltration. Unstained slides of 18 fresh gastric tissues with different T staging were examined by multiphoton microscopy. Morphological and quantitative analyses were both conducted. The nuclear area was defined as the area of nuclear boundary. Collagen content was defined as the ratio of SHG pixels to all pixels. Gastric normal and tumor tissues under different T stages visually presented with cellular and subcellular features on fluorescent imaging. The nuclear areas of normal and cancerous cells were 32.01 ± 2.89 and 58.41 ± 6.06 μm² (P < 0.001), respectively. Collagen content was quantified as 0.087 ± 0.012 in normal mucosa but 0.020 ± 0.007 in cancerous mucosa (P < 0.001). All results were in accord with the paired H&E-stained slides. Our findings suggested the convincing potential of MPM for judging T staging of gastric cancer. Without staining intervention, TPEF and SHG of MPM imaging could objectively and quantitatively indicate the subcellular and molecular changes during carcinogenesis. With the advancement of deep penetration, self-focus imaging and three-dimensional (3D) visualization, label-free MPM imaging compacted with endoscopy could be further introduced to realize the real-time in vivo assessment of tumor invasion clinically.

Siegel RL, Miller KD, Jemal A (2017) Cancer statistics, 2017. CA Cancer J Clin 67(1):7–30. https://doi.org/10.3322/caac.21387 CrossRefPubMed

Van Cutsem E, Sagaert X, Topal B, Haustermans K, Prenen H (2016) Gastric cancer. Lancet (London, England) 388(10060):2654–2664. https://doi.org/10.1016/s0140-6736(16)30354-3 CrossRef

Rausei S, Ruspi L, Galli F, Pappalardo V, Di Rocco G, Martignoni F, Frattini F, Rovera F, Boni L, Dionigi G (2016) Seventh tumor-node-metastasis staging of gastric cancer: five-year follow-up. World J Gastroenterol 22(34):7748–7753. https://doi.org/10.3748/wjg.v22.i34.7748 CrossRefPubMedPubMedCentral

Shen L, Huang Y, Sun M, Xu H, Wei W, Wu W (2009) Clinicopathological features associated with lymph node metastasis in early gastric cancer: analysis of a single-institution experience in China. Can J Gastroenterol = Journal canadien de gastroenterologie 23(5):353–356CrossRefPubMed

Wang Z, Ma L, Zhang XM, Zhou ZX (2014) Risk of lymph node metastases from early gastric cancer in relation to depth of invasion: experience in a single institution. Asian Pac J Cancer Prevention : APJCP 15(13):5371–5375CrossRef

Ma M, Chen S, Zhu BY, Zhao BW, Wang HS, Xiang J, Wu XB, Lin YJ, Zhou ZW, Peng JS, Chen YB (2015) The clinical significance and risk factors of solitary lymph node metastasis in gastric cancer. PLoS One 10(1):e0114939. https://doi.org/10.1371/journal.pone.0114939 CrossRefPubMedPubMedCentral

Xi H, Zhang K, Wei B, Chen L (2017) Significance and contemplation of clinical diagnosis and therapy on the renewal of the eighth edition of gastric cancer TNM staging system. Zhonghua wei chang wai ke za zhi = Chinese journal of gastrointestinal surgery 20(2):166–170PubMed

Association JGC (2017) Japanese gastric cancer treatment guidelines 2014 (ver. 4). Gastric Cancer: Off J Int Gastric Cancer Assoc Jpn Gastric Cancer Assoc 20(1):1–19. https://doi.org/10.1007/s10120-016-0622-4 CrossRef

Arslan H, Fatih Ozbay M, Calli I, Dogan E, Celik S, Batur A, Bora A, Yavuz A, Bulut MD, Ozgokce M, Cetin Kotan M (2017) Contribution of diffusion weighted MRI to diagnosis and staging in gastric tumors and comparison with multi-detector computed tomography. Radiol Oncol 51(1):23–29. https://doi.org/10.1515/raon-2017-0002 CrossRefPubMedPubMedCentral

10.

Han C, Lin R, Shi H, Liu J, Qian W, Ding Z, Hou X (2016) The role of endoscopic ultrasound on the preoperative T staging of gastric cancer: a retrospective study. Medicine 95(36):e4580. https://doi.org/10.1097/md.0000000000004580 CrossRefPubMedPubMedCentral

11.

Hu YF, Deng ZW, Liu H, Mou TY, Chen T, Lu X, Wang D, Yu J, Li GX (2016) Staging laparoscopy improves treatment decision-making for advanced gastric cancer. World J Gastroenterol 22(5):1859–1868. https://doi.org/10.3748/wjg.v22.i5.1859 CrossRefPubMedPubMedCentral

12.

Mehmedovic A, Mesihovic R, Saray A, Vanis N (2014) Gastric cancer staging: EUS and CT. Med Arch (Sarajevo, Bosnia and Herzegovina) 68(1):34–36

13.

Wu CX, Zhu ZH (2014) Diagnosis and evaluation of gastric cancer by positron emission tomography. World J Gastroenterol 20(16):4574–4585. https://doi.org/10.3748/wjg.v20.i16.4574 CrossRefPubMedPubMedCentral

14.

Yan J, Chen G, Chen J, Liu N, Zhuo S, Yu H, Ying M (2011) A pilot study of using multiphoton microscopy to diagnose gastric cancer. Surg Endosc 25(5):1425–1430. https://doi.org/10.1007/s00464-010-1409-z CrossRefPubMed

15.

Zhou Y, Kang D, Yang Z, Li L, Zhuo S, Zhu X, Zhou Y, Chen J (2016) Imaging normal and cancerous human gastric muscular layer in transverse and longitudinal sections by multiphoton microscopy. Scanning 38(4):357–364. https://doi.org/10.1002/sca.21278 CrossRefPubMed

16.

Ma L, Huang X, Wang X, Zhang Y, Liu L, Sheng Y, Fan Z (2015) Study on the diagnosis of rabbit VX2 esophageal cancer and stent-therapy efficacy based on multiphoton microscopy. Scanning 37(2):152–157. https://doi.org/10.1002/sca.21192 CrossRefPubMed

17.

Pimentel-Nunes P, Dinis-Ribeiro M, Ponchon T, Repici A, Vieth M, De Ceglie A, Amato A, Berr F, Bhandari P, Bialek A, Conio M, Haringsma J, Langner C, Meisner S, Messmann H, Morino M, Neuhaus H, Piessevaux H, Rugge M, Saunders BP, Robaszkiewicz M, Seewald S, Kashin S, Dumonceau JM, Hassan C, Deprez PH (2015) Endoscopic submucosal dissection: European Society of Gastrointestinal Endoscopy (ESGE) guideline. Endoscopy 47(9):829–854. https://doi.org/10.1055/s-0034-1392882 CrossRefPubMed

18.

Roukos DH (2000) Current status and future perspectives in gastric cancer management. Cancer Treat Rev 26(4):243–255. https://doi.org/10.1053/ctrv.2000.0164 CrossRefPubMed

19.

Hwang SW, Lee DH (2014) Is endoscopic ultrasonography still the modality of choice in preoperative staging of gastric cancer? World J Gastroenterol 20(38):13775–13782. https://doi.org/10.3748/wjg.v20.i38.13775 CrossRefPubMedPubMedCentral

20.

Mocellin S, Pasquali S (2015) Diagnostic accuracy of endoscopic ultrasonography (EUS) for the preoperative locoregional staging of primary gastric cancer. Cochrane Database Syst Rev (2):Cd009944. doi:https://doi.org/10.1002/14651858.CD009944.pub2

21.

Yoon H, Lee DH (2014) New approaches to gastric cancer staging: beyond endoscopic ultrasound, computed tomography and positron emission tomography. World J Gastroenterol 20(38):13783–13790. https://doi.org/10.3748/wjg.v20.i38.13783 CrossRefPubMedPubMedCentral

22.

Giganti F, Orsenigo E, Arcidiacono PG, Nicoletti R, Albarello L, Ambrosi A, Salerno A, Esposito A, Petrone MC, Chiari D, Staudacher C, Del Maschio A, De Cobelli F (2016) Preoperative locoregional staging of gastric cancer: is there a place for magnetic resonance imaging? Prospective comparison with EUS and multidetector computed tomography. Gastric Cancer: Off J Int Gastric Cancer Assoc Jpn Gastric Cancer Assoc 19(1):216–225. https://doi.org/10.1007/s10120-015-0468-1 CrossRef

23.

Li L, Li H, Chen Z, Zhuo S, Feng C, Yang Y, Guan G, Chen J (2015) Layer-resolved colorectal tissues using nonlinear microscopy. Lasers Med Sci 30(5):1589–1597. https://doi.org/10.1007/s10103-015-1767-9 CrossRefPubMed

24.

Cho HJ, Chun HJ, Kim ES, Cho BR (2011) Multiphoton microscopy: an introduction to gastroenterologists. World J Gastroenterol 17(40):4456–4460. https://doi.org/10.3748/wjg.v17.i40.4456 CrossRefPubMedPubMedCentral

25.

Okada T, Takahashi S, Ishida A, Ishigame H (2016) In vivo multiphoton imaging of immune cell dynamics. Pflugers Archiv : Eur J Physiol 468(11–12):1793–1801. https://doi.org/10.1007/s00424-016-1882-x CrossRef

26.

Malide D (2016) In vivo cell tracking using two-photon microscopy. Methods Mol Biol (Clifton, NJ) 1444:109–122. https://doi.org/10.1007/978-1-4939-3721-9_11 CrossRef

27.

Schultz SR, Copeland CS, Foust AJ, Quicke P, Schuck R (2017) Advances in two photon scanning and scanless microscopy technologies for functional neural circuit imaging. Proceedings of the IEEE Institute of Electrical and Electronics Engineers 105(1):139–157. https://doi.org/10.1109/jproc.2016.2577380 CrossRefPubMed

28.

Wang BG, Konig K, Halbhuber KJ (2010) Two-photon microscopy of deep intravital tissues and its merits in clinical research. J Microsc 238(1):1–20. https://doi.org/10.1111/j.1365-2818.2009.03330.x CrossRefPubMed

29.

Wu Z, Rademakers T, Kiessling F, Vogt M, Westein E, Weber C, Megens RTA, van Zandvoort M (2017) Multi-photon microscopy in cardiovascular research. Methods (San Diego, Calif). doi:https://doi.org/10.1016/j.ymeth.2017.04.013

30.

Wang ZN, Xu HM (2000) Relationship between collagen IV expression and biological behavior of gastric cancer. World J Gastroenterol 6(3):438–439CrossRefPubMedPubMedCentral

31.

Yin Y, Zhao Y, Li AQ, Si JM (2009) Collagen: a possible prediction mark for gastric cancer. Med Hypotheses 72(2):163–165. https://doi.org/10.1016/j.mehy.2008.09.010 CrossRefPubMed

32.

Chen J, Zhuo S, Chen G, Yan J, Yang H, Liu N, Zheng L, Jiang X, Xie S (2011) Establishing diagnostic features for identifying the mucosa and submucosa of normal and cancerous gastric tissues by multiphoton microscopy. Gastrointest Endosc 73(4):802–807. https://doi.org/10.1016/j.gie.2010.12.016 CrossRefPubMed

33.

Li A, Zhou T, Guo L, Si J (2010) Collagen type I regulates beta-catenin tyrosine phosphorylation and nuclear translocation to promote migration and proliferation of gastric carcinoma cells. Oncol Rep 23(5):1247–1255PubMed

34.

Golaraei A, Kontenis L, Cisek R, Tokarz D, Done SJ, Wilson BC, Barzda V (2016) Changes of collagen ultrastructure in breast cancer tissue determined by second-harmonic generation double Stokes-Mueller polarimetric microscopy. Biomed Optics Express 7(10):4054–4068. https://doi.org/10.1364/BOE.7.004054 CrossRef

35.

Matsui H, Kubochi K, Okazaki I, Yoshino K, Ishibiki K, Kitajima M (1999) Collagen biosynthesis in gastric cancer: immunohistochemical analysis of prolyl 4-hydroxylase. J Surg Oncol 70(4):239–246CrossRefPubMed

36.

Makale M, McElroy M, O'Brien P, Hoffman RM, Guo S, Bouvet M, Barnes L, Ingulli E, Cheresh D (2009) Extended-working-distance multiphoton micromanipulation microscope for deep-penetration imaging in live mice and tissue. J Biomed Opt 14(2):024032. https://doi.org/10.1117/1.3103783 CrossRefPubMedPubMedCentral

37.

Liu W, Chia SH, Chung HY, Greinert R, Kartner FX, Chang G (2017) Energetic ultrafast fiber laser sources tunable in 1030-1215 nm for deep tissue multi-photon microscopy. Opt Express 25(6):6822–6831. https://doi.org/10.1364/oe.25.006822 CrossRefPubMed

38.

Cicchi R, Sampson D, Massi D, Pavone F (2005) Contrast and depth enhancement in two-photon microscopy of human skin ex vivo by use of optical clearing agents. Opt Express 13(7):2337–2344CrossRefPubMed

39.

McConnell G (2006) Improving the penetration depth in multiphoton excitation laser scanning microscopy. J Biomed Opt 11(5):054020. https://doi.org/10.1117/1.2360593 CrossRefPubMed

40.

Duan X, Li H, Qiu Z, Joshi BP, Pant A, Smith A, Kurabayashi K, Oldham KR, Wang TD (2015) MEMS-based multiphoton endomicroscope for repetitive imaging of mouse colon. Biomed Optics Express 6(8):3074–3083. https://doi.org/10.1364/boe.6.003074 CrossRef

41.

Rivera DR, Brown CM, Ouzounov DG, Pavlova I, Kobat D, Webb WW, Xu C (2011) Compact and flexible raster scanning multiphoton endoscope capable of imaging unstained tissue. Proc Natl Acad Sci U S A 108(43):17598–17603. https://doi.org/10.1073/pnas.1114746108 CrossRefPubMedPubMedCentral

42.

Brown CM, Rivera DR, Pavlova I, Ouzounov DG, Williams WO, Mohanan S, Webb WW, Xu C (2012) In vivo imaging of unstained tissues using a compact and flexible multiphoton microendoscope. J Biomed Opt 17(4):040505. https://doi.org/10.1117/1.jbo.17.4.040505 CrossRefPubMedPubMedCentral

Title: Label-free imaging for T staging of gastric carcinoma by multiphoton microscopy
Authors: Kexin He
Lili Zhao
Xiaoyang Huang
Ying Ding
Li Liu
Xiang Wang
Min Wang
Yong Zhang
Zhining Fan
Publication date: 01-05-2018
Publisher: Springer London
Published in: Lasers in Medical Science / Issue 4/2018
Print ISSN: 0268-8921
Electronic ISSN: 1435-604X
DOI: https://doi.org/10.1007/s10103-018-2442-8

At a glance: The STEP trials

Springer Medicine

Label-free imaging for T staging of gastric carcinoma by multiphoton microscopy

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

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