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Blood vessels are the key parts in the tumor microenvironment, which is taken as a new visible target for tumor therapy. Multiphoton microscopy (MPM), based on the second harmonic generation and two\u2010photon excited fluorescence, is available to make the label\u2010free analysis on the blood vessels in human gliomas. MPM can reveal the vascular morphological characteristics in gliomas, including vascular malformation, intense vascular proliferation, perivascular collagen deposition, perivascular lymphocytes aggregation and microvascular proliferation. In addition, the image analysis algorithms were developed to automatically calculate the perivascular collagen content, vascular cavity area, lumen area, wall area and vessel number. Thus, the vascular morphology, the perivascular collagen deposition and intense vascular proliferation degree can be further quantitatively characterized. Compared with the pathological analysis, the combination of MPM and image analysis has potential advantages in making a quantitative and qualitative analyzing on vascular morphology in glioma microenvironment. As micro\u2010endoscope and two\u2010photon fiberscope are technologically improved, this combined method will be a useful imaging way to make the real\u2010time research on the targeting tumor microenvironment in gliomas.<jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/jbio201900006-gra-0001.png\" xlink:title=\"image\"\/><\/jats:p>","DOI":"10.1002\/jbio.201900006","type":"journal-article","created":{"date-parts":[[2019,3,14]],"date-time":"2019-03-14T04:59:03Z","timestamp":1552539543000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Automatic and label\u2010free identification of blood vessels in gliomas using the combination of multiphoton microscopy and image analysis"],"prefix":"10.1002","volume":"12","author":[{"given":"Na","family":"Fang","sequence":"first","affiliation":[{"name":"Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology Fujian Normal University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zanyi","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xingfu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathology The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dezhi","family":"Kang","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lianhuang","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology Fujian Normal University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yupeng","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pathology The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xianying","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Radiology The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shanshan","family":"Cai","sequence":"additional","affiliation":[{"name":"Department of Pathology The Second Affiliated Hospital of Fujian Medical University  Quanzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xueyong","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pathology The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhida","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology Fujian Normal University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haohua","family":"Tu","sequence":"additional","affiliation":[{"name":"Beckman Institute for Advanced Science and Technology University of Illinois at Urbana\u2010Champaign  Urbana Illinois"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanxiang","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery The First Affiliated Hospital of Fujian Medical University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8519-4462","authenticated-orcid":false,"given":"Jianxin","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology Fujian Normal University  Fuzhou People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2019,5,30]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/neuonc\/now129"},{"issue":"4","key":"e_1_2_8_3_1","first-page":"CD007415","volume":"30","author":"Hart M. 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