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The combination of two‐dimensional and three‐dimensional analysis methods contributes to the understanding of glioblastoma spatial heterogeneity
Author(s) -
Yang Runwei,
Guo Jinglin,
Lin Zhiying,
Song Haimin,
Feng Zhanpeng,
Ou Yichao,
Zhou Mingfeng,
Li Yaomin,
Yi Guozhong,
Li Ke,
Li Kaishu,
Guo Manlan,
Wang Xiran,
Huang Guanglong,
Liu Zhifeng,
Qi Songtao,
Liu Yawei
Publication year - 2020
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201900196
Subject(s) - spatial heterogeneity , glioblastoma , tumour heterogeneity , pathology , spatial distribution , cell , immunohistochemistry , genetic heterogeneity , high resolution , biology , phenotype , medicine , cancer research , genetics , cancer , mathematics , geography , ecology , remote sensing , gene , statistics
Abstract Heterogeneity is regarded as the major factor leading to the poor outcomes of glioblastoma (GBM) patients. However, conventional two‐dimensional (2D) analysis methods, such as immunohistochemistry and immunofluorescence, have limited capacity to reveal GBM spatial heterogeneity. Thus, we sought to develop an effective analysis strategy to increase the understanding of GBM spatial heterogeneity. Here, 2D and three‐dimensional (3D) analysis methods were compared for the examination of cell morphology, cell distribution and large intact structures, and both types of methods were employed to dissect GBM spatial heterogeneity. The results showed that 2D assays showed only cross‐sections of specimens but provided a full view. To visualize intact GBM specimens in 3D without sectioning, the optical tissue clearing methods CUBIC and iDISCO+ were used to clear opaque specimens so that they would become more transparent, after which the specimens were imaged with a two‐photon microscope. The 3D analysis methods showed specimens at a large spatial scale at cell‐level resolution and had overwhelming advantages in comparison to the 2D methods. Furthermore, in 3D, heterogeneity in terms of cell stemness, the microvasculature, and immune cell infiltration within GBM was comprehensively observed and analysed. Overall, we propose that 2D and 3D analysis methods should be combined to provide much greater detail to increase the understanding of GBM spatial heterogeneity.