Department of Radiation Oncology, Gunma University Graduate School of Medicine, 3-39-22, Showa-machi, Maebashi, Gunma, 371-8511, Japan.
Gunma University Heavy Ion Medical Center, 3-39-22, Showa-machi, Maebashi, Gunma, 371-8511, Japan.
Med Mol Morphol. 2021 Jun;54(2):166-172. doi: 10.1007/s00795-020-00277-z. Epub 2021 Jan 26.
To obtain quantitative volumetric data for the Golgi apparatus after ionizing radiation (IR) using super-resolution three-dimensional structured illumination (3D-SIM) microscopy. Normal human retinal pigment epithelial (RPE) cells were irradiated with X-rays (10 Gy), followed by immunofluorescence staining of the Golgi marker RCAS1. 3D-SIM imaging was performed using DeltaVision OMX version 4 and SoftWoRx 6.1. Polygon rendering and spot signal identification were performed using Imaris 8.1.2. Differences between groups were assessed by Welch's t test. RCAS1 signals in untreated cells were located adjacent to nuclei and showed a reticular morphology. Upon IR, the area of RCAS1 signals expanded while retaining the reticular morphology. Polygon rendering imaging revealed that the volume of RCAS1 at 48 h post-IR was greater than that for unirradiated cells (93.7 ± 19.0 μm vs. 33.0 ± 4.2 μm, respectively; P < 0.001): a 2.8-fold increase. Spot signal imaging showed that the number of RCAS1 spot signals post-IR was greater than that for unirradiated cells [3.4 ± 0.8 (× 10) versus 1.3 ± 0.2 (× 10), respectively; P < 0.001]: a 2.7-fold increase. This is the first study to report quantitative volumetric data of the Golgi apparatus in response to IR using super-resolution 3D-SIM microscopy.
为了使用超分辨率三维结构照明（3D-SIM）显微镜获得电离辐射（IR）后高尔基器的定量体积数据。用 X 射线（10Gy）照射正常人视网膜色素上皮（RPE）细胞，然后用高尔基标志物 RCAS1 进行免疫荧光染色。使用 DeltaVision OMX 版本 4 和 SoftWoRx 6.1 进行 3D-SIM 成像。使用 Imaris 8.1.2 进行多边形渲染和斑点信号识别。通过 Welch's t 检验评估组间差异。未经处理的细胞中 RCAS1 信号位于细胞核附近，呈网状形态。在 IR 后，RCAS1 信号的面积扩大，同时保持网状形态。多边形渲染成像显示，IR 后 48 小时的 RCAS1 体积大于未照射细胞（分别为 93.7±19.0μm 和 33.0±4.2μm，P<0.001）：增加了 2.8 倍。斑点信号成像显示，IR 后 RCAS1 斑点信号的数量大于未照射细胞[3.4±0.8（×10）与 1.3±0.2（×10），P<0.001]：增加了 2.7 倍。这是第一项使用超分辨率 3D-SIM 显微镜报告 IR 后高尔基器定量体积数据的研究。
