Application of 3D Reflectance Confocal Microscopy: Melanocytic Proliferations as 3D Models; JAAD Supplemental Material

Melanoma on chronically sun-damaged skin (MSDS) is challenging to detect. Reflectance confocal microscopy (RCM) increases diagnostic accuracy, but relies upon interpretation of 2D horizontal sections. Our objective was to determine whether existing technology could be used to create 3D models, which would expand on data available for discriminating skin biology. Supplemental Document: Expanded manuscript describing the background and process. Supplemental Figure 1. Object files. 3D Model from confocal image stack of a selected large atypical cell showing multiple dendrites extending from the body of the cell and nuclear abnormalities. The opacity of the cell body is reduced, such that the nucleus can be seen internally. The object can be rotated freely and examined from any angle using a 3D viewer. NOTE: In order to view the 3D object file interactively and with transparency using Windows 10 3D Viewer, please unzip file containing the .mtl and .obj to the DESKTOP (not a network drive or server). Once both files are on the desktop, double clicking on the .obj file will allow the native 3D viewer to open the model. Supplemental Figure 2. 3D reconstruction of fields of melanoma cells sampled from confocal stacks. The left panel reveals marked pleomorphism while the right panel reveals a relatively uniform, although atypical, collection of cells. Scale bar is 50µm. Supplemental Figure 3. Volume rendering of a stack of confocal images of melanoma on sun-damaged skin viewed along the z axis. Most refractile melanocytic cells are located in the basal layer at approx. 38µm from stratum corneum (yellow arrow), while rare pagetoid cells are present higher in the epidermis (green arrow). This image also reveals marked flattening of the DEJ (orange bracket). Scale Bar is 50µm. Supplemental Figure 4. Movie (.AVI) of a 3D reconstruction of a confocal image stack examining the DEJ by showing the dermal papillae, the blood vessels inside the DP and an adjacent hair follicle.

慢性日光损伤皮肤黑色素瘤（Melanoma on chronically sun-damaged skin, MSDS）的诊断颇具挑战性。反射共聚焦显微镜（Reflectance Confocal Microscopy, RCM）可提升诊断准确性，但依赖对二维水平断层切片的判读。本研究旨在明确能否利用现有技术构建三维模型，以拓展用于区分皮肤生物学特征的可用数据。
补充文档：详述研究背景与实验流程的扩展手稿。
补充图1：对象文件。基于选定的大型非典型细胞共聚焦图像栈构建的三维模型，该细胞可见多个自胞体伸出的树突及核形态异常。胞体透明度降低，可观察到内部的细胞核。该模型可通过三维查看器自由旋转，并从任意角度进行检视。
注意事项：若需在Windows 10的3D查看器中交互式查看该三维对象文件并启用透明效果，请将包含.mtl与.obj格式文件的压缩包解压至桌面（请勿解压至网络驱动器或服务器）。待两个文件均位于桌面后，双击.obj文件即可调用原生3D查看器打开该模型。
补充图2：从共聚焦图像栈中提取的黑色素瘤细胞区域三维重建结果。左侧面板可见显著的细胞多形性，右侧面板则呈现相对均一但仍属非典型的细胞集合。比例尺为50μm。
补充图3：沿Z轴检视的日光损伤皮肤黑色素瘤共聚焦图像栈体积渲染结果。多数折光性黑素细胞位于基底层，距角质层约38μm（黄色箭头），而罕见的佩吉特样细胞则位于表皮较上层（绿色箭头）。该图像同时可见真皮表皮交界处（dermal epidermal junction, DEJ）显著扁平化（橙色括号标注）。比例尺为50μm。
补充图4：共聚焦图像栈的三维重建动画（.AVI格式），通过展示真皮乳头（dermal papilla, DP）、乳头内血管及邻近毛囊来检视真皮表皮交界处。