In current cancer spheroid culturing methods, the transfer and histological processing of specimens grown in 96-well plates is a time consuming process. A centrifugal fluidic device was developed and tested for rapid extraction of spheroids from a 96-well plate and subsequent deposition into a molded agar receiver block. The deposited spheroids must be compact enough to fit into a standard histology cassette while also maintaining a highly planar arrangement. This size and planarity enable histological processing and sectioning of spheroids in a single section. The device attaches directly to a 96-well plate and uses a standard centrifuge to facilitate spheroid transfer. The agar block is then separated from the device and processed. Testing of the device was conducted using six full 96-well plates of fixed Pa14C pancreatic cancer spheroids. On average, 80% of spheroids were successfully transferred into the agar receiver block. Additionally, the planarity of the deposited spheroids was evaluated using confocal laser scanning microscopy. This revealed that, on average, the optimal section plane bisected individual spheroids within 27% of their mean radius. This shows that spheroids are largely deposited in a planar fashion. For rare cases where spheroids had a normalized distance to the plane greater than 1, the section plane either misses or captures a small cross section of the spheroid volume. These results indicate that the proposed device is capable of a high capture success rate and high sample planarity, thus demonstrating the capabilities of the device to facilitate rapid histological evaluation of spheroids grown in standard 96-well plates. Planarity figures are likely to be improved by adjusting agar block handling prior to imaging to minimize deformation and better preserve the planarity of deposited spheroids. Additionally, investigation into media additives to reduce spheroid adhesion to 96-well plates would greatly increase the capture success rate of this device.

中文翻译：

---

用于填充球形癌细胞培养物微阵列的离心流体装置的设计和测试

在目前的癌症球体培养方法中，在 96 孔板中生长的标本的转移和组织学处理是一个耗时的过程。开发并测试了一种离心流体装置，用于从 96 孔板中快速提取球体并随后沉积到模制的琼脂接收器块中。沉积的球体必须足够紧凑以适合标准组织学盒，同时还保持高度平面排列。这种尺寸和平面度可以在单个部分中对球体进行组织学处理和切片。该设备直接连接到 96 孔板并使用标准离心机来促进球体转移。然后将琼脂块与装置分离并进行处理。使用六个完整的 96 孔板固定 Pa14C 胰腺癌球体对该设备进行了测试。平均而言，80% 的球体成功转移到琼脂接收器块中。此外，使用共聚焦激光扫描显微镜评估了沉积球体的平面度。这表明，平均而言，最佳剖面将单个球体平分在其平均半径的 27% 内。这表明球体主要以平面方式沉积。对于球体与平面的归一化距离大于 1 的极少数情况，截面平面要么错过要么捕获球体体积的小横截面。这些结果表明，所提出的装置能够实现高捕获成功率和高样品平面度，从而证明了该装置能够促进在标准 96 孔板中生长的球体的快速组织学评估。通过在成像前调整琼脂块处理以最大限度地减少变形并更好地保持沉积球体的平面度，平面图可能会得到改善。此外，研究介质添加剂以减少球体对 96 孔板的粘附将大大提高该设备的捕获成功率。