3D printed imaging platform for portable cell counting
3D printed imaging platform for portable cell counting
复制标题
DOI：
10.1039/d1an00778e
复制
发表时间：
2021-05-24
期刊：
影响因子：
4.2
通讯作者：
Juarez，Jaime J.
中科院分区：
文献类型：
--
作者：
Awate，Diwakar M.;Pola，Cicero C.;Juarez，Jaime J.
尽管在生物医学科学中有广泛的应用，但流式细胞仪有一些局限性，阻碍了它们在资源有限的环境中用于即时诊断。3D打印提供了一种经济有效的方法来提高POC设备在资源有限的环境中的可访问性。为了实现这一目标，我们引入了一种3d打印成像平台（3DPIP），能够准确计数颗粒并进行荧光显微镜。在我们的3DPIP中，捕获的颗粒流微观图像在定制开发的颗粒计数器代码上进行处理，以提供颗粒计数。该原型使用基于机器视觉的算法从捕获的流图像中识别颗粒，并且足够灵活，可以进行标记和无标记的颗粒计数。此外，粒子计数器代码返回相对于时间的粒子坐标，该坐标可进一步用于执行粒子图像测速。这些结果可以帮助估计作用在粒子上的力，并识别和分类不同类型的细胞/粒子。我们通过对不同浓度的去离子水稀释后的10 μm聚苯乙烯颗粒进行计数，并将结果与商用Beckman-Coulter Z2颗粒计数器进行比较，评估了该原型的性能。3DPIP可以将颗粒浓度计数到每mL约100个颗粒，标准偏差为±20个颗粒，这与商用颗粒计数器上获得的结果相当。我们的平台在高达9 mL h - 1的流速下产生准确的结果，浓度低于每mL 1000个颗粒，而5 mL h - 1产生准确的结果高于该浓度限制。除了进行流动实验外，我们的仪器还能够进行与平板阅读器相当的静态实验。在这种配置下，我们的仪器能够根据制备的细菌样品（Citrobacter freundii; ATCC 8090）的浓度，每张图像计数10到250个细胞。总的来说，该平台代表了在资源有限的临床环境中开发可负担得起的全3D打印成像流式细胞仪的第一步。
Despite having widespread application in the biomedical sciences, flow cytometers have several limitations that prevent their application to point-of-care (POC) diagnostics in resource-limited environments. 3D printing provides a cost-effective approach to improve the accessibility of POC devices in resource-limited environments. Towards this goal, we introduce a 3D-printed imaging platform (3DPIP) capable of accurately counting particles and perform fluorescence microscopy. In our 3DPIP, captured microscopic images of particle flow are processed on a custom developed particle counter code to provide a particle count. This prototype uses a machine vision-based algorithm to identify particles from captured flow images and is flexible enough to allow for labeled and label-free particle counting. Additionally, the particle counter code returns particle coordinates with respect to time which can further be used to perform particle image velocimetry. These results can help estimate forces acting on particles, and identify and sort different types of cells/particles. We evaluated the performance of this prototype by counting 10 μm polystyrene particles diluted in deionized water at different concentrations and comparing the results with a commercial Beckman-Coulter Z2 particle counter. The 3DPIP can count particle concentrations down to ∼100 particles per mL with a standard deviation of ±20 particles, which is comparable to the results obtained on a commercial particle counter. Our platform produces accurate results at flow rates up to 9 mL h−1 for concentrations below 1000 particle per mL, while 5 mL h−1 produces accurate results above this concentration limit. Aside from performing flow-through experiments, our instrument is capable of performing static experiments that are comparable to a plate reader. In this configuration, our instrument is able to count between 10 and 250 cells per image, depending on the prepared concentration of bacteria samples (Citrobacter freundii; ATCC 8090). Overall, this platform represents a first step towards the development of an affordable fully 3D printable imaging flow cytometry instrument for use in resource-limited clinical environments.