Tivig Ioan, Moisescu Mihaela Georgeta, Savopol Tudor
Biophysics and Cellular Biotechnology Department, Excellence Center for Research in Biophysics and Cellular Biotechnology, Faculty of Medicine, Carol Davila University of Medicine and Pharmacy, 8 Eroii Sanitari Blvd., Bucharest 050474, Romania.
ACS Omega. 2023 Oct 6;8(41):38715-38722. doi: 10.1021/acsomega.3c06052. eCollection 2023 Oct 17.
Dielectrophoretic (DEP) cell separation, which utilizes electric fields to selectively manipulate and separate cells based on their electrical properties, has emerged as a cutting-edge label-free technique. DEP separation techniques rely on differences in the electrical and morphological properties of cells, which can be obtained by a thorough analysis of DEP spectra. This article presents a novel platform, named OpenDEP, for acquiring and processing DEP spectra of suspended cells. The platform consists of lab-on-a-chip and open-source software that enables the determination of DEP spectra and electric parameters. The performance of OpenDEP was validated by comparing the results obtained using this platform with the results obtained using a commercially available device, 3DEP from DEPtech. The lab-on-a-chip design features two indium tin oxide-coated slides with a specific geometry, forming a chamber where cells are exposed to an inhomogeneous alternating electric field with different frequencies, and microscopic images of cell distributions are acquired. A custom-built software written in the Python programing language was developed to convert the acquired images into DEP spectra, allowing for the estimation of membrane and cytoplasm conductivities and permittivities. The platform was validated using two cell lines, DC3F and NIH 3T3. The OpenDEP platform offers several advantages, including easy manufacturing, statistically robust computations due to large cell population analysis, and a closed environment for sterile work. Furthermore, continuous observation using any microscope allows for integration with other techniques.
介电泳（DEP）细胞分离技术利用电场根据细胞的电学性质对细胞进行选择性操控和分离，已成为一种前沿的无标记技术。DEP分离技术依赖于细胞电学和形态学性质的差异，这些差异可通过对DEP光谱的深入分析获得。本文介绍了一种名为OpenDEP的新型平台，用于获取和处理悬浮细胞的DEP光谱。该平台由芯片实验室和开源软件组成，能够测定DEP光谱和电学参数。通过将使用该平台获得的结果与使用DEPtech公司的商用设备3DEP获得的结果进行比较，验证了OpenDEP的性能。芯片实验室设计的特点是有两个具有特定几何形状的氧化铟锡涂层载玻片，形成一个腔室，细胞在其中暴露于不同频率的非均匀交变电场中，并采集细胞分布的显微图像。开发了一种用Python编程语言编写的定制软件，用于将采集到的图像转换为DEP光谱，从而估计细胞膜和细胞质的电导率和介电常数。该平台使用两种细胞系DC3F和NIH 3T3进行了验证。OpenDEP平台具有多个优点，包括易于制造、由于对大量细胞群体进行分析而具有统计稳健性的计算，以及用于无菌操作的封闭环境。此外，使用任何显微镜进行连续观察可实现与其他技术的整合。
