Central Laboratory, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Department of Human Reproductive Medicine, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Cytometry A. 2021 Nov;99(11):1114-1122. doi: 10.1002/cyto.a.24358. Epub 2021 May 11.
As label-free biomarkers, electrical properties of single cells have been widely used for cell-type classification and cell-status evaluation. However, as intrinsic bioelectrical markers, previously reported membrane capacitance and cytoplasmic resistance (e.g., specific membrane capacitance C and cytoplasmic conductivity σ ) of tumor subtypes were derived from tens of single cells, lacking statistical significance due to low cell numbers. In this study, tumor subtypes were constructed based on phenotype (treatment with 4-methylumbelliferone) or genotype (knockdown of ROCK1) modifications and then aspirated through a constriction-channel based impedance flow cytometry to characterize single-cell C and σ . Thousands of single tumor cells with phenotype modifications were measured, resulting in significant differences in 1.64 ± 0.43 μF/cm vs. 1.55 ± 0.47 μF/cm of C and 0.96 ± 0.37 S/m vs. 1.24 ± 0.47 S/m of σ for 95C cells (792 cells of 95C-control vs. 1529 cells of 95C-pheno-mod); 2.56 ± 0.88 μF/cm vs. 2.33 ± 0.56 μF/cm of C and 0.83 ± 0.18 S/m vs. 0.93 ± 0.25 S/m of σ for H1299 cells (962 cells of H1299-control vs. 637 cells of H1299-pheno-mod). Furthermore, thousands of single tumor cells with genotype modifications were measured, resulting in significant differences in 3.82 ± 0.92 vs. 3.18 ± 0.47 μF/cm of C and 0.47 ± 0.05 vs. 0.52 ± 0.05 S/m of σ (1100 cells of A549-control vs. 1100 cells of A549-geno-mod). These results indicate that as intrinsic bioelectrical markers, specific membrane capacitance and cytoplasmic conductivity can be used to classify tumor subtypes.
作为无标记生物标志物，单个细胞的电学特性已被广泛用于细胞类型分类和细胞状态评估。然而，作为内在生物电学标记物，先前报道的肿瘤亚型的膜电容和细胞质电阻（例如，特定膜电容 C 和细胞质电导率 σ）是从数十个单个细胞中得出的，由于细胞数量少，缺乏统计学意义。在这项研究中，通过表型（用 4-甲基伞形酮处理）或基因型（敲低 ROCK1）修饰构建肿瘤亚型，然后通过基于收缩通道的阻抗流式细胞术抽吸来表征单个细胞的 C 和 σ。测量了数千个具有表型修饰的单个肿瘤细胞，导致 95C 细胞的 C 值有显著差异，为 1.64 ± 0.43 μF/cm 对 1.55 ± 0.47 μF/cm，σ 值有显著差异，为 0.96 ± 0.37 S/m 对 1.24 ± 0.47 S/m；对于 95C 细胞（95C-对照 792 个细胞对 95C-表型修饰 1529 个细胞）；对于 H1299 细胞，C 值有显著差异，为 2.56 ± 0.88 μF/cm 对 2.33 ± 0.56 μF/cm，σ 值有显著差异，为 0.83 ± 0.18 S/m 对 0.93 ± 0.25 S/m；对于 H1299 细胞（H1299-对照 962 个细胞对 H1299-表型修饰 637 个细胞）。此外，还测量了数千个具有基因型修饰的单个肿瘤细胞，导致 C 值有显著差异，为 3.82 ± 0.92 μF/cm 对 3.18 ± 0.47 μF/cm，σ 值有显著差异，为 0.47 ± 0.05 S/m 对 0.52 ± 0.05 S/m（A549-对照 1100 个细胞对 A549-基因型修饰 1100 个细胞）。这些结果表明，作为内在生物电学标记物，特定膜电容和细胞质电导率可用于肿瘤亚型分类。
