Shin Hye Ji, Kwak Minjeong, Kwon Ik Hwan, Kim Sook Heun, Lee Ji Youn
Biometrology Group, Division of Biomedical Metrology, Korea Research Institute of Standards and Science 267 Gajeong-ro, Yuseong-gu Daejeon 34113 Republic of Korea
Graduate School of Analytical Science and Technology, Chungnam National University 99 Daehak-ro, Yuseong-gu Daejeon 34134 Republic of Korea.
Nanoscale Adv. 2025 Apr 16. doi: 10.1039/d4na00918e.
Quantifying cellular uptake of nanoparticles is critical for understanding their biological interactions and optimizing their applications in nanomedicine. In this study, we developed a flow cytometry-based method to quantify the uptake of gold nanoparticles (AuNPs) using A549 cells. Taking advantage of the scattering properties of AuNPs, this method uses side scatter intensity to estimate the number of nanoparticles internalized by cells. However, directly measuring the exact number of internalized nanoparticles remains challenging due to the tendency of AuNPs to aggregate within cells. To address this, we introduce a new unit, molecules of equivalent gold nanoparticle (MEAuNP), which expresses side scatter intensity as a standardized unit based on the scattering of a single AuNP. While this method does not directly solve the problem of accurately measuring the exact number of internalized nanoparticles, it provides a semi-quantitative approach for estimating nanoparticle uptake. The obtained MEAuNP values are consistent with literature reports, suggesting that the approach yields reliable and comparable data. Moreover, the use of calibrated values ensures that consistent results can be obtained across different acquisition settings and potentially across different instruments. We further examined uptake dynamics and validated the method across multiple cell lines including HeLa, Beas-2B, Jurkat, and RPMI8226. This approach provides a robust tool for quantifying metal nanoparticle uptake, supporting the standardization of estimating uptake levels in various biological systems.
量化纳米颗粒的细胞摄取对于理解它们的生物相互作用以及优化它们在纳米医学中的应用至关重要。在本研究中，我们开发了一种基于流式细胞术的方法，使用A549细胞来量化金纳米颗粒（AuNPs）的摄取。利用AuNPs的散射特性，该方法使用侧向散射强度来估计细胞内化的纳米颗粒数量。然而，由于AuNPs在细胞内聚集的倾向，直接测量内化纳米颗粒的确切数量仍然具有挑战性。为了解决这个问题，我们引入了一个新的单位，等效金纳米颗粒分子（MEAuNP），它基于单个AuNP的散射将侧向散射强度表示为一个标准化单位。虽然这种方法不能直接解决准确测量内化纳米颗粒确切数量的问题，但它提供了一种半定量方法来估计纳米颗粒摄取。获得的MEAuNP值与文献报道一致，表明该方法产生可靠且可比的数据。此外，使用校准值可确保在不同的采集设置下甚至可能在不同的仪器上获得一致的结果。我们进一步研究了摄取动力学，并在包括HeLa、Beas-2B、Jurkat和RPMI8226在内的多种细胞系中验证了该方法。这种方法为量化金属纳米颗粒摄取提供了一个强大的工具，支持在各种生物系统中估计摄取水平的标准化。
