Year: 2022
Type: article
Abstract: Nanomaterials are the subject of a range of biomedical, commercial, and environmental investigations involving measurements in living cells and tissues. Accurate quantification of nanomaterials, at the tissue, cell, and organelle levels, is often difficult, however, in part due to their inhomogeneity. Here, we propose a method that uses the diverse optical properties of a nanomaterial preparation in order to improve quantification at the single-cell and organelle level. We developed ‘hyperspectral counting’, which employs diffraction-limited imaging via hyperspectral microscopy of a diverse set of nanomaterial emitters, to estimate nanomaterial counts in live cells and sub-cellular structures. A mathematical model was developed, and Monte Carlo simulations were employed, to improve the accuracy of these estimates, enabling quantification with single-cell and single-endosome resolution. We applied this nanometrology technique to identify an upper-limit of the rate of uptake into cells - approximately 3,000 particles endocytosed within 30 minutes. In contrast, conventional ROI counting results in a 230% undercount. The method identified significant heterogeneity and a broad non-Gaussian distribution of carbon nanotube uptake within cells. For example, while a particular cell contained an average of 1 nanotube per endosome, the heterogenous distribution resulted in over 7 nanotubes localizing within some endosomes, substantially changing the accounting of subcellular nanoparticle concentration distributions. This work presents a method to quantify cellular and subcellular concentrations of a heterogeneous carbon nanotube reference material, with implications for nanotoxicology, drug/gene delivery, and nanosensor fields.
Source: ACS Nano
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
Funders: National Science Foundation, American Cancer Society, Commonwealth Foundation, Honorable Tina Brozman Foundation, Memorial Sloan-Kettering Cancer Center, University of Rhode Island, Rhode Island College, Pershing Square Sohn Cancer Research Alliance, Commonwealth Foundation for Cancer Research Foundation, Cycle for Survival, National Institutes of Health, National Cancer Institute, Division of Chemical, Bioengineering, Environmental, and Transport Systems, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1676388581, G2051006376, G2295061683, G289723095, Understanding and Controlling the Cellular Forces that Coordinate Epithelial Tissue Morphogenesis, G372934033, G4102770861, G4583515593, Transient Metabolite Detection for Single-Cell Metabolomics and Diagnostics, CAREER: Spectral imaging for sub-cellular nanometrology and nanotoxicology, G5109122429, CAREER: A Quantitative Nanosensor to Measure Redox Potential in Living Systems, P-selectin-Mediated Targeting of PI3K Nanomedicines to the Tumor Microenvironment, G6671297155, G6680673728, G6743818077, G675646491, G6812410181, G7556015768, G8525765020, G9000003249, G9000270641, Cancer Center Support Grant
OpenAlex ID: W4220854855